OTAR3088/BioData-CL_Article-Sentences · Datasets at Hugging Face

PMCID string	Title string	Sentences string
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	After overnight incubation, the cells were treated with imatinib at 0, 20, 40, 60, and 80 μmol/L for 48 h. CompuSyn software was used to calculate the combination index using the Chou-Talalay method to determine the antagonistic influence.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	For the migration assay, GIST cells were seeded into 8 µm well Transwell chambers (Corning; Corning, NY, United States).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The upper chamber was filled with 200 µL serum-free medium containing 2 × 10 cells and the lower chamber was filled with 500 μL complete medium (10% FBS).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	After 48 h, the cells were fixed with formaldehyde and stained with 0.2% crystal violet for 10 min.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	To assay cell invasion, 500 μL culture supernatant was collected from transfected cells and added to the upper Transwell chamber.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	GIST cells (2 × 10 cells) in about 200 μL serum-free medium were added to the lower chamber.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The cells were cultured for 2 d at 37 °C with 5% CO2.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	After culturing, cells remaining in the lower chamber were removed with cotton swabs and those in the upper chamber were stained with 0.2% crystal violet for 5 min.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	We used an inverted microscope to count the cells that had migrated through the membrane and invaded the upper chamber.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	We bought 5-wk-old; male BALB/c nude mice from Vital River Laboratories (Beijing, China) and housed them for 1 wk to adapt to the environment.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	GIST-T1 cells (5 × 10) transfected with OE-IGF2/OE-NC, sh-IGF2/sh-NC were injected into the inguinal skin and the mice were monitored for growth of the tumor for 7 d before being randomized to four groups and treated with imatinib 50 mg/kg daily.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	After 4 wk, we killed the mice with an overdose of pentobarbital.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	All animal experiments were approved by the Animal Ethics Committee of Beijing Viewsolid Biotechnology Co. LTD (Protocol No. VS2126A00170) and all methods followed the ARRIVE guidelines.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	We fixed the liver tissue of mice in neutral formalin (10%), embedded it in paraffin, cut the tissue into 4 µm sections, and stained it with hematoxylin and eosin (HE).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The sections were observed with a microscope.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Cells were incubated in commercial seahorse XF assay medium plus pyruvate (1 mmol/L), glucose (10 mmol/L) and glutamine (2 mmol/L) 37 °C for 1 h in a CO2-free incubator.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The rate of extracellular acidification was measured before and after addition of oligomycin, glucose, and 2-deoxy-D-glucose (2-DG).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	FCCP, a mitochondrial uncoupling agent; oligomycin, an ATP synthase inhibitor; 2-DG, a glycolysis inhibitor; rotenone; and antimycin A were added and metabolic energy consumption was assayed with a Seahorse XF96 Analyzer (Agilent, Santa Clara, CA, United States).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The concentration of lactate in transfected cells was determined by ELISA with lactate assay kits (MAK064; Sigma-Aldrich, St Louis, MO, United States) according to the manufacturer’s protocol.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The optical density of each well was determined at 570 nm (Plate Reader AF2000; Eppendorf, Waltham, MA, United States).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	GIST cell apoptosis was assayed by flow cytometry (LSRII; BD Biosciences, Franklin Lakes, NJ, United States).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	using annexin V-FITC apoptosis detection kits.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The apoptosis rate was determined by analysis of Q2 and Q3 quadrant cells.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	We used GraphPad Prism 7.0 for data analysis.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Data were reported as mean ± standard deviation of three independent experiments.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Single-group comparisons were done with Student’s t-tests.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Multiple group differences were compared by analysis of variance.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	P < 0.05 indicated significance.