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山东大学学报（医学版） ›› 2015, Vol. 53 ›› Issue (6): 7-12.doi: 10.6040/j.issn.1671-7554.0.2014.811

• 基础医学 • 上一篇    下一篇

特异性下调Grp78表达增加肝癌细胞对厄洛替尼的敏感性

宋佳1, 李鑫2, 李丹2, 叶丽平1   

  1. 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
  • 收稿日期:2014-11-11 修回日期:2015-03-20 出版日期:2015-06-10 发布日期:2015-06-10
  • 通讯作者: 叶丽平。 E-mail:ye960519@126.com E-mail:ye960519@126.com
  • 基金资助:
    辽宁省博士启动基金(20101065)

Knockout Grp78 expression increases the sensitivity of hepatoma cells to erlotinib

SONG Jia1, LI Xin2, LI Dan2, YE Liping1   

  1. 1. Department of Pathophysiology, Liaoning Medical University, Jinzhou 121001, Liaoning, China;
    2. Science Laboratory Center, Liaoning Medical University, Jinzhou 121001, Liaoning, China
  • Received:2014-11-11 Revised:2015-03-20 Online:2015-06-10 Published:2015-06-10

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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siRNA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、20 μmol/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siRNA- 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/siControl相比,厄洛替尼对SMMC-7721/siRNA-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SMMC-7721/siControl相比,厄洛替尼可明显降低SMMC-7721/siRNA-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。

关键词: 肝癌, Grp78, 厄洛替尼

Abstract: 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 siRNA 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 flow cytometry. 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 siRNA 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/siControl cells, erlotinib significantly inhibited the proliferation and promoted apoptosis of SMMC-7721/siRNA-Grp78 cells (P<0.05), and markedly reduced the expression of p-ERK and p-AKT in SMMC-7721/siRNA-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.

Key words: Erlotinib, Grp78, Hepatocellular carcinoma

中图分类号: 

  • R735
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