MRTF-A参与一氧化氮诱导的乳腺癌迁移相关基因表达  

MRTF-A Involved Expression of Nitric Oxide Induced Migration-related Gene in Breast Cancer Cells

作　　者：卢玉琳 胡娟 李智 何红鹏[1] LU Yulin;HU Juan;LI Zhi;HE Hongpeng(Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,Tianjin Key Laboratory of Industrial Microbiology,College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China)

机构地区：[1]工业发酵微生物教育部重点实验室,天津市工业微生物重点实验室,天津科技大学生物工程学院,天津300457

出　　处：《天津科技大学学报》2020年第5期8-14,共7页Journal of Tianjin University of Science & Technology

基　　金：天津市自然科学基金资助项目(18JCYBJC91500)。

摘　　要：为研究NO对乳腺癌细胞迁移能力的影响及其分子机制,本研究首先用梯度浓度NO供体SNP处理乳腺癌细胞MCF-7,MTT法检测细胞活性,发现高浓度SNP抑制细胞增殖;PI-Annexin V和Hoechst细胞核染色检测结果显示,高浓度NO促进细胞凋亡.用低浓度SNP处理细胞,划痕愈合及小室迁移实验结果显示,低浓度SNP促进细胞迁移;RT-qPCR和Western blot结果显示,低浓度SNP上调转录调控辅因子MRTF-A及其下游的迁移相关基因MYL9、MYH9和CYR61的表达.在MCF-7细胞敲低NOS2基因同样导致MRTF-A、MYL9、MYH9及CYR61表达下降;敲低MRTF-A则SNP不再促进上述迁移相关基因的表达.以上实验结果提示:NO的肿瘤生物学效应与NO浓度密切相关,细胞内源性或外源性低浓度NO可能通过刺激迁移相关基因表达而促进乳腺癌细胞转移,而MRTF-A参与了NO诱导的迁移相关基因表达.To study the effect of NO on the migration ability of breast cancer cells and its molecular mechanism,breast cancer cells MCF-7 were treated with serial diluted NO donor SNP.The results of MTT assay showed that,at higher concentrations,SNP inhibited the proliferation of MCF-7 cells and induced apoptosis as demonstrated by PI-Annexin and Hoechst nuclear staining assays.Wound-healing and Transwell chamber migration assays revealed that the low concentrations of NO promoted the migration of MCF-7 cells.The results of RT-qPCR and Western blot showed that,at low concentrations,NO stimulated the expression of a transcription co-activator MRTF-A and its downstream migration-related genes MYL9,MYH9 and CYR61.In MCF-7 cells,the expression of MRTF-A,MYL9,MYH9 and CYR61 also decreased following the knockdown of NO synthetase gene NOS2.Knockdown of MRTF-A in MCF-7 cells reversed the inducing effect of SNP on the expression of migration-related genes.These results suggest that the biological effects of NO are concentration-dependent.At low concentrations,both endogenous and exogenous NO promoted the migration of breast cancer cells by stimulating the expression of migration-related genes which were regulated by MRTF-A.

关 键 词：一氧化氮 乳腺癌 MRTF-A 细胞迁移 

分 类 号：R737.9[医药卫生—肿瘤]