葡萄糖调节蛋白78影响乳腺癌细胞株MCF-7的黏附及迁移能力的研究

Glucose regulatory protein 78 affects the adhesion and migration of breast cancer cell line MCF-7 through MMP2 and MMP9

目的探讨乳腺癌细胞株MCF-7的黏附及迁移能力与GRP78的关系及调控机理。方法将pEGFP-N1-GRP78质粒转染MCF-7细胞中从而获得GRP7高表达细胞株(MCF-7/GRP78),空载体质粒(pEGFP-N1)转染的细胞(MCF-7/N1)及无任何处理的细胞(MCF-7/UT)为对照。细胞黏附实验及细胞划痕实验验证不同细胞的黏附及迁移能力。Western Blot检测不同细胞中GRP78、MMP2、MMP9的表达情况。结果Western blot结果显示,与MCF-7/UT细胞相比,MCF-7/GRP78细胞中GRP78表达水平明显增加(P<0.05),而MCF-7/N1细胞中GRP78的表达水平与之比较无差异(P>0.05);MCF-7/GRP78细胞的2 h黏附率显著高于MCF-7/UT组和MCF-7/N1组(P<0.05),而后两者黏附率无差异(P>0.05);细胞划痕试验显示MCF-7/GRP78的迁移率明显高于MCF-7/UT组和MCF-7/N1(P<0.05),而后两者无差异(P>0.05);Western Blot检测MCF-7/GRP78细胞中MMP2、MMP9的表达水平明显高于MCF-7/UT和MCF-7/N1(P<0.05),而MCF-7/UT组和MCF-7/N1组无差异(P>0.05)。结论GRP78可以促进MCF-7细胞的黏附及迁移能力,并且MMP2、MMP9的表达水平与GRP78也呈正相关。

Objective To explore the relationship between the adhesion and migration of breast cancer tumor cells(MCF-7)and Glucose regulation protein-78(GRP78)and its regulatory mechanism.Methods The pEGFP-N1-GRP78 plasmid was transfected into MCF-7 cells to obtain a highly expressed GRP7 cell line(MCF-7/GRP78),and cells transfected with pEGFP-N1(MCF-7/N1)and cells without any treatment(McF-7/UT)were the controls.Cell adhesion test and cell scratch test verified the adhesion and migration ability of different cells.The expressions of GRP78,MMP2 and MMP9 in different cells were detected by Western Blot(WB).Results WB results showed that compared with MCF-7/UT cells,the expression level of GRP78 in MCF-7/GRP78 cells was significantly increased(P<0.05),and there were statistical significantly differences,but there were no statistical significantly differences in adhesion between the latter two(P>0.05).The 2 h adhesion rate of MCF-7/GRP78 cells was,significantly higher than that of MCF-7/UT group and MCF-7/N1 group(P<0.05),the difference was statistically significant,and there were not statistically significant differences(P>0.05).Cell scratch test showed that the mobility of MCF-7/GRP78 significantly higher than that of MCF-7/UT groupand McF-7/N1(P<0.05),with statistically significant differences,but no statistically significant differences(P>0.05).The expression levels of MMP2 and MMP9 in MCF-7/GRP78 cells were significantly higher than that of MCF-7/UT and MCF-7/N1 by WB test(P<0.05),Between McF-7/UT and MCF-7/N1 there were no statistically significant differences(P>0.05).Conclusion GRP78 can promote the adhesion and migration of MCF-7 cells,and the expression levels of MMP2 and MMP9 are positively correlated with GRP78.