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目的探讨Yes相关蛋白1(YAP1)基因在肺腺癌耐厄洛替尼(ER)中的作用及可能机制。方法在PC-9细胞与耐ER PC-9细胞(PC-9/ER)中,实时荧光定量PCR(RT-q PCR)检测YAP1 mRNA表达;Western印迹与间接免疫荧光分别检测YAP1蛋白表达与定位。以YAP1抑制剂维替泊芬(VP)作用PC-9/ER细胞后,比较处理前后YAP1 mRNA和蛋白表达改变、Yap1下游AKT、p-AKT分子表达变化、Cell Counting Kit 8(CCK-8)法计算PC-9/ER耐药指数改变。结果 PC-9/ER耐药指数为99.80±25.81;与PC-9细胞相比较,YAP1 mRNA和YAP1蛋白在PC-9/ER中表达增高;VP抑制效率为(50.96±5.86)%,同时下调了下游AKT、p-AKT蛋白的表达;PC-9/ER细胞的ER半数抑制浓度(IC50)由(11.10±2.72)降低为(1.47±0.32)μmol/L(P=0.024),耐药指数降为原来的1/8。结论 YAP1介导耐ER肺腺癌的作用机制可能是通过PI3K/AKT信号通路,YAP1可能成为肺癌治疗的靶标。
Objective To investigate the role and possible mechanism of Yes-related protein 1 (YAP1) gene in erlotinib-resistant lung adenocarcinoma (ER). Methods YAP1 mRNA expression was detected by real-time quantitative PCR (RT-q PCR) in PC-9 cells and ER-9 / ER cells. Western blot and indirect immunofluorescence were used to detect the expression and localization of YAP1 protein . The effect of YAP1 inhibitor VP1 on the expression of YAP1 mRNA and protein, the expression of AKT and p-AKT downstream of Yap1, Cell Counting Kit 8 (CCK-8) Calculated PC-9 / ER resistance index changes. Results The resistance index of PC-9 / ER was 99.80 ± 25.81. Compared with PC-9 cells, the expression of YAP1 mRNA and YAP1 protein was increased in PC-9 / ER and the inhibitory rate of VP was (50.96 ± 5.86)% (IC50) of PC-9 / ER cells decreased from (11.10 ± 2.72) to (1.47 ± 0.32) μmol / L (P = 0.024) Down to the original 1/8. Conclusion The mechanism of YAP1-mediated ER-resistant adenocarcinoma may be through the PI3K / AKT signaling pathway, and YAP1 may be the target of lung cancer therapy.