阿托伐他汀通过抑制RhoA活性减少非小细胞肺癌的增殖

Atorvastatin reduces proliferation of non-small cell lung carcinomas via inhibition of RhoA activity

目的观察阿托伐他汀(ATV)对非小细胞肺癌(NSCLCs)细胞增殖的影响并探讨其机制。方法用改进MTT法检测不同浓度ATV、RhoA特异性抑制剂C3转移酶(C3 transferase)、RhoA siRNA处理后A549细胞、H358细胞、Calu3细胞的增殖情况。用RhoA活性测定法观察C3 transferase及不同浓度、不同时间ATV处理后NSCLCs细胞的RhoA活性变化。用Lipofectamine2000转染RhoA siRNA,Western blot检测RhoA的蛋白表达。结果 ATV浓度依赖性(0.1～25.0μmol/L)地抑制三株NSCLCs细胞增殖(P<0.05或P<0.01)。ATV浓度依赖性(0.1～25.0μmol/L)和时间依赖性(12～96 h、48～96 h)地抑制不同NSCLCs细胞的RhoA活性(P<0.05或P<0.01)。C3 transferase能显著降低三株NSCLCs细胞的RhoA活性并抑制NSCLCs细胞增殖(P<0.01)。RhoA siRNA在有效沉默RhoA蛋白表达后,也明显抑制NSCLCs细胞增殖(P<0.01)。但ATV联合应用C3 transferase或RhoA siR-NA不能产生协同抑制效应,而与单用C3 transferase或RhoA siRNA的抑制增殖作用相似(P>0.05)。结论 ATV主要通过抑制RhoA活性从而抑制NSCLCs细胞增殖。

Objective To evaluate the effect of atorvastatin （ATV） on proliferation of non-small cell lung carcinomas（NSCLCs） as well as the underlying molecular mechanisms. Methods Cell proliferation was measured by a modified MTT assay after the treatment with different concentrations of ATV, RhoA specific inhibitor C3 transferase and RhoA siRNA in A549, H358, and Calu3 cells. The change of RhoA activity was measured by a microplate reader in NSCLCs cells treated with C3 transferase or atorvastatin at different doses and different times. Transfection of NSCLCs with RhoA siRNA was done using Lipofectamine 2000. Expression levels of RhoA protein were determined by Western blot. Results Atorvastatin significantly reduced the preliferation of three NSCLCs cell lines in concentration-dependent manner （0.1 25.0 panol/L） （P 〈0.05 or P 〈0.01 ）. Atorvastatin substantially inhibited the RhoA activity of different NSCLCs cell lines in a concentration-dependent （0.1 ～ 25.0 μmol/L） and time-dependent manner （ 12 ～96 hours, and 48 ～ 96 hours） （P 〈 0.05 or P 〈 0.01 ）. C3 transferase significantly inhibited the RhoA activity of the three NSCLCs cell lines and resultantly inhibited NSCLCs cells proliferation （P 〈 0.01 ）. Moreover,knockdown of RhoA expression with RhoA siRNA also resulted in a significant inhibition of NSCLCs cells proliferation （P 〈 0.01 ）. However, additional atorvastatin administration exerted no additional growth inhibition in C3 transferase-treated or RhoA siRNA-treated NSCLC cells compared with the inhibitor or specific siRNA treatment alone （P 〉 0.05）. Condusion Atorvastatin can sup- press NSCLCs cells proliferation primarily through inhibition of RhoA activity.