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Based on the limma R package, a total of 2578 (DEGs 1398 downregulated and 1188 upregulated) were screened out from GEO: GSE225819 data, including 20 normal samples and 20 GIST samples with liver metastasis (\|log2FC\| > 1; P < 0.05), suggesting that these DEGs may be involved in liver metastasis in GIST patients (Figure 1A).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The top 10 upregulated genes were PENK, IGF2, GPR20, CTSL, SCRG1, PNMAL1, NKX3-2, ANO1, PLAT, and BCHE.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The top 10 downregulated genes were ATP4B, GKN1, MT1G, GKN2, ATP4A, SPINK1, TSPAN8, TFF1, KCNE2, and REG1A (Supplementary Table 1).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Based on the Deseq2, 1386 DEGs (939 downregulated and 447 upregulated) were screened out in GSE155880, including seven Imatinib-sensitive samples and seven imatinib-resistant GIST patients (\|log2FC\| > 1; P < 0.05, Figure 1B).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The intersection of the two analyses indicated that only IGF2 was involved in the drug resistance regulation and GIST metastasis in these DEGs (Supplementary Table 2).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Moreover, we evaluated IGF2 expression in the GIST cell line.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	By western blotting, expression levels of IGF2 in GIST882, GIST882-R, GIST-T1, and GIST-T1-R were higher than those in normal RGM-1.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Furthermore, IGF2 was significantly over expressed in GIST882-R/GIST-T1-R compared with other cell lines GIST882/GIST-T1 (P < 0.01, P < 0.001; Figure 1C).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	In addition, the expression levels of IGF2 in culture supernatants were measured using ELISA and compared (Figure 1D).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	We found that the ELISA and western blot results (P < 0.05, P < 0.001) were similar.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	IGF2 expression was high in drug-resistant GIST cell lines, suggesting that IGF2 overexpression may be closely related to drug resistance.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	High expression of insulin-like growth factor 2 in gastrointestinal stromal tumors with liver metastasis and closely related to drug resistance.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	A: Differentially expressed genes in gastrointestinal stromal tumors (GIST) with liver metastasis tissues and normal gastric tissues (\|log2FC\| > 1; P < 0.05); B: Differentially expressed genes in imatinib sensitive and in seven Imatinib-resistant GIST patients (\|log2FC\| > 1; P < 0.05); C: Western blot assay of insulin-like growth factor 2 (IGF2) protein expression in GIST cell lines (GIST882, GIST882-R, GIST-T1, GIST-T1-R); D: ELISA of IGF2 expression in GIST cell lines (GIST882, GIST882-R, GIST-T1, GIST-T1-R).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Data are mean ± standard deviation.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	P < 0.05; P < 0.01; P < 0.001.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	We transfected GIST882 and GIST-T1 cells with an IGF2 overexpressing plasmid (OE-IGF2) or a shRNA to knock down IGF2 (sh-IGF2).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Western blotting detected the efficiency of cell transfection (Figure 2A).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	IGF2 was highly expressed in OE-IGF2-transfected cells compared with OE-NC cells, while IGF2 expression was low in sh-IGF2-transfected cells (P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	ELISA also found that IGF2 expression high in OE-IGF2 group compared with OE-NC-GIST882 and GIST-T1 cells and IGF2 was low expressed in sh-IGF2-transfected cells (Figure 2B, P < 0.05, P < 0.01, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The CCK-8 results showed that cell viability was significantly increased after exogenous expression of IGF2, sh-IGF2 transfection inhibited GIST882 and GIST-T1 cell viability (Figure 2C, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Likewise, the Transwell assays found more migrating and invading OE-IGF2-GIST882 and GIST-T1 cells compared with their respective control cells (Figure 2D and E, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	We also found that sh-IGF2 transfection inhibited cell viability, migration and invasion.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	In addition, western blotting detect EMT-related proteins (E-cadherin, vimentin, Twist1, and N-cadherin) expression in cells.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Silencing IGF2 increased E-cadherin expression, and inhibited vimentin, Twist1, and N-cadherin expression, but IGF2 overexpression had the opposite experimental findings (Figure 2F, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	To further verify the functional role of IGF2 on the growth of GISTs, we performed nude mouse tumorigenesis experiments.