Noxa在胃腺癌细胞耐受内质网应激诱导凋亡中的作用  

The role of Noxa in endoplasmic reticulum stress-induced apoptosis in gastric adenocarcinoma cell

作　　者：王家钰[1] 张丽[1] 吴萍[1] 朱孝峰[2] 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 [2]中山大学肿瘤防治中心,广州510060

出　　处：《安徽医科大学学报》2013年第6期590-595,共6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);华南肿瘤学国家重点实验室开放项目

摘　　要：目的研究胃腺癌细胞对内质网应激诱导细胞凋亡耐受中BH3-only家族蛋白Noxa的表达、作用以及相关机制。方法衣霉素(TM)处理SGC-7901中分化胃腺癌细胞株及BGC-823低分化胃腺癌细胞株;采用PI单染法测定细胞凋亡率;Western blot法检测葡萄糖调节蛋白78(GRP78)、半胱天冬氨酸蛋白酶3(Caspase-3)、半胱天冬氨酸蛋白酶9(Caspase-9)、成人T细胞白血病诱导的PMA反应蛋白(Noxa)和髓样细胞白血病-1(Mcl-1)蛋白变化;小干扰RNA(siRNA)技术特异"沉默"Mcl-1和Noxa基因;实时荧光定量PCR检测Noxa mRNA水平变化。结果 TM处理胃腺癌细胞后,GRP78表达升高;胃腺癌细胞对TM诱导的内质网应激性凋亡耐受,Caspase-3和Caspase-9活化较弱;Noxa在蛋白水平和mRNA水平表达均升高;Mcl-1在蛋白水平表达升高;在SGC-7901细胞中,"沉默"Mcl-1基因会使细胞凋亡率升高,而双"沉默"Mcl-1和Noxa基因则细胞凋亡率明显下降。结论胃腺癌SGC-7901细胞在内质网应激状态下,Noxa的促凋亡作用被同样上调的Mcl-1抗凋亡作用拮抗,是胃腺癌细胞耐受内质网应激诱导凋亡的重要机制。Objective To investigate the role and mechanism of Noxa in resistance of gastric adenocarcinoma cells to endoplasmic reticulum （ER） stress-induced apoptosis. Methods Apoptosis was measured by flow cytometry using propidium iodide staining. The activation of Caspase-3, Caspase-9 and protein levels of GRP78, Noxa, Mcl-1 were detected by Western blot. The expression of Noxa mRNA was monitored by quantitative RT-PCR. Silencing of gene Mcl-1, Noxa expression was accomplished by small interfering RNA （siRNA） techniques. Results Gastric adenocarcinoma cells were generally resistant to apoptosis induced by the ER-stress inducer tunicymicin（TM）. Exposure of TM resulted in less activation of Caspase-3 and Caspase-9 in gastric adenocarcinoma cells. The expression of Noxa and Mcl-1 were increased in protein level. Inhibiting Mcl-1 expression by siRNA in SGC-7901 cells increased the cell apoptosis. In contrast, inhibition of Noxa and Mcl-1 partially blocked tunicamycin-induced apoptosis in SGC-7901 cells. Conclusion Pro-apoptotic Bcl-2 family member Noxa is critical in gastric adenocarcinoma cells and the role of pro-apoptotic of Noxa is neutralized by the anti-apoptotic protein Mcl-1, which is also increased by ER stress.

关 键 词：胃肿瘤 内质网应激 NOXA MCL-1 小干扰RNA 

分 类 号：R735.2[医药卫生—肿瘤] R329.25[医药卫生—临床医学]