李利波,陈腾祥,潘娅
LI Libo,CHEN Tenxiang,PAN Ya

• 1:贵州医科大学生理学教研室
• 2:贵州医科大学
• 3:贵州省人民医院肿瘤科

摘要(Abstract)：

目的:探讨正常肝细胞和肝癌细胞在诱导发生内质网应激(ERS)前后细胞裂解液中葡萄糖调节蛋白78(GRP78)、蛋白质二硫键(PDI)、的表达水平。方法:取培养获得的第3代正常肝细胞LO-2和肝癌Hep G2与SMMC-7721细胞,采用150 mmol/L无水乙醇处理48 h诱导发生ERS;分别提取3种细胞裂解液中蛋白质,采用Western blot方法检测GRP78和PDI表达水平。结果:在ERS前,GRP78在Hep G2细胞中表达最高,其次为SMMC-7721细胞,在LO2细胞中表达最低,3种细胞GRP78表达水平比较,差异有统计学意义(P<0.001);诱导细胞发生ERS后,Hep G2细胞及LO2细胞GRP78表达均上调,而SMMC-7721细胞GRP78表达下调,分别与相应细胞发生ERS前比较,差异有统计学意义(P<0.05);在ERS前,PDI的表达在SMMC-7721细胞中最高,其次LO2细胞,在Hep G2细胞中表达最低,3种细胞比较,差异有统计学意义(P<0.001);诱导发生ERS后,与ERS前相对应细胞比较,3种细胞的PDI表达水平均下调,差异有统计学意义(P<0.05)。结论:ERS后,肝癌SMMC-7721和Hep G2细胞裂解液中的GRP78表达水平发生变化,PDI表达降低。
Objective: To investigate the expression level of PDI and GRP78 before and after ERS in the cell lysates of normal liver cells and liver cancer cells. Methods: The third-generation cultivated LO-2,Hep G2 and SMMC-7721 cells were processed by 150 mmol / L anhydrous ethanol for 48 h to induce ERS. Then respectively extracted the proteins in three kinds of cell lysis liquid,then adopting Western blot method to detect the expression level of GRP78 and PDI. Results: Before the ERS,the expressing of GRP78 in Hep G2 cells was the highest,secondly was SMMC-7721 cells,the expressing of GRP78 in LO2 cells was the lowest. To compare the expression of GRP78 in three kinds of cells,differences were statistically significant( P < 0. 001). Before ERS,the expression of PDI in SMMC-7721 cells was highest,followed by LO2 cells,the expression in Hep G2 cells was the lowest. To compare the expression of three kinds of cells,differences were statistically significant( P < 0. 01). After induced ERS,all of the expression of PDI in three cells were downregulating,differences were statistically significant( P < 0. 05). Conclusion: After ERS,the expression levels of GRP78 changed in SMMC-7721 and Hep G2 cells lysis liquid,expression level of PDI decreased.

关键词(KeyWords)： 癌,肝细胞;内质网应激;蛋白质二硫键异构酶;葡萄糖调节蛋白78
carcinoma hepatocellular;endoplasmic reticulum stress;protein disulfide isomerase;glucose regulated protein 78

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技厅科研基金黔科合[LG(2011)208号];; 贵州省教育厅自然科学研究基金黔教科[(2008)022号]

作者(Author): 李利波,陈腾祥,潘娅
LI Libo,CHEN Tenxiang,PAN Ya

DOI: 10.19367/j.cnki.1000-2707.2016.06.011

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