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	OE-IGF2 transfected-GIST-T1 cell lines were injected into the spleen.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	We found that OE-IGF2 promoted the GIST-T1 cell metastasis in vivo, showing a significant decline in the number of liver metastatic nodules (Figure 2G and H, P < 0.01).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Insulin-like growth factor 2 promotes malignant characteristics and metastasis of gastrointestinal stromal tumors.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	A: Western blot measured the transfection efficiency of OE-insulin-like growth factor 2 (IGF2) or sh-IGF2 in gastrointestinal stromal tumors (GIST) 882 and GIST-T1 cells; B: ELISA of IGF2 expression in OE-IGF2 or sh-IGF2 transfected GIST882 and GIST-T1 cells; C: Cell counting kit-8 assay assessed cell viability in GIST882 and GIST-T1 cells; D: Transwell assay evaluated the migration of OE-IGF2- or sh-IGF2-transfected cells (scar bar = 50 μm); E: Transwell assays of the invasiveness of OE-IGF2 or sh-IGF2 transfected cells. (
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	scar bar = 50 μm); F: Detection of proteins involved in epithelial-mesenchymal transition (vimentin, N-cadherin, E-cadherin, Twist1) in OE-IGF2 or sh-IGF2 transfected cells; G: Liver tissue from tumor xenografts in nude mice injected withOE-IGF2 transfected GIST-T1 cells; H: Liver metastasis determined by hematoxylin-eosin staining.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Data are mean ± standard deviation.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	P < 0.05; P < 0.01; P < 0.001.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	IGF1R mRNA expression was increased in GIST-T1 and GIST882 cells transfected with OE-IGF2, and IGF1R mRNA expression was decreased after sh-IGF2 transfection (Figure 3A, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	PI3K-Akt signaling is the IGF2-IGF1R signal principal downstream target.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Expression of IGF2-IGF1R pathway-associated proteins (IGF1R, p-IGF1R, PI3K, AKT, p-AKT) in GIST-T1 cells was measured by western blotting.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	IGF2 overexpression increased the expression of IGF1R, p-IGF1R, PI3K, AKT, and p-AKT in GIST-T1 cells.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The opposite result was noted after IGF2 knockdown (Figure 3B, P < 0.01, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Although sh-IGF2 reduced IGF1R, p-IGF1R, PI3K, AKT, and p-AKT expression in GIST-T1 cells, it was partially restored by overexpression of IGF2R (Figure 3C, P < 0.01, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Insulin-like growth factor 2 activated the IGF1R signaling in gastrointestinal stromal tumors cells.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	A: Quantitative reverse transcriptase PCR assay of IGF1R mRNA expression in gastrointestinal stromal tumors (GIST) 882 and GIST-T1 cells after OE-insulin-like growth factor 2 (IGF2) or sh-IGF2 transfection; B: Detection of protein levels (IGF1R, p-IGF1R, PI3K, AKT, and p-AKT) involved in the PI3K/AKT in OE-IGF2 or sh-IGF2 transfected-GIST-T1 cells by western blot assay; C: Detection of protein levels (IGF1R, p-IGF1R, PI3K, AKT, and p-AKT) involved in the PI3K/AKT in GIST-T1 cells after sh-IGF2 and OE-IGF2R transfection by western blot assay.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	P < 0.01; P < 0.001.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	We analyzed glucose consumption and lactate production in GIST cells.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Sh-IGF2 inhibited glucose consumption (Figure 4A), and lactate production in GIST882 and GIST-T1 cells (Figure 4B), but IGF2 overexpression had the opposite experimental findings (P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	To examine the role of the Warburg effect in liver metastasis of GISTs, we treated OE-NC-GIST882 and OE-IGF2-GIST882 cells with 2-deoxyglucose (2-DG, a glycolysis inhibitor) for 24 hat 0, 4, 8, and 16 mmol/L. 2-DG significantly inhibited glycolysis (Figure 4C, P < 0.05, P < 0.01, P < 0.001) and Transwell assays found that 2-DG treatment inhibited the promoting effect of OE-IGF2 on GIST882 and GIST-T1 cell invasion and migration (Figure 4D and E, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Similarly, OE-IGF2 increased vimentin, Twist1, and N-cadherin expression and inhibited E-cadherin expression in cells, but the expression was partially restored by 2-DG treatment (Figure 4F, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Insulin-like growth factor 2/IGF1R-mediatedglycolysisis required for gastrointestinal stromal tumors with liver metastasis.