摘要
目的探讨Bcl-2蛋白家族中Mcl-1在衣霉素诱导胃腺癌细胞内质网应激性凋亡中的作用。方法采用流式细胞仪PI单染法检测细胞凋亡率;Westernblot检测Bcl-2、Mcl-1、Bcl-XL在衣霉素诱导胃腺癌细胞内质网应激状态下的表达;脂质体LipofectamineTM2000转染pcDNA3.0/Mcl-1质粒;Westernblot分别检测转染前后Mcl-1蛋白的表达水平变化。结果衣霉素诱导胃腺癌细胞凋亡具有剂量和时间依赖性,但两株细胞对衣霉素诱导的凋亡都不敏感,其中SGC-7901较BGC-823对衣霉素的耐受表现更明显。由于衣霉素诱导SGC-7901和BGC-823的凋亡与线粒体凋亡途径有关,检测Bcl-2蛋白家族Mcl-1在SGC-7901中有明显上调。通过转染质粒高表达Mcl-1蛋白,衣霉素诱导的凋亡有明显下降。结论Mcl-1在衣霉素诱导胃腺癌细胞凋亡中发挥重要的抑制作用。
Objective To study the role of antiapoptosis protein Mcl-1 in the pathway of apoptosis of ER stress-induced gastric adenocarcinoma cells.Methods Apoptosis was measured by flow cytometry using propidium iodide staining.Changes of Bcl-2、Mcl-1、Bcl-XL were detected by Western blot method.The eukaryotic expression vector pcDNA3.0/Mcl-1 containing human Mcl-1 cDNA and vector pcDNA3.0 were transfected into the resistant SGC-7901 cells,respectively.Western blot was used to detect the target gene expression at protein levels.Results ER stress induced apoptosis with tunicamycin of gastric adenocarcinoma cells in a dose-and time-dependent manner.It appeared that SGC-7901 cells could resist to tunicamycin-induced apoptosis more obviously than BGC-823 cells.The expression of Mcl-1 was increased in SGC-7901.Over-expression Mcl-1 by trasfection protects gastric adenocarcinoma cells from tunicamycin-induced apoptosis.Conclusion Antiapoptosis protein Mcl-1 plays an important role in apoptosis induced by the ER stress inducers of gastric adenocarcinoma cell.
出处
《安徽医科大学学报》
CAS
北大核心
2010年第1期20-23,共4页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:30572118)
安徽省自然科学基金(编号:070413077)
关键词
胃肿瘤
细胞凋亡
衣霉素
转染
stomach neoplasms
apoptosis
tunicamycin
transfection