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PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The mice acclimated in the animal room for 6 days before the experiment.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
All the mice were transformed into subcutaneous xenograft models.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Cells of normal or drug-resistant strains of GIST882 in the logarithmic growth stage were collected, prepared into a single-cell suspension, adjusted to 1×10/mL, mixed with Matrigel gel (10 mg/mL, BD Biosciences) of equal proportion.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Human fibroblasts in the ratio 1:10,000 were added, mixed, and set aside.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The injected volume was 0.1 mL/animal, and the number of cells in normal or drug-resistant strains containing GIST882 was 5×10/animal.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
After the injection, there was a protruding cutaneous mound in the right axilla.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The tumor-bearing mice were randomly divided into 5 groups according to the tumor volume when the tumor grew to 100 mm, i.e., the normal group, imatinib-resistant group, imatinib group, CQ group, and imatinib + CQ group.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Five mice were found in each group.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
For the imatinib group, imatinib mesylate capsule powder was dissolved in a 15 mg/mL working solution with normal saline.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The whole process was done in the dark.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The dosage was 150 mg/kg, twice a day for 28 days, and administered by gavage.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
For the CQ group, CQ was dissolved in deionized water to give a 6 mg/mL working solution.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
This was done and stored in the dark.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
It was injected intraperitoneally at a dose of 60 mg/kg once a day for 28 days.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
For the imatinib + CQ group, the dosage of imatinib was 150 mg/kg while that of CQ was 60 mg/kg administered for 28 days.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The normal and imatinib-resistant groups were given the same volume of normal saline for 28 days.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
All experiments involving cell lines and animals were approved by the Hangzhou cancer hospital ethical review board.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
All experiments were performed following the Hangzhou cancer hospital and national guidelines and regulations.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The cell-counting kit (CCK)-8 method and flow cytometry were used for in vitro drug sensitivity testing and autophagy level detection.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Detection of the apoptosis level was by flow cytometry with the annexin V Kit.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Western blotting was used to analyze the role of autophagy and apoptosis in GIST cells with CQ alone, imatinib alone, or in combination, and to analyze MAPK pathway expression.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
In vitro results were confirmed by in vivo experiments using the mice model.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The transplantation model of human GIST resistant to imatinib in nude mice was identified and established.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The above-mentioned imatinib-resistant cell lines and GIST882 were inoculated into nude mice to observe their tumorigenicity.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The tumor size, growth rate, and survival time of nude mice were observed.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Hematoxylin and eosin and immunohistochemical staining were used to detect the pathological characteristics and immunophenotype of the transplanted tumor.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Detection of KIT and PDGFRA gene mutations in the transplanted imatinib-resistant GIST was done by denaturing high performance liquid chromatography (DHPLC) and direct sequencing.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
ERK and KIT expression and regulation levels were detected by Western blotting.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The experimental data were expressed as means ± standard deviations ().
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Analysis of variance (ANOVA) and statistical tests were analyzed means of multiple groups in the GraphPad Prism statistical software.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
First, the data were tested for homogeneity of variance.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
If the variance was homogeneous, the overall comparison was made using the one-way ANOVA, and the two-way mean comparison method between multiple dose groups and one control group.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Abnormal or non-uniform variance data were used for statistics in the rank sum test.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
In this study, the drug concentration gradient method was used to establish the imatinib-resistant GIST (GIST882-ir cell line) from the imatinib-sensitive GIST882 cell line.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The half maximal inhibitory concentration (IC50) was measured using the CCK-8 method to judge the sensitivity/resistance of the cell lines to imatinib.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The results showed growth of the GIST882 cells was affected to a certain extent under the action of imatinib at a low concentration (0.05 μm), and was significantly affected when it was > 0.1 μm.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
However, GIST-ir cells had a strong resistance to imatinib.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
At imatinib concentrations of 0.05–5 μm, there was no significant effect on the growth of GIST-ir cells (Figure 1A and B).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
GIST-ir cells had an inconsistent morphology; some cells were epithelioid, while others were short fusiform or irregularly shaped.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Secondary mutations of KIT/PDGFRA in GIST cells were the most important contributors to imatinib resistance in GISTs.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
In GIST-ir cell lines, KIT sequencing results showed no secondary drug resistance mutation.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Meanwhile, no mutation was found in PDGFRA sequencing.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Flow cytometry was used to detect apoptosis in four quadrants: Q1-UL (necrotic cells and fragments), Q1-UR (late apoptotic and necrotic cells), Q1-LL (normal cells), and Q1-LR (early apoptotic cells, which could be detected by CCK8).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The sum of the Q1-UR and Q1-LR quadrants was used to calculate the apoptosis rate.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The detailed results are shown in Figure 1C.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
After 48 hours, the apoptosis rates of the GIST882 group, the GIST882-ir group, the GIST882-ir+ im group, the GIST882-ir + CQ group, the GIST882-ir +im+ CQ group were 6.4%, 4.3%, 8.6%, 9.2% and 19.3%, respectively.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Imatinib had a weak inhibitory effect on the growth of GIST882-ir cells, while CQ had no effect on the growth of GIST882-ir cells.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The combination of CQ and imatinib greatly enhanced the inhibitory effect of imatinib on the growth of GIST882-ir cells (Figure 1D).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
KIT expression in the GIST882 imatinib-resistant group was significantly higher than that in the GIST882 group (* p<0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the GIST882 imatinib-resistant group, the KIT expression level in the imatinib group and the combination group was significantly lower (p<0.05), but was not significantly different in the CQ group (p>0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
