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PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Twenty-four 4-weeks-old BALB/c nude mice (Vital River Laboratory Animal Co., Ltd., Beijing, China) were animal experiment subjects.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
The experiments were approved by the Institutional Animal Care and Use Committee of the Affiliated Hospital of Inner Mongolia Minzu University (n° 201306YT2).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
All animal experiments complied with the ARRIVE guidelines.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
GIST-882 cells (1×10) stably transfected with si-NC and si-circAPP were subcutaneously inoculated into BALB/c nude mice (n = 6/group), and tumor volumes were measured weekly with vernier calipers from the second week after injection. .
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Five weeks later, all mice were euthanized by cervical dislocation under isoflurane (5 %) treatment.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Tumors were resected for immunohistochemistry with antibodies against HK2 (22029-1-AP, Proteintech), PKM2 (15822-1-AP, Proteintech), and Ki-67 (ab15580, Abcam).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
To determine the distant metastasis of tumors, GIST-882 cells (1×10) stably transfected with si-NC and si-circAPP were injected into nude mice via tail vein, and the nude mice were euthanized after 8 weeks to assess liver metastasis by HE-staining.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
All statistical analyses were performed using GraphPad Prism 9.0 (GraphPad Software, CA, USA).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Data represent mean ± Standard Deviation (SD).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Two-tailed Student's t-test and one-way ANOVA determined statistical differences.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Statistical significance represented p < 0.05.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
CircAPP has been revealed to be abnormally expressed in GIST.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Consistent results were also obtained in this study, and circAPP expression in GIST tissues and cell lines was higher than that in normal tissues and cells (Fig. 1A, B).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
The circbase database discovered that circAPP is 678 bp in length and consisted of exons 4‒7 of the APP gene located on chromosome 21q21.3 (Fig. 1C).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Examinations of the ring structure of circAPP revealed that RNase R treatment only decreased linear APP mRNA but not circAPP (Fig. 1D).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
After actinomycin D treatment, circAPP had a longer half-life than linear APP mRNA (Fig. 1E).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Subsequently, the location of circAPP in GIST-882 cells was examined by subcellular isolation assay, confirming circAPP mainly expressed in the cytoplasm (Fig. 1F).Fig.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
1CircAPP is up-regulated in GIST. (
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
CircAPP in GIST tissues and adjacent normal tissues, (B) CircAPP in GIST cell lines and normal cell lines, (C) Gene structure of circAPP on the circbase database, (D and E) RNaseR and actinomycin D treatment to explore the ring structure of circAPP, (F) Subcellular localization of circAPP; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.Fig 1 CircAPP is up-regulated in GIST. (
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
CircAPP in GIST tissues and adjacent normal tissues, (B) CircAPP in GIST cell lines and normal cell lines, (C) Gene structure of circAPP on the circbase database, (D and E) RNaseR and actinomycin D treatment to explore the ring structure of circAPP, (F) Subcellular localization of circAPP; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Since GIST-882 has the highest expression level of circAPP, follow-up studies were performed using GIST-882.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Focusing on the functions of circAPP in GIST, si-circAPP was transected into GIST-882 cells which maintained circAPP expression in a suppressed state (Fig. 2A).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Corresponding to the decrease in circAPP expression, the cloning ability and Edu-positive cell number of GIST-882 cells examined by colony formation assay (Fig. 2B) and EdU assay (Fig. 2C) were reduced, and the apoptosis rate tested by flow cytometry was augmented (Fig. 2D).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
For examining Warburg effect in GIST-882 cells, glucose consumption, lactate production, and ATP levels, along with ATP/ADP and NAD/NADH were the tested indices.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
In circAPP-silenced GIST-882 cells, reduction in glucose consumption, lactate production, ATP, and ATP/ADP, and upregulation of NAD/NADH were detectable (Fig. 2E, F).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Moreover, key Warburg effect proteins HK2 and PKM2 were examined by western blot: after silencing circAPP, HK2 and PKM2 levels were reduced (Fig. 2G), further proving the anti-Warburg effect property of silencing circAPP in GIST.Fig.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
2Knockdown of circAPP inhibits GIST proliferation and the Warburg effect.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
si-circAPP was transfected into GIST-882 and GES-1 cells. (
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Expression changes of circAPP, (B and C) Colony formation and EdU assays to assess cell proliferation, (D) Flow cytometry to analyze apoptosis, (E) Cellular glucose consumption, lactate production, and ATP levels, (F) ATP/ADP ratio and NAD/NADH ratio, (G) Western blot to evaluate cellular HK2 and PKM2 protein expression; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.Fig 2 Knockdown of circAPP inhibits GIST proliferation and the Warburg effect.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
si-circAPP was transfected into GIST-882 and GES-1 cells. (