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	A: Extracellular acidification rate was measured; B: Lactate production in gastrointestinal stromal tumors (GIST) 882 and GIST-T1 cells transfected with sh-insulin-like growth factor 2 (IGF2) or OE-IGF2 were measured; C: Lactate production in OE-IGF2-GIST882 and GIST-T1 cells cotreated with 2-deoxy-D-glucose (2-DG) (0, 4, 8, and 16 mmol/L); D: Transwell assay of the migration ability of the OE-IGF2-cells cotreated with 2-DG (scar bar = 50 μm); E: Transwell assay of the invasiveness of OE-IGF2-cells cotreated with 2-DG (scar bar = 50 μm); F: Assay of proteins involved in epithelial-mesenchymal transition (vimentin, N-cadherin, E-cadherin, Twist1) in OE-IGF2-cells cotreated with 2-DG.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Data are mean ± standard deviation.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	P < 0.05; P < 0.01; P < 0.001.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Figure 1 shows that IGF2 was involved in regulating drug resistance.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Next, we will further verify.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	To test whether IGF2 also regulated drug resistance in GISTs in vivo, we established a xenograft model by inoculating sh-NC or sh-IGF2-GIST-T1 cells into nude mice.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	In the sh-IGF2-GIST-T1 mouse xenograft model, tumor volume and growth were inhibited by sh-IGF2, and imatinib had the same influence on tumor growth and volume.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Combined treatment with imatinib and sh-IGF2 was more effective for reducing tumor progression than single treatment (Figure 5A-C, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The western blot results revealed that expression of IGF1R, p-IGF1R, AKT, PI3K, and p-AKT in tumor tissue was suppressed in both sh-IGF2-transfected cells and after imatinib treatment.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Moreover, combined imatinib and sh-IGF2 were more effective than single therapy (Figure 5D, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	The above data suggest that IGF2/IGF1R regulate imatinib resistance.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Insulin-like growth factor 2/IGF1R regulates imatinib resistance of gastrointestinal stromal tumors by regulating glycolysis.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	A: Tumor growth in xenografted nude mice; B: Tumor volumes in sh-insulin-like growth factor 2 (IGF2)-gastrointestinal stromal tumors (GIST)-T1 mouse xenograft models treated with imatinib; C: After 35 d, the mice were killed and the tumors were weighed; D: Assay of IGF1R, p-IGF1R, PI3K, AKT, and p-AKT in tumor tissue by western blotting; E: Assay of drug sensitivity in OE-IGF2-GIST882 and GIST-T1 cells treated with 2-deoxy-D-glucose (2-DG); F: Flow cytometry assay of apoptosis of OE-IGF2-GIST882 and GIST-T1 cells treated with 2-DG.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Data are mean ± standard deviation.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	P < 0.05; P < 0.01; P < 0.001.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	In addition, previous data shows that IGF2 regulates glycolysis in GIST cells.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	IGF2 regulates cell sensitivity to imatinib through its influence on glycolysis.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	We used 2-DG to inhibit glycolysis in GIST cells.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	OE-IGF2 increased drug sensitivity in GIST882 and GIST-T1 cells, but after treatment with 2-DG, transfection with OE-IGF2 no longer changed drug sensitivity in GIST cells (Figure 5E, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Flow cytometric analysis showed that sh-IGF2 suppressed imatinib-induced apoptosis and OE-IGF2 reduced imatinib-induced apoptosis in GIST cells.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Treatment with 2-DG and transfection with OE-IGF2 no longer influenced imatinib-induced apoptosis in GIST cells (Figure 5F, P < 0.001).
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Therefore, the results show that IGF2 regulated imatinib sensitivity in GIST cells by affecting glycolysis.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	GISTs is the most frequent malignant gastrointestinal sarcoma and causes significant patient harm.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Recently, anticancer drug resistance has become a significant challenge to the treatment of GISTs.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	Treatment with tyrosine kinase inhibitors (TKIs) has led to substantial improvement of survival, both for patients with localized GISTs and those with advanced disease.
PMC11334037	Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors	As the first-line TKI, imatinib offers treatment for advanced and metastatic GISTs, adjuvant therapy in high-risk GISTs and neoadjuvant treatment to downsize large tumors prior to resection.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

