E1A基因对三阴乳腺癌细胞侵袭迁移的影响及机制  

Effects of E1A gene on migration and invasion of triple-negative breast cancer cells

作　　者：邰亦成[1] 方琦[1] 谈珂岚[1] 黄贵[1] 张凌焱 

机构地区：[1]常州市第一人民医院,江苏常州213000 [2]杭州市下沙医院妇产生殖中心

出　　处：《山东医药》2017年第29期23-25,共3页Shandong Medical Journal

基　　金：江苏省科学技术厅科技成果(20150014);常州科技计划项目(CJ20140028)

摘　　要：目的探讨E1A基因对三阴乳腺癌(TNBC)细胞侵袭迁移的影响及机制。方法采用脂质体转染法将E1A真核表达载体(pcDNA3.0-E1A)(E1A稳转克隆组)、pcDNA3.0空载质粒(空载克隆组)转染至TNBC MDAMB-231细胞中,G418筛选得到E1A稳定过量表达的细胞克隆,以未转染质粒的MDA-MB-231细胞为空白对照组。采用Western blotting法对稳转克隆进行鉴定。细胞划痕试验检测各组细胞的迁移能力,Transwell小室试验检测各组细胞的侵袭能力,Western blotting法检测各组细胞中HSPA5蛋白表达水平。结果 G418筛选建立稳定转染pcDNA3.0-E1A质粒的MDA-MB-231细胞克隆(E1A稳转克隆)。与空白对照组和空载克隆组相比,E1A稳转克隆组的细胞迁移和侵袭能力均降低(P均<0.05),细胞HSPA5蛋白表达水平下降(P均<0.05)。结论 E1A基因能够明显抑制TNBC细胞的侵袭迁移,其机制可能与EA1促进HSPA5蛋白降解有关。Objective To investigate the effects of E1A gene on the migration and invasion of triple-negative breast cancer（ TNBC） cells and is mechanism. Methods The eukaryotic expressive vector containing E1A gene（ pcDNA3. 0-E1A）（ E1A stable clone group） and pcDNA3. 0 empty plasmid（ empty-plasmid clone group） were separately transfected into MDA-MB-231 cells by the lipofectamine,and stable E1A positive clones were selected by G418. Meanwhile,the MDA-MB-231 cells without any transfection were taken as the blank control group. Then Western blotting was utilized to identify the stable clones. The migration and invasion abilities of cells in all groups were analyzed by Scratch test and Transwell assay,respectively. Besides,the protein level of HSPA5 in all groups was detected by Western blotting. Results MDA-MB-231 cell clones stably transfected with pcDNA3. 0-E1A plasmid（ E1A clones） were successfully established by the G418. The cell migration and invasion abilities of E1A clones in the E1A stable clone group were much lower as compared with those of the blank control group and the empty-plasmid clone group（ P 0. 05）,accompanied by the down-regulated expression of HSPA5 protein（ all P 0. 05）. Conclusion E1A gene can significantly inhibit the cell invasion and migration of TNBC cells in vitro by E1A promoting the protein degradation of HSPA5.

关 键 词：三阴乳腺癌 E1A 细胞侵袭 细胞迁移 

分 类 号：R737.9[医药卫生—肿瘤]