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Expression changes of circAPP, (B and C) Colony formation and EdU assays to assess cell proliferation, (D) Flow cytometry to analyze apoptosis, (E) Cellular glucose consumption, lactate production, and ATP levels, (F) ATP/ADP ratio and NAD/NADH ratio, (G) Western blot to evaluate cellular HK2 and PKM2 protein expression; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Through the prediction on the bioinformatics website https://starbase.sysu.edu.cn/, circAPP had a potential binding site for miR-6838-5p (Fig. 3A).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Subsequently, this targeting relationship was confirmed, as the results claimed that luciferase activity decreased after co-transfection of WT-circAPP and miR-6838-5p mimic (Fig. 3B); miR-6838-5p and circAPP increased after Ago2 treatment (Fig. 3C).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
RNA gene analysis manifested the reduction of miR-6838-5p in GIST tissues and cells (Fig. 3D, E).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Furthermore, the knockdown of circAPP in GIST-882 cells was also found to promote miR-6838-5p expression (Fig. 3F).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Taken together, circAPP targets the regulation of miR-6838-5p levels.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Fig. 3CircAPP has a binding relationship with miR-6838-5p. (
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Bioinformatics website to predict the potential targets of circAPP and miR-6838-5p, (B and C) Dual luciferase reporter assay and RIP assay to detect the targeting relationship between circAPP and miR-6838-5p, (D and E) miR-6838-5p expression in tissues and cells, (F) Effect of knockdown of circAPP on miR-6838-5p expression; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.Fig 3 CircAPP has a binding relationship with miR-6838-5p. (
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Bioinformatics website to predict the potential targets of circAPP and miR-6838-5p, (B and C) Dual luciferase reporter assay and RIP assay to detect the targeting relationship between circAPP and miR-6838-5p, (D and E) miR-6838-5p expression in tissues and cells, (F) Effect of knockdown of circAPP on miR-6838-5p expression; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
To confirm circAPP/miR-838-5p regulating the biological behavior of GIST, miR-6838-5p inhibitor and si-circAPP were transfected into GIST-882 cells.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Transfection of si-circAPP alone miR-6838-5p expression, but the action of si-circAPP was impaired by miR-6838-5p inhibitor (Fig. 4A).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Functional experiments showed (Fig. 4B‒G) that transfection of miR-6838-5p inhibitor alone enhanced the clonality and proliferation, decreased apoptosis rate, and promoted cellular glucose consumption, lactate production, ATP and ATP/ADP, downregulated NAD/NADH, and induced HK2 and PKM2 protein expression.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
si-circAPP showed opposite effects which could be blocked by co-transfection of miR-6838-5p inhibitor.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
All in all, circAPP affects GIST proliferation and the Warburg effect by regulating miR-6838-5p expression.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Fig. 4miR-6838-5p inhibitor promotes GIST proliferation and the Warburg effect and reverses the effect of si-circAPP.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
miR-6838-5p inhibitor was transfected alone or si-circAPP and miR-6838-5p inhibitor were co-transfected, (A) Expression changes of miR-6838-5p, (B and C) Colony formation and EdU assays to assess cell proliferation, (D) Flow cytometry to analyze apoptosis, (E) Cellular glucose consumption, lactate production, and ATP levels, (F) ATP/ADP ratio and NAD/NADH ratio, (G) Western blot to evaluate cellular HK2 and PKM2 protein expression; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.Fig 4 miR-6838-5p inhibitor promotes GIST proliferation and the Warburg effect and reverses the effect of si-circAPP.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
miR-6838-5p inhibitor was transfected alone or si-circAPP and miR-6838-5p inhibitor were co-transfected, (A) Expression changes of miR-6838-5p, (B and C) Colony formation and EdU assays to assess cell proliferation, (D) Flow cytometry to analyze apoptosis, (E) Cellular glucose consumption, lactate production, and ATP levels, (F) ATP/ADP ratio and NAD/NADH ratio, (G) Western blot to evaluate cellular HK2 and PKM2 protein expression; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
The bioinformatics website starbase predicted the existence of potential binding sites for miR-6838-5p and CDV3 (Fig. 5A).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Co-transfection of WT-CDV3 and miR-6838-5p mimic inhibited luciferase activity (Fig. 5B); CDV3 and miR-6838-5p were enriched with Ago2 (Fig. 5C).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
CDV3 was higher in both GIST tissues and cell lines than in normal tissues and cell lines (Fig. 5D, E).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Furthermore, miR-6838-5p mimics suppressed CDV3 expression in GIST (Fig. 5F).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Shortly, CDV3 is modified by miR-6838-5p.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Fig. 5CDV3 is a target of miR-6838-5p. (
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Bioinformatics website to predict the potential targets of CDV3 and miR-6838-5p, (B and C) Dual luciferase reporter assay and RIP assay to detect the targeting relationship between CDV3 and miR-6838-5p, (D and E) CDV3 expression in tissues and cells, (F) Effect of miR-6838-5p on CDV3 expression; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.Fig 5 CDV3 is a target of miR-6838-5p. (
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Bioinformatics website to predict the potential targets of CDV3 and miR-6838-5p, (B and C) Dual luciferase reporter assay and RIP assay to detect the targeting relationship between CDV3 and miR-6838-5p, (D and E) CDV3 expression in tissues and cells, (F) Effect of miR-6838-5p on CDV3 expression; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
To investigate circAPP/miR-6838-5p/CDV3 affecting GIST biological behavior, this study co-transfected pcDNA 3.1-circAPP and si-CDV3 into GIST-882 cells.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Changes in miR-6838-5p and CDV3 expression were examined.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
