索拉非尼联合紫杉醇体外抗肺癌A549/Taxol细胞作用机制研究

Effect of Anti-Cancer Mechanism of Sorafenib Combined with Taxol on A549/Taxol Cell Line of Lung Cancer in Vitro

目的探讨索拉非尼联合紫杉醇体外对肺癌A549/Taxol细胞的作用机制。方法将对数生长期的A549/Taxol细胞进行分组,空白对照组和药物处理组,药物处理组包括紫杉醇组、索拉非尼组、紫杉醇联合索拉非尼组,各组分别加入不同浓度的相应药物。利用MTT法和流式细胞仪检测不同浓度紫杉醇及其联合索拉非尼对A549/Taxol细胞增殖与凋亡的影响。通过蛋白印迹法、Real-Time PCR观察埃兹蛋白(Ezrin)、磷酸化埃兹蛋白(p-Ezrin)及抗凋亡蛋白(bcl-2)的表达水平。结果与空白对照组比较,紫杉醇组与索拉非尼组分别随着浓度的增加对细胞生长的抑制作用逐渐加大(P<0.05),相对于上述3组,索拉非尼联合紫杉醇组抑制细胞生长更显著(P<0.01);空白对照组A549/Taxol细胞凋亡率仅为6%,紫杉醇组为14%,索拉非尼组为8%,而紫杉醇联合索拉非尼组明显提高为50%,与前3组比较,差异有统计学意义(P<0.01,P<0.05,P<0.01);与空白对照组相比,索拉非尼组、紫杉醇组bcl-2基因表达水平显著下降(P<0.05),而索拉非尼联合紫杉醇组bcl-2表达水平显著低于上述3组(P<0.01);与空白对照组比较,紫杉醇组、索拉非尼组Ezrin、p-Ezrin均有所下降(P<0.05),紫杉醇联合索拉非尼组的Ezrin、p-Ezrin得到明显抑制(P<0.01)。结论索拉非尼联合紫杉醇用药可通过抑制Ezrin及其Bcl-2表达,促进肺癌A549/Taxol细胞凋亡,具有抗肿瘤的作用。

Objective To investigate anticancer mechanism effect of Sorafenib combined with Taxol on A549 /Taxol cell line of lung cancer in vitro. Methods A549 / Taxol cells in logarithmic growth phase were divided into control and medication groups. The medication group included Taxol,Sorafenib and Sorafenib combined with Taxol subgroups,and each subgroup was added with different concentrations of corresponding drugs. Effects of different concentrations of Sorafenib combined with Taxol on cell proliferation and apoptosis of A549 / Taxol cells were detected,using methyl thiazolyl tetrazolium（ MTT） and flow cytometry respectively. Expression levels of Ezrin,p-Ezrin and bcl-2 were detected,using Western Blot and real-time polymerase chain reaction（ PCR） methods. Results Compared with those in control group,inhibition effects on cell proliferation gradually enhanced with increasing concentrations（ P 〈0. 05）,and cell proliferation was significantly inhibited in Sorafenib,Taxol and Sorafenib combined with Taxol groups（ P〈 0. 01）. Apoptosis rates of A549 / Taxol cell line in control,Taxol,Sorafenib and Sorafenib combined with Taxol groups were 6%,14%,8% and 50% respectively,and there were significant differences in the rate in Sorafenib combined with Taxol group compared with those in control,Taxol and Sorafenib groups（ P 〈0. 01,P 〈0. 05,P 〈0. 01）. Compared with those in control group,expression levels of bcl-2 in Taxol and Sorafenib groups were significantly decreased（ P 〈0. 05）,and the level in Sorafenib combined with Taxol group was significantly lower than those in control,Taxol and Sorafenib groups（ P〈 0. 01）; expression levels of Ezrin and p-Ezrin in Taxol and Sorafenib groups were decreased（ P 〈0. 05）,and the expressions in Sorafenib combined with Taxol group were significantly inhibited,and the differences were statistically significant（ P〈 0. 01）. Conclusion Sorafenib combined with Taxol has anticancer effect by inhibiting expressions of Ezrin and bcl-2 and promoting A549 / Taxol apoptosis.