KIT expression in the combination group decreased, though it was not significantly lower than in the imatinib group (p>0.05) (Figure 1F).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The KIT phosphorylation level (p-kit/Kit) in the GIST882 imatinib-resistant group was lower than in the GIST882 group, but there was no significant difference (p>0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the GIST882 imatinib-resistant group, the phosphorylation level in the imatinib group and the combination group was significantly lower (p<0.05), and the phosphorylation level in the CQ group was significantly higher (p<0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The phosphorylation level in the combination group was higher than that in the imatinib group, but there was no significant change (p>0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
ERK expression in the GIST882 imatinib-resistant group was higher than in the GIST882 group, but there was no significant difference (p>0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the, ERK expression in the imatinib group, CQ group, and combination group decreased, but was not significantly different with the GIST882 imatinib-resistant group (p>0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the GIST882 imatinib-resistant group, the ERK phosphorylation (p-ERK/ERK) level in the imatinib group was lower, but not significantly different (p>0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The ERK phosphorylation level of ERK protein in the combination group was lower than that in the imatinib group, but the difference was not significant (p>0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
There was no significant difference in the tumor volume between the drug-resistant group and the normal group.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Three days after administration, tumor volume of the imatinib group and the imatinib + CQ group was non-significantly lower than in the drug-resistant group (Figure 2A).Figure 2(A) Effect on tumor volume. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The immunohistochemical detection of Beclin 1 protein expression in rat tumor tissue. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The immunohistochemical detection of caspase 3 protein expression in rat tumor tissue. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
immunohistochemical detection of caspase 7 protein expression in rat tumor tissue.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Abbreviations: CK, normal control group; im-r, imatinib-resistant group; im, imatinib-resistant cells + imatinib; CQ, imatinib-resistant cells + chloroquine; im+ CQ, imatinib-resistant cells + imatinib + chloroquine. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Effect on tumor volume. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The immunohistochemical detection of Beclin 1 protein expression in rat tumor tissue. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The immunohistochemical detection of caspase 3 protein expression in rat tumor tissue. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
immunohistochemical detection of caspase 7 protein expression in rat tumor tissue.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Abbreviations: CK, normal control group; im-r, imatinib-resistant group; im, imatinib-resistant cells + imatinib; CQ, imatinib-resistant cells + chloroquine; im+ CQ, imatinib-resistant cells + imatinib + chloroquine.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the normal group, there was a significant decrease in the body weight of mice in the resistant group on day 0, day 3, and day 6 (p<0.05, p<0.01).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the resistant group, there was a significant increase in the body weight of the mice in the imatinib group and the CQ group day 0 and day 3 of the drug administration (p<0.01).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The body weight of the mice in the combined group increased significantly on day 0 of drug administration (p<0.01) (Figure 3A).Figure 3(A) Changes of body weight in mice. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Beclin1 expression in tumor tissue detected by immunohistochemistry. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Caspase-3 expression in tumor tissue detected by immunohistochemistry. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Caspase-7 expression in tumor tissue detected by immunohistochemistry. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Changes of body weight in mice. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Beclin1 expression in tumor tissue detected by immunohistochemistry. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Caspase-3 expression in tumor tissue detected by immunohistochemistry. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Caspase-7 expression in tumor tissue detected by immunohistochemistry.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Autophagy level (LC3II/LC3I) analysis showed that the autophagy level in the imatinib-resistant group was lower than that in the normal group, but there was no significant difference (p>0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the imatinib-resistant group, there was an upward trend but no significant change (p>0.05) in the imatinib group and imatinib + CQ group, but there was a downward trend without any significant difference in the CQ group (p>0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the imatinib group, there was an upward trend in the imatinib + CQ group, but no significant difference (p>0.05) (Figure 4A).Figure 4(A) LC3II/I protein expression.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the normal group (CK group).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Imatinib-r group: imatinib-resistant group.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Imatinib-r group vs. CK group, *p<0.05, **p<0.01; each drug administration group vs. Imatinib-r group, #p<0.05, ##p<0.01; Imatinib+CQ group vs. Imatinib group, and p<0.05, and p<0.01. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Analysis of KIT and ERK protein expression and phosphorylation level. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
LC3II/I protein expression.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the normal group (CK group).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Imatinib-r group: imatinib-resistant group.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Imatinib-r group vs. CK group, *p<0.05, **p<0.01; each drug administration group vs. Imatinib-r group, #p<0.05, ##p<0.01; Imatinib+CQ group vs. Imatinib group, and p<0.05, and p<0.01. (
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Analysis of KIT and ERK protein expression and phosphorylation level.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
According to the positive immunohistochemistry (IHC) results, Beclin 1 expression was highest in the imatinib group, followed by the imatinib-resistant group, and the combination group.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The expression in the normal group was the same as that in the CQ group, and lower than that in the other three groups.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Due to the small sample size (n=5) of each group, the standard deviation between groups was large, and there was no statistical significance (Figures 3B and 2B).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the normal group, the expression of Caspase-3 was highest in the resistant cell group, and there was no significant difference in the other groups (Figures 3C and 2C).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The expression of caspase-7 in the drug-resistant group was lower than in the normal cell group, but there was no significant difference between the two groups (Figures 3D and 2D).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
However, the ratio of caspase 3 and 7 protein in the combined group was highest, indicating that apoptosis level was highest in this group.
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
The KIT expression level in the imatinib-resistant group was significantly higher than in the normal group (*p<0.05).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
Compared with the imatinib-resistant group, KIT expression in the imatinib group, CQ group, and combination group were significantly lower (##p<0.01).
PMC7342409
Chloroquine Combined with Imatinib Overcomes Imatinib Resistance in Gastrointestinal Stromal Tumors by Inhibiting Autophagy via the MAPK/ERK Pathway
KIT expression in the combination group was significantly lower than in the imatinib group (andp<0.05) (Figure 4B).