pcDNA 3.1-circAPP inhibited miR-6838-5p and elevated CDV3 levels, while si-CDV3 had no effect on miR-6838-5p expression but reversed CDV3 expression (Fig. 6A, B).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
The functional rescue of pcDNA 3.1-circAPP by si-CDV3 was subsequently confirmed in the aspects of clonality, the number of Edu-positive cells, apoptosis rate, cellular glucose consumption, lactate production, ATP levels, ATP/ADP ratio, NAD/NADH ratio, HK2 and PKM2 protein expression (Fig. 6C‒H).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Taken together, circAPP promotes GIST proliferation and the Warburg effect via the miR-6838-5p/CDV3 axis.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Fig. 6CircAPP promotes GIST proliferation and the Warburg effect by regulating the miR-6838-5p/CDV3 axis.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
pcDNA 3.1-circAPP and si-CDV3 were co-transfected, (A) Expression changes of miR-6838-5p, (B) Expression changes of CDV3, (C and D) Colony formation and EdU assays to assess cell proliferation, (E) Flow cytometry to analyze apoptosis, (F) Cellular glucose consumption, lactate production, and ATP levels, (G) ATP/ADP ratio and NAD/NADH ratio, (H) Western blot to evaluate cellular HK2 and PKM2 protein expression; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.Fig 6 CircAPP promotes GIST proliferation and the Warburg effect by regulating the miR-6838-5p/CDV3 axis.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
pcDNA 3.1-circAPP and si-CDV3 were co-transfected, (A) Expression changes of miR-6838-5p, (B) Expression changes of CDV3, (C and D) Colony formation and EdU assays to assess cell proliferation, (E) Flow cytometry to analyze apoptosis, (F) Cellular glucose consumption, lactate production, and ATP levels, (G) ATP/ADP ratio and NAD/NADH ratio, (H) Western blot to evaluate cellular HK2 and PKM2 protein expression; data are expressed as mean ± SD (n = 3). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
This study examined the effect of circAPP on GIST tumor growth and liver metastasis in vivo.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
As demonstrated in Fig. 7A-C, ablating circAPP suppressed GIST tumor volume and weight.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Immunohistochemistry showed that circAPP knockdown suppressed HK2, PKM2, and Ki-67 in GIST tumors (Fig. 7D).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Furthermore, knockdown of circAPP suppressed CDV3 expression in tumors (Fig. 7E).
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Liver metastasis of GIST tumors was assessed by HE-staining (Fig. 7F), manifesting that circAPP depletion suppressed the number of metastatic nodules in the liver.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
circAPP mediates GIST tumor growth and metastasis.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Fig. 7CircAPP inhibits GIST tumor growth and metastasis in vivo. (
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Representative image of tumors, (B) Tumor volume, (C) Tumor weight, (D) IHC image of HK2, PKM2, and Ki-67 proteins in tumors, (E) Western blot detection of tumor CDV3 expression, (F) HE images of tumor liver metastases; data are expressed as mean ± SD (n = 6). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.Fig 7 CircAPP inhibits GIST tumor growth and metastasis in vivo. (
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Representative image of tumors, (B) Tumor volume, (C) Tumor weight, (D) IHC image of HK2, PKM2, and Ki-67 proteins in tumors, (E) Western blot detection of tumor CDV3 expression, (F) HE images of tumor liver metastases; data are expressed as mean ± SD (n = 6). *
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
p < 0.05.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
GIST accounts for 1 %‒3 % of gastrointestinal tumors.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
CircRNAs are imperative regulators in various physiological processes and the occurrence of GIST.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
This study found that circAPP expression was augmented in GIST, and knocking down circAPP hampered GIST cell proliferation and the Warburg effect, triggered cell apoptosis, and limited GIST tumor growth and liver metastasis.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
CircRNAs have become a research hotspot in recent years.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
As research progresses, the functions and mechanistic network of circRNAs in cancer have been implicated in tumor cell activities, including Warburg effect.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
The Warburg effect, proposed by Dr. Otto Heinrich Warburg in 1920, refers to the shift of tumor cell metabolism from oxidative phosphorylation to aerobic glycolysis induced by mitochondrial respiration damage.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
The Warburg effect is a central contributor to the mechanisms of cancer progression, contributing to the cellular progression of cancer cells and being involved in immune responses and drug resistance.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Therefore, inhibiting the Warburg effect inhibits cancer progression.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
This study confirms that circAPP was a positive regulator of the Warburg effect.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Similar to a previous report, this study also presented that circAPP was overexpressed in GIST.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
The in vitro results reported that downregulating circAPP inhibited GIST cell proliferation and the Warburg effect.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Distant metastasis has been reported to be obstructive to cancer treatment.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
Therefore, this study further verified the suppressive impacts of silencing circAPP on GIST tumor growth and liver metastasis in vivo.
PMC11269792
CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor
miRNA sponging is the most well-studied mechanism of circRNA action, and circRNA competitively binds miRNA through complementary base pairing to inhibit the binding of miRNA to its target molecule, thereby regulating the expression of target Mrna.