After overnight incubation, the cells were treated with imatinib at 0, 20, 40, 60, and 80 μmol/L for 48 h. CompuSyn software was used to calculate the combination index using the Chou-Talalay method to determine the antagonistic influence.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

For the migration assay, GIST cells were seeded into 8 µm well Transwell chambers (Corning; Corning, NY, United States).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The upper chamber was filled with 200 µL serum-free medium containing 2 × 10 cells and the lower chamber was filled with 500 μL complete medium (10% FBS).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

After 48 h, the cells were fixed with formaldehyde and stained with 0.2% crystal violet for 10 min.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

To assay cell invasion, 500 μL culture supernatant was collected from transfected cells and added to the upper Transwell chamber.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

GIST cells (2 × 10 cells) in about 200 μL serum-free medium were added to the lower chamber.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The cells were cultured for 2 d at 37 °C with 5% CO2.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

After culturing, cells remaining in the lower chamber were removed with cotton swabs and those in the upper chamber were stained with 0.2% crystal violet for 5 min.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

We used an inverted microscope to count the cells that had migrated through the membrane and invaded the upper chamber.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

We bought 5-wk-old; male BALB/c nude mice from Vital River Laboratories (Beijing, China) and housed them for 1 wk to adapt to the environment.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

GIST-T1 cells (5 × 10) transfected with OE-IGF2/OE-NC, sh-IGF2/sh-NC were injected into the inguinal skin and the mice were monitored for growth of the tumor for 7 d before being randomized to four groups and treated with imatinib 50 mg/kg daily.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

After 4 wk, we killed the mice with an overdose of pentobarbital.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

All animal experiments were approved by the Animal Ethics Committee of Beijing Viewsolid Biotechnology Co. LTD (Protocol No. VS2126A00170) and all methods followed the ARRIVE guidelines.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

We fixed the liver tissue of mice in neutral formalin (10%), embedded it in paraffin, cut the tissue into 4 µm sections, and stained it with hematoxylin and eosin (HE).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The sections were observed with a microscope.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Cells were incubated in commercial seahorse XF assay medium plus pyruvate (1 mmol/L), glucose (10 mmol/L) and glutamine (2 mmol/L) 37 °C for 1 h in a CO2-free incubator.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The rate of extracellular acidification was measured before and after addition of oligomycin, glucose, and 2-deoxy-D-glucose (2-DG).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

FCCP, a mitochondrial uncoupling agent; oligomycin, an ATP synthase inhibitor; 2-DG, a glycolysis inhibitor; rotenone; and antimycin A were added and metabolic energy consumption was assayed with a Seahorse XF96 Analyzer (Agilent, Santa Clara, CA, United States).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The concentration of lactate in transfected cells was determined by ELISA with lactate assay kits (MAK064; Sigma-Aldrich, St Louis, MO, United States) according to the manufacturer’s protocol.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The optical density of each well was determined at 570 nm (Plate Reader AF2000; Eppendorf, Waltham, MA, United States).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

GIST cell apoptosis was assayed by flow cytometry (LSRII; BD Biosciences, Franklin Lakes, NJ, United States).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

using annexin V-FITC apoptosis detection kits.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The apoptosis rate was determined by analysis of Q2 and Q3 quadrant cells.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

We used GraphPad Prism 7.0 for data analysis.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Data were reported as mean ± standard deviation of three independent experiments.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Single-group comparisons were done with Student’s t-tests.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Multiple group differences were compared by analysis of variance.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

P < 0.05 indicated significance.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Based on the limma R package, a total of 2578 (DEGs 1398 downregulated and 1188 upregulated) were screened out from GEO: GSE225819 data, including 20 normal samples and 20 GIST samples with liver metastasis (|log2FC| > 1; P < 0.05), suggesting that these DEGs may be involved in liver metastasis in GIST patients (Figure 1A).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The top 10 upregulated genes were PENK, IGF2, GPR20, CTSL, SCRG1, PNMAL1, NKX3-2, ANO1, PLAT, and BCHE.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The top 10 downregulated genes were ATP4B, GKN1, MT1G, GKN2, ATP4A, SPINK1, TSPAN8, TFF1, KCNE2, and REG1A (Supplementary Table 1).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Based on the Deseq2, 1386 DEGs (939 downregulated and 447 upregulated) were screened out in GSE155880, including seven Imatinib-sensitive samples and seven imatinib-resistant GIST patients (|log2FC| > 1; P < 0.05, Figure 1B).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The intersection of the two analyses indicated that only IGF2 was involved in the drug resistance regulation and GIST metastasis in these DEGs (Supplementary Table 2).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Moreover, we evaluated IGF2 expression in the GIST cell line.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

