BRMS1对卵巢癌细胞c13*转移侵袭的影响及机制  

Effect of BRMS1 on Migration,Invasion and Adhesion in Ovarian Cancer Cells(c13*)and Mechanisms in vitro

作　　者：饶玉梅[1] 方勇[1] 韩志强[1] 卢运萍[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,武汉430030

出　　处：《华中科技大学学报（医学版）》2011年第2期131-136,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家重点基础研究发展规划(973计划)(No.2009CB521808);国家自然科学基金(No.30700895)资助项目

摘　　要：目的探讨乳腺癌转移抑制基因1(BRMS1)对c13*细胞转移侵袭、粘附的影响及机制。方法由质粒PC-MV-HA-BRMS1中扩增目的基因BRMS1,将其插入pcDNA3.1(+)质粒中,构建pcDNA3.1(+)-BRMS1质粒。经酶切及测序鉴定正确后,转染c13*细胞,PCR、Real-time PCR、Western blot检测BRMS1的表达;划痕实验、Transwell实验、粘附实验检测c13*细胞运动、侵袭、粘附能力;Western blot检测整合素β1、p-AKT、AKT表达。结果成功构建了pcDNA3.1(+)-BRMS1质粒;转染目的基因的c13*细胞中,BRMS1的mRNA和蛋白表达均显著增加,细胞运动、侵袭、粘附能力降低,整合素β1及p-AKT表达均下降,AKT表达无改变。结论成功构建pcDNA3.1(+)-BRMS1质粒。BRMS1可能通过抑制整合素β1/AKT通路,抑制卵巢癌c13*细胞的运动、侵袭和粘附能力。Objective To construct and express the recombinant plasmid of pcDNA3.1（＋）-BRMS1 in ovarian cancer cells（c13＊）,and observe the effects of BRMS1 on migration,invasion and adhesion of c13＊ cells.Methods BRMS1 cDNA was obtained from the plasmid PCMV-HA-BRMS1 by PCR,and inserted into pcDNA3.1（＋）plasmid to generate a recombinant plasmid pcDNA3.1（＋）-BRMS1.The recombinant plasmid was verified by sequencing and enzyme digestion analysis,and transfected into c13＊ cells by using liposome.The BRMS1 expression was detected by PCR,real-time PCR,and Western blot after transfection.The effects of BRMS1 overexpression on the migration,invasion and adhesion of c13＊ were investigated by wound-healing,Transwell,and adhesion assay respectively.At last,the expression of integrin beta 1,p-AKT,and AKT was detected by Western blot after transfection.Results pcDNA3.1（＋）-BRMS1 recombinant plasmid was constructed and transfected into c13＊ cells successfully,and BRMS1 was over-expressed.The migration,invasion,and adhesion capability of c13＊ cells was decreased after pcDNA3.1（＋）-BRMS1 transfection.The expression of integrin beta 1 and p-AKT was down-regulated after transfection,but AKT expression had no significant change.Conclusion pcDNA3.1（＋）-BRMS1 recombinant plasmid was successfully constructed and transfected.The decreased migration,invasion and adhesion capability by enhancing the BRMS1 expression in c13＊ cells may be mediated through the altered expression of integrin beta 1/AKT signal pathway.

关 键 词：乳腺癌转移抑制基因1 整合素Β1 卵巢癌 肿瘤侵袭 肿瘤转移 

分 类 号：R737.31[医药卫生—肿瘤]