摘要
目的探讨BRG1基因对乳腺癌细胞迁移及侵袭的影响及机制。方法BRG1小干扰RNA（siRNA）、对照siRNA分别转染人乳腺癌MDA—MB-231和BT-549细胞，细胞迁移实验观察迁移能力；细胞侵袭实验观察侵袭能力；Westernblot检测基质金属蛋白酶（MMPs）及组织金属蛋白酶抑制剂（TIMPs）蛋白表达；明胶酶谱实验检测细胞分泌MMPs能力。结果与对照组比较，BRG1siRNA处理组MDA-MB-231和BT-549两种细胞的迁移能力分别减少了58％；侵袭能力分别减少了66％和42％；Westernblot检测结果显示，BRG1．siRNA组两种乳腺癌细胞的MMP-2表达水平分别下降了24％和28％，MMP-9分别下降了30％和46％。TIMP2表达水平分别升高了0．72倍和0．55倍，TIMP1表达水平分别升高了1．75倍和3．09倍。明胶酶谱法检测结果显示，BRG1-siRNA组两种乳腺癌细胞中MMP-2活性分别下降了17％和20％，MMP-9活性分别下降了21％和11％。所有比较差异均有统计学意义（P〈0．05）。结论BRG1基因干涉后通过上调TIMP1、TIMP2表达进而抑制MMP-2、MMP-9表达及分泌，最终抑制乳腺癌细胞的侵袭能力。
Objective To investigate the role of brahma-related gene 1 （ BRG1 ） on human breast cancer cell migration and invasion and its molecular mechanisms. Methods The BRG1 small interfering RNA （siRNA） were transferred into MDA-MB-231 and BT-549 breast cancer cells. The migration abilities of two breast cancer cells were detected by cell migration assay. The invasive abilities were detected by cell invasion assay. The matrix metalloproteinase （MMP） and tissue inhibitorof metalloproteinase （TIMPs） proteins expressions were detected by western blotting and the MMPs activities were detected by gelatin zy- mography. Results Compared to the control siRNA treatment, BRG1 interference could drastically de- creased cell migration abilities of both cells by 58%, and the cell invasive abilities of both cells by 66% and 42%, respectively. The expression levels of MMP-2 protein were drastically decreased by 24% and 28% and the expression levels of MMP-9 protein were drastically decreased by 30% and 46% in the two breast cancer cells after BRG1 interference, but TIMP-1 and TIMP-2 protein expression level was signifi- candy increased. The MMP-2 activity was significantly suppressed in the two breast cancer cells by 17% and 20% , and the MMP-9 activity was significantly suppressed by 21% and 11% , respectively, compared to the control cells. Conclusion Silence of BRG1 could suppress human breast cancer cell invasion through up-regulation of TIMP-1, 2 and down-regulation of MMP-2, 9.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第6期1210-1212,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（81201636）
江苏省自然科学基金资助项目（BK2012139）
关键词
BRG1基因
乳腺癌
侵袭
基质金属蛋白酶
组织金属蛋白酶抑制剂
Brahma-related gene 1
Breast cancer
Invasion
Matrix metalloproteinase
Tissue inhibitorof metalloproteinase
