特异性下调Grp78表达增加肝癌细胞对厄洛替尼的敏感性  

Knockout Grp78 expression increases the sensitivity of hepatoma cells to erlotinib

作　　者：宋佳[1] 李鑫[2] 李丹[2] 叶丽平[1] 

机构地区：[1]辽宁医学院病理生理学教研室,辽宁锦州121001 [2]辽宁医学院科学实验中心,辽宁锦州121001

出　　处：《山东大学学报（医学版）》2015年第6期7-12,27,共7页Journal of Shandong University:Health Sciences

基　　金：辽宁省博士启动基金(20101065)

摘　　要：目的探讨特异性下调Grp78表达后肝癌细胞对厄洛替尼敏感性的变化及其机制。方法通过用si RNA技术特异性地下调肝癌细胞SMMC-7721中Grp78的表达并做Western blotting鉴定,MTT法检测不同浓度(0、2.5、5、10、20μmol/L)厄洛替尼作用下肝癌细胞存活率的变化,流式细胞术检测细胞凋亡的变化,吖啶橙荧光染色法检测细胞凋亡的形态学变化,免疫印迹法检测AKT、p-AKT、ERK和p-ERK表达。结果转染si RNA-Grp78后24、48、72 h均能明显抑制Grp78的表达,其中48 h抑制效果最明显(P<0.05)。与SMMC-7721/si Control相比,厄洛替尼对SMMC-7721/si RNA-Grp78细胞具有明显的增殖抑制作用和促凋亡作用,差异有统计学意义(P<0.05)。与SM M C-7721/si Control相比,厄洛替尼可明显降低SM M C-7721/si RNA-Grp78细胞中p-ERK和p-AKT的蛋白表达(P<0.05),但总ERK和AKT蛋白表达不受影响。结论特异性下调肝癌细胞中Grp78的表达通过抑制ERK和AKT蛋白的磷酸化增强肝癌细胞对厄洛替尼的敏感性。Objective To explore the effects of knockout Grp78 expression on the sensitivity of SMMC-7721 cells to erlotinib and the molecular mechanism. Methods The Grp78 expression in SMMC-7721 cells was downregulated with si RNA technique and the expression was identified with Western blotting. The survival rates of cells treated with different concentrations of erlotinib（ 0,2. 5,5,10,20 μmol / L） were assessed with MTT method. Cell apoptosis was detected with flowcytometry. The morphological changes of apoptosis were evaluated with acridine orange staining.The expression of AKT,p-AKT,ERK and p-ERK were determined with Western blotting. Results The si RNA transfection significantly inhibited Grp78 expression after 24,48 and 72 hours,and the strongest inhibitive effect appeared after 48 hours（ P 0. 05）. Compared with SMMC-7721 / si Control cells,erlotinib significantly inhibited the proliferation and promoted apoptosis of SMMC-7721 / si RNA-Grp78 cells（ P 0. 05）,and markedly reduced the expression of p-ERK and p-AKT in SMMC-7721 / si RNA-Grp78 cells（ P 0. 05）,while the total protein expression of ERK and AKT were not affected. Conclusion Knockout Grp78 expression increases the sensitivity of hepatoma cells to erlotinib by decreasing the phosphorylation of ERK and AKT.

关 键 词：GRP78 肝癌 厄洛替尼 

分 类 号：R735[医药卫生—肿瘤]