By western blotting, expression levels of IGF2 in GIST882, GIST882-R, GIST-T1, and GIST-T1-R were higher than those in normal RGM-1.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Furthermore, IGF2 was significantly over expressed in GIST882-R/GIST-T1-R compared with other cell lines GIST882/GIST-T1 (P < 0.01, P < 0.001; Figure 1C).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

In addition, the expression levels of IGF2 in culture supernatants were measured using ELISA and compared (Figure 1D).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

We found that the ELISA and western blot results (P < 0.05, P < 0.001) were similar.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

IGF2 expression was high in drug-resistant GIST cell lines, suggesting that IGF2 overexpression may be closely related to drug resistance.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

High expression of insulin-like growth factor 2 in gastrointestinal stromal tumors with liver metastasis and closely related to drug resistance.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

A: Differentially expressed genes in gastrointestinal stromal tumors (GIST) with liver metastasis tissues and normal gastric tissues (|log2FC| > 1; P < 0.05); B: Differentially expressed genes in imatinib sensitive and in seven Imatinib-resistant GIST patients (|log2FC| > 1; P < 0.05); C: Western blot assay of insulin-like growth factor 2 (IGF2) protein expression in GIST cell lines (GIST882, GIST882-R, GIST-T1, GIST-T1-R); D: ELISA of IGF2 expression in GIST cell lines (GIST882, GIST882-R, GIST-T1, GIST-T1-R).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Data are mean ± standard deviation.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

P < 0.05; P < 0.01; P < 0.001.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

We transfected GIST882 and GIST-T1 cells with an IGF2 overexpressing plasmid (OE-IGF2) or a shRNA to knock down IGF2 (sh-IGF2).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Western blotting detected the efficiency of cell transfection (Figure 2A).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

IGF2 was highly expressed in OE-IGF2-transfected cells compared with OE-NC cells, while IGF2 expression was low in sh-IGF2-transfected cells (P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

ELISA also found that IGF2 expression high in OE-IGF2 group compared with OE-NC-GIST882 and GIST-T1 cells and IGF2 was low expressed in sh-IGF2-transfected cells (Figure 2B, P < 0.05, P < 0.01, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The CCK-8 results showed that cell viability was significantly increased after exogenous expression of IGF2, sh-IGF2 transfection inhibited GIST882 and GIST-T1 cell viability (Figure 2C, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Likewise, the Transwell assays found more migrating and invading OE-IGF2-GIST882 and GIST-T1 cells compared with their respective control cells (Figure 2D and E, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

We also found that sh-IGF2 transfection inhibited cell viability, migration and invasion.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

In addition, western blotting detect EMT-related proteins (E-cadherin, vimentin, Twist1, and N-cadherin) expression in cells.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Silencing IGF2 increased E-cadherin expression, and inhibited vimentin, Twist1, and N-cadherin expression, but IGF2 overexpression had the opposite experimental findings (Figure 2F, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

To further verify the functional role of IGF2 on the growth of GISTs, we performed nude mouse tumorigenesis experiments.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

OE-IGF2 transfected-GIST-T1 cell lines were injected into the spleen.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

We found that OE-IGF2 promoted the GIST-T1 cell metastasis in vivo, showing a significant decline in the number of liver metastatic nodules (Figure 2G and H, P < 0.01).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Insulin-like growth factor 2 promotes malignant characteristics and metastasis of gastrointestinal stromal tumors.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

