Effect of Lidocaine on Imatinib Resistance in Gastrointestinal Stromal Tumor Cells by Regulating PINK1/Parkin Signaling Pathway
WANG Changyong1, CHEN Weilong1*, LI Longhao2
This study was to explore the effect of LIC (lidocaine) on imatinib resistance in GIST (gastro intestinal stromal tumor) cells by regulating PINK1 (PTEN induced putative kinase 1)/Parkin signaling pathway. GIST cells (GIST-T1) were cultured in vitro and treated with imatinib to induce drug resistant cells. Drug resistant GIST-T1 cells were randomly assigned into NC group (0 μmol/L LIC treatment), LIC low-dose group (500 μmol/L LIC treatment), LIC medium-dose group (750 μmol/L LIC treatment), LIC high-dose group (1 000 μmol/L LIC treat ment), and T0467 group (1 000 μmol/L LIC+5 μmol/L PINK1/Parkin signaling pathway activator T0467). Normal GIST-T1 cells were selected as GIST-T1 group (control group). MTT assay was performed to detect cell viability. Clone formation experiment was performed to measure cell proliferation. Flow cytometry was used to measure cell apoptosis. Scratch experiment and Transwell experiment were used to measure cell migration and invasion. The JC-1 method was used to detect mitochondrial membrane potential. DCFH-DA detection kit was used to detect ROS (reactive oxygen species). Western blot was used to measure autophagy related proteins (LC3Ⅱ/Ⅰ, Beclin-1) and PINK1/Parkin signaling pathway proteins. Compared with GIST-T1 group, there was no significant difference in apoptosis rate, ROS, survival rate, migration rate, invasion number, mitochondrial membrane potential, and the level of LC3Ⅱ/Ⅰ, Beclin-1, PINK1 and Parkin in NC group (P>0.05). Compared with the NC group, the LIC low dose group, LIC medium-dose group, and LIC high-dose group showed an increase in apoptosis rate and ROS (P<0.05), and a decrease in survival rate, migration rate, invasion number, mitochondrial membrane potential, LC3Ⅱ/Ⅰ, Beclin-1, PINK1, and Parkin (P<0.05). Compared with the LIC high-dose group, the T0467 group showed a decrease in apoptosis rate and ROS (P<0.05), and an increase in survival rate, migration rate, invasion number, mitochondrial membrane potential, LC3Ⅱ/Ⅰ, Beclin-1, PINK1, and Parkin (P<0.05). The results of nude mice trans planted tumors showed that the growth rate of nude mice transplanted tumors in the LIC group was slower, the volume and mass of transplanted tumors were reduced, and the expression of LC3 Ⅱ/Ⅰ, Beclin-1, PINK1 and Parkin protein was decreased (P<0.05). LIC can enhance the sensitivity of GIST-T1 cells to imatinib, which may be re lated to the inhibition of PINK1/Parkin signaling pathway.
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