NVP-BEZ235体外对胆管癌细胞株QBC939增殖及凋亡的影响

Effects of NVP-BEZ235 on proliferation and apoptosis with human cholangiocarcinoma QBC939 cell line in vitro

目的研究NVP-BEZ235体外对胆管癌细胞株QBC939增殖及凋亡的影响。方法在体外培养人胆管癌细胞株QBC939,通过流式细胞仪检测NVP-BEZ235对QBC939凋亡的影响;MTT法检测NVP-BEZ235作用24小时、48小时和72小时对QBC939的增殖影响;Western blot分析NVP-BEZ235作用48小时对QBC939凋亡的影响(PARP),并检测分析相关信号通路蛋白Bcl-2、Akt、p-Akt、c-FLIPL和Mcl-1表达水平的变化。结果 NVP-BEZ235能抑制QBC939的增殖,抑制作用具有时间依赖性和浓度依赖性,同时能诱使细胞发生凋亡。NVP-BEZ235导致细胞内抗凋亡蛋白Mcl-1、c-FLIPL和Bcl-2表达下调,并降低p-Akt表达。结论 NVP-BEZ235抑制Akt的磷酸化,NVP-BEZ235通过PI3K/Akt通路下调Bcl-2、Mcl-1和c-FLIPL的表达,从而诱使QBC939出现凋亡,抑制QBC939增殖。

Objective To investigate the effects of NVP-BEZ235 on proliferation and apoptosis of human cholangiocarcinoma（ CCA） cell QBC939 in vitro and to reveal the antineoplastic mechanisms of NVP-BEZ235. Methods Human CCA cell line QBC939 was used in this study. Cell apoptosis by NVPBEZ235 was analyzed using the flow cytometry. Cell growth inhibition by NVP-BEZ235 for 24 h 48h 72 h by MTT assay. The antineoplastic mechanisms of NVP-BEZ235 for 48 h were assessed by western blotting for PARP、Bcl-2、Akt、p-Akt、c-FLIPLand Mcl-1 assay in QBC939 cell. Results NVP-BEZ235 treatment inhibited the proliferation and induced apoptosis of human CCA cell QBC939,which appeared time-dependent and concentration-dependent effects. NVP-BEZ235 reduced protein levels of Mcl-1、c-FLIPLand Bcl-2and downregulate protein level of p-Akt significantly. Conclusion NVP-BEZ235 inhibited the phosphorylation of Akt. NVP-BEZ235 downregulated Bcl-2、c-FLIPL、Mcl-1 protein level via PI3K/AKT signaling pathway to induce apoptosis and inhibit the proliferation of human CCA cell QBC939.