A: Western blot measured the transfection efficiency of OE-insulin-like growth factor 2 (IGF2) or sh-IGF2 in gastrointestinal stromal tumors (GIST) 882 and GIST-T1 cells; B: ELISA of IGF2 expression in OE-IGF2 or sh-IGF2 transfected GIST882 and GIST-T1 cells; C: Cell counting kit-8 assay assessed cell viability in GIST882 and GIST-T1 cells; D: Transwell assay evaluated the migration of OE-IGF2- or sh-IGF2-transfected cells (scar bar = 50 μm); E: Transwell assays of the invasiveness of OE-IGF2 or sh-IGF2 transfected cells. (

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

scar bar = 50 μm); F: Detection of proteins involved in epithelial-mesenchymal transition (vimentin, N-cadherin, E-cadherin, Twist1) in OE-IGF2 or sh-IGF2 transfected cells; G: Liver tissue from tumor xenografts in nude mice injected withOE-IGF2 transfected GIST-T1 cells; H: Liver metastasis determined by hematoxylin-eosin staining.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Data are mean ± standard deviation.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

P < 0.05; P < 0.01; P < 0.001.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

IGF1R mRNA expression was increased in GIST-T1 and GIST882 cells transfected with OE-IGF2, and IGF1R mRNA expression was decreased after sh-IGF2 transfection (Figure 3A, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

PI3K-Akt signaling is the IGF2-IGF1R signal principal downstream target.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Expression of IGF2-IGF1R pathway-associated proteins (IGF1R, p-IGF1R, PI3K, AKT, p-AKT) in GIST-T1 cells was measured by western blotting.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

IGF2 overexpression increased the expression of IGF1R, p-IGF1R, PI3K, AKT, and p-AKT in GIST-T1 cells.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The opposite result was noted after IGF2 knockdown (Figure 3B, P < 0.01, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Although sh-IGF2 reduced IGF1R, p-IGF1R, PI3K, AKT, and p-AKT expression in GIST-T1 cells, it was partially restored by overexpression of IGF2R (Figure 3C, P < 0.01, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Insulin-like growth factor 2 activated the IGF1R signaling in gastrointestinal stromal tumors cells.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

A: Quantitative reverse transcriptase PCR assay of IGF1R mRNA expression in gastrointestinal stromal tumors (GIST) 882 and GIST-T1 cells after OE-insulin-like growth factor 2 (IGF2) or sh-IGF2 transfection; B: Detection of protein levels (IGF1R, p-IGF1R, PI3K, AKT, and p-AKT) involved in the PI3K/AKT in OE-IGF2 or sh-IGF2 transfected-GIST-T1 cells by western blot assay; C: Detection of protein levels (IGF1R, p-IGF1R, PI3K, AKT, and p-AKT) involved in the PI3K/AKT in GIST-T1 cells after sh-IGF2 and OE-IGF2R transfection by western blot assay.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

P < 0.01; P < 0.001.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

We analyzed glucose consumption and lactate production in GIST cells.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Sh-IGF2 inhibited glucose consumption (Figure 4A), and lactate production in GIST882 and GIST-T1 cells (Figure 4B), but IGF2 overexpression had the opposite experimental findings (P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

To examine the role of the Warburg effect in liver metastasis of GISTs, we treated OE-NC-GIST882 and OE-IGF2-GIST882 cells with 2-deoxyglucose (2-DG, a glycolysis inhibitor) for 24 hat 0, 4, 8, and 16 mmol/L. 2-DG significantly inhibited glycolysis (Figure 4C, P < 0.05, P < 0.01, P < 0.001) and Transwell assays found that 2-DG treatment inhibited the promoting effect of OE-IGF2 on GIST882 and GIST-T1 cell invasion and migration (Figure 4D and E, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Similarly, OE-IGF2 increased vimentin, Twist1, and N-cadherin expression and inhibited E-cadherin expression in cells, but the expression was partially restored by 2-DG treatment (Figure 4F, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Insulin-like growth factor 2/IGF1R-mediatedglycolysisis required for gastrointestinal stromal tumors with liver metastasis.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

