索拉菲尼对EGFR及K-ras基因突变肺癌细胞株的增殖抑制作用及其作用机制

Inhibitory effect of sorafenib on human none-small cell lung cancer cells harboring EGFR gene mutation or KRAS gene mutation and its mechanism

目的探讨索拉菲尼对非小细胞肺癌PC9细胞株(EGFR基因突变)及A549细胞株(K-ras基因突变)的抑制作用,并探讨其作用机制。方法噻唑蓝(MTT)法检测索拉菲尼分别对PC9及A549细胞的增殖抑制作用。流式细胞仪检测对细胞周期的影响。Western blot法检测p-ERK及ERK表达水平的影响。结果在0.78～25μmol/L浓度范围内,索拉菲尼明显抑制PC9及A549细胞的增殖,且具有时间和浓度的依赖性。流式细胞仪细胞周期分析结果显示,索拉菲尼将PC9和A549细胞阻滞于G1期。Western blot结果显示,索拉菲尼能够下调A549细胞p-ERK的表达水平。结论索拉菲尼能抑制表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)敏感及耐药肺癌细胞株PC9及A549的生长,推测索拉菲尼抗增殖机制可能与对细胞周期的影响以及抑制Ras-Raf-MEK-ERK生存通路有关。

Objective To research the inhibitory effects and mechanism of sorafenib on epidermal growth factor receptor tyrosine kinase inhibitors（ EGFR TKIs）-sensitive human non-small cell lung cancer（ NSCLC ） cell line PC9 and EGFR TKIs-resistant NSCLC cell line A549 in vitro. Methods The inhibitory effects on PC9 and A549 cells exposed to sorafenib were evaluated by MTT method. Cell cycle was detected by flow cytometry. The p-ERK and ERK protein expression levels were measured by Western blot. Results The MTT assay showed sorafenib, dose dependent and time-dependent, had significantly anti-proliferative effects at the concentration ranged from 0.78 to 25 μmol/L. Flow cytometry assay showed that cell cycle was arrested at the G1 phase on PC9 and A549 cells. Western blot assay indicated that sorafenib down-regulated the protein level of p-ERK. Conclusion Sorafenib can inhibit the growth of EGFR TKIs-sensitive PC9 cells and EGFR TKIs-resistant A549 cells. The mechanism may be related to the changes of cell cycle and the inhibition of Ras-Raf-MEK-ERK survival pathway.