洛伐他汀克服吉非替尼耐药与整合素β1、Survivin的相关性分析

Association of lovastatin overcoming gefitinib resistance with integrin β1 and Survivin

目的研究洛伐他汀克服吉非替尼耐药与整合素β1、Survivin的相关性,并探讨其可能机制。方法将吉非替尼耐药的非小细胞肺癌(NSCLC)细胞株PC9分为4组:无药对照组(RPMI 1640培养液培养)、吉非替尼组(1μmol/L吉非替尼)、洛伐他汀组(5μmol/L洛伐他汀)和吉非替尼与洛伐他汀联合组(1μmol/L吉非替尼+5μmol/L洛伐他汀)。各组细胞分别用相应药物处理后,采用CCK-8法检测细胞的增殖抑制率,PCR法检测细胞中整合素β1、Survivin mRNA的表达水平,蛋白质印迹法检测整合素β1、Survivin蛋白的表达水平。结果与无药对照组、洛伐他汀组及吉非替尼组相比,洛伐他汀与吉非替尼联合组对吉非替尼耐药PC9细胞增殖的抑制作用增强(P<0.01),细胞中整合素β1、Survivin mRNA及蛋白的表达水平降低(P<0.01)。结论洛伐他汀联合吉非替尼克服吉非替尼耐药是通过阻断整合素β1-p-Akt-Survivin信号通路来实现的,有望成为克服吉非替尼耐药的一种有效策略。

Objective To investigate the association of lovastatin overcoming gefitinib resistance with the levels of integrin β1 and Survivin in human non-small cell lung cancer （NSCLC） cell line PC9 in vitro, and to explore the possible mechanism. Methods The NSCLC cell line PC9 with acquired gefitinib-resistance were divided into 4 groups： control group （RPMI 1640）, gefitinib group （1 μmol/L gefitinib）, lovastatin group （5 μlmol/L lovastatin）, and gefitinib combined with lovastatin group （1μmol/L gefitinib＋5 μmol/L lovastatin）. After treatment with different drugs in each group, the inhibition of cell proliferation was detected by cell counting kit-8 （CCK-8） test, the levels of integrin β1 and Survivin mRNA were detected by PCR, and the expressions of integrin β1 and Survivin protein were detected by Western blotting analysis. Results Compared with the control group, lovastatin group and gefitinib group, lovastatin combined with gefitinib treatment had significantly inhibited proliferation of PC9 cells with acquired gefitinib-resistance （P〈0. 01）, and the expressions of integrin β1, Survivin protein and mRNA in PC9 cells were significantly decreased in the three groups （P〈0. 05, P〈0. 01）. Conclusion Mechanisms of lovastatin combined with gefitinib in overcoming gefitinib resistance may be through blocking integrin β1-p-Akt-Survivin signaling pathway, indicating that the combination treatment might be an effective strategy for gefitinib resistance.