Bcl-2在紫杉醇耐药胃腺癌细胞中的表达及意义  

The role of Bcl-2 in paclitaxel-resistant gastric adenocarcinoma cells

作　　者：张丽[1] 胡汪来 王家钰[1] 张旭东[1] 张林杰[1] 

机构地区：[1]安徽医科大学免疫学教研室,合肥230032

出　　处：《安徽医科大学学报》2014年第8期1048-1052,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省高等学校省级自然科学研究项目(编号:KJ2011A167)

摘　　要：目的探讨凋亡抑制蛋白Bcl-2在紫杉醇(TAX)耐药胃腺癌细胞中的作用。方法采用药物浓度梯度递增持续作用法建立耐TAX SGC-7901细胞株,PI单染法流式细胞仪检测细胞凋亡率差异,MTT比色法检测细胞生存率变化,Western blot检测TAX处理后SGC-7901普通细胞株及耐药细胞株Bcl-2、mcl-1、Bax、Bcl-xl蛋白表达,半定量PCR检测Bcl-2 mRNA水平变化,流式细胞术检测Bcl-2基因沉默后的凋亡率变化,MTT检测细胞生存率。结果 TAX诱导胃腺癌细胞凋亡具有时间和剂量相关性,耐药胃腺癌细胞株对TAX诱导的凋亡不敏感,TAX处理后Western blot检测到耐药细胞株中Bcl-2表达上调,亲本细胞株Bcl-2则表达下调,半定量PCR检测耐药胃腺癌细胞株中Bcl-2 mRNA水平亦上调。在耐药胃癌细胞株中沉默Bcl-2后,Bcl-2蛋白水平下调,细胞凋亡率明显上升,细胞生存率显著下降。结论Bcl-2的表达上调可能是胃腺癌细胞对TAX耐药的重要机制。Objective To investigate the role of Bcl-2 in paclitaxel-resistant gastric adenoearcinoma cells.Methods The paclitaxel-resistant cell line SGC-7901/TAX was established by increasing drug concentration gradient and continuing induction.The difference of apoptosis rate was measured by flow cytometry using propidium iodide staining.MTT colorimetric detected cell survival rate.The protein expression levels of Bcl-2,mcl-1,Bax,Bcl-xl were detected by Western blot.The expression of Bcl-2 mRNA level in the resistant cell line was monitored by RT-PCR.Silencing of gene Bcl-2 expression was accomplished by small interfering RNA techniques.Results Paclitaxel-induced apoptosis in gastric adenocarcinoma cells was in a time and dose-dependent manner.Drug-resistant gastric adenocarcinoma cell line was not sensitive to paclitaxel induced apoptosis.The up-regulation of Bcl-2 protein level was obvious in the drug-resistant cell line,and Bcl-2 mRNA level was also found to be up regulated by semi-quantitative PCR.Inhibiting Bcl-2 expression by SiRNA in resistant gastric adenocarcinoma cell line increased the cell apoptosis.Conclusion Bcl-2 up regulation plays an important role in gastric adenocarcinoma cells resistant to paclitaxel induced apoptosis.

关 键 词：胃肿瘤 紫杉醇 细胞凋亡 肿瘤耐药 BCL-2 

分 类 号：R735.2[医药卫生—肿瘤] R392.12[医药卫生—临床医学]