Effect of miR-4728-3p on the Regulation of PTEN in Apatinib-resistance SCLC Cell Line
Effect of miR-4728-3p on the Regulation of PTEN in Apatinib-resistance SCLC Cell Line
Fang Chen
ABSTRACT
To explore the effects of microRNA (MicroRNA, miR)-4728-3p targeting regulation of phosphatase and tensin homology deleted on chromosome 10 (PTEN) on the drug resistance of small cell lung cancer (SCLC) to apatinib. Human SCLC cell line H69 was purchased from the American Type Culture Collection. Through stepwise screening, an apatinib-resistant SCLC cell line (H69/Apatinib) was established. All cell lines were stored in RPMI-1640 medium, which was supplemented with 10% fetal bovine serum, 100 µg/ml streptomycin and 100 U/ml penicillin and kept in humid air containing 5% CO2 at 37°C. The cell assay was performed when the cells were in the logarithmic growth phase. In the cultured H69/apatinib, the IC50 of apatinib was greater than 30nM, while it was about 13μM in H69 parental cells (P<0.05), indicating that the anti-apatinib SCLC cells were successfully constructed in vitro. The expression of miR-4728-3p mRNA in the miR-4728-3p inhibitor group decreased, and the expression of PTEN mRNA increased (P<0.05). The miR-4728-3p mimic group showed no significant effect on the activity of mutant 3’UTR luciferase of PTEN (P>0.05). The miR-4728-3p inhibitor group had less cell migration and invasion, and its apoptosis rate increased (P<0.05). The abnormal expression of miR-4728-3p mediated the apatinib resistance of SCLC by targeting PTEN and regulating PI3 K/AKT pathway.
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