A: Extracellular acidification rate was measured; B: Lactate production in gastrointestinal stromal tumors (GIST) 882 and GIST-T1 cells transfected with sh-insulin-like growth factor 2 (IGF2) or OE-IGF2 were measured; C: Lactate production in OE-IGF2-GIST882 and GIST-T1 cells cotreated with 2-deoxy-D-glucose (2-DG) (0, 4, 8, and 16 mmol/L); D: Transwell assay of the migration ability of the OE-IGF2-cells cotreated with 2-DG (scar bar = 50 μm); E: Transwell assay of the invasiveness of OE-IGF2-cells cotreated with 2-DG (scar bar = 50 μm); F: Assay of proteins involved in epithelial-mesenchymal transition (vimentin, N-cadherin, E-cadherin, Twist1) in OE-IGF2-cells cotreated with 2-DG.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Data are mean ± standard deviation.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

P < 0.05; P < 0.01; P < 0.001.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Figure 1 shows that IGF2 was involved in regulating drug resistance.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Next, we will further verify.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

To test whether IGF2 also regulated drug resistance in GISTs in vivo, we established a xenograft model by inoculating sh-NC or sh-IGF2-GIST-T1 cells into nude mice.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

In the sh-IGF2-GIST-T1 mouse xenograft model, tumor volume and growth were inhibited by sh-IGF2, and imatinib had the same influence on tumor growth and volume.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Combined treatment with imatinib and sh-IGF2 was more effective for reducing tumor progression than single treatment (Figure 5A-C, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The western blot results revealed that expression of IGF1R, p-IGF1R, AKT, PI3K, and p-AKT in tumor tissue was suppressed in both sh-IGF2-transfected cells and after imatinib treatment.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Moreover, combined imatinib and sh-IGF2 were more effective than single therapy (Figure 5D, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

The above data suggest that IGF2/IGF1R regulate imatinib resistance.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Insulin-like growth factor 2/IGF1R regulates imatinib resistance of gastrointestinal stromal tumors by regulating glycolysis.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

A: Tumor growth in xenografted nude mice; B: Tumor volumes in sh-insulin-like growth factor 2 (IGF2)-gastrointestinal stromal tumors (GIST)-T1 mouse xenograft models treated with imatinib; C: After 35 d, the mice were killed and the tumors were weighed; D: Assay of IGF1R, p-IGF1R, PI3K, AKT, and p-AKT in tumor tissue by western blotting; E: Assay of drug sensitivity in OE-IGF2-GIST882 and GIST-T1 cells treated with 2-deoxy-D-glucose (2-DG); F: Flow cytometry assay of apoptosis of OE-IGF2-GIST882 and GIST-T1 cells treated with 2-DG.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Data are mean ± standard deviation.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

P < 0.05; P < 0.01; P < 0.001.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

In addition, previous data shows that IGF2 regulates glycolysis in GIST cells.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

IGF2 regulates cell sensitivity to imatinib through its influence on glycolysis.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

We used 2-DG to inhibit glycolysis in GIST cells.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

OE-IGF2 increased drug sensitivity in GIST882 and GIST-T1 cells, but after treatment with 2-DG, transfection with OE-IGF2 no longer changed drug sensitivity in GIST cells (Figure 5E, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Flow cytometric analysis showed that sh-IGF2 suppressed imatinib-induced apoptosis and OE-IGF2 reduced imatinib-induced apoptosis in GIST cells.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Treatment with 2-DG and transfection with OE-IGF2 no longer influenced imatinib-induced apoptosis in GIST cells (Figure 5F, P < 0.001).

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Therefore, the results show that IGF2 regulated imatinib sensitivity in GIST cells by affecting glycolysis.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

GISTs is the most frequent malignant gastrointestinal sarcoma and causes significant patient harm.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Recently, anticancer drug resistance has become a significant challenge to the treatment of GISTs.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

Treatment with tyrosine kinase inhibitors (TKIs) has led to substantial improvement of survival, both for patients with localized GISTs and those with advanced disease.

PMC11334037

Insulin-like growth factor 2 targets IGF1R signaling transduction to facilitate metastasis and imatinib resistance in gastrointestinal stromal tumors

As the first-line TKI, imatinib offers treatment for advanced and metastatic GISTs, adjuvant therapy in high-risk GISTs and neoadjuvant treatment to downsize large tumors prior to resection.