RNA干扰Semaphorin 4D表达对乳腺癌细胞增殖、周期及迁移的影响  

Effects of RNA inter ference targeting Semaphorin 4D on proliferation, cell cycle and migration of human breast cancer cells

作　　者：陈武桂 沈伟伟[1] 胡旭[1] 卓云云 毛德举[1] 初同伟[1] 

机构地区：[1]第三军医大学第二附属医院骨科,重庆400037

出　　处：《肿瘤》2015年第3期283-291,共9页Tumor

摘　　要：目的 :研究Semaphorin 4D(Sema 4D)表达沉默对乳腺癌细胞增殖、周期及迁移的影响,初步探讨该蛋白在乳腺癌发生和发展中的作用。方法 :实时荧光定量PCR和蛋白质印迹法检测Sema 4D在不同转移潜能的乳腺癌细胞株MDA-MB-231和MCF-7中的表达水平。采用短发夹RNA(short hairpin RNA,sh RNA)慢病毒感染法,建立Sema 4D稳定低表达的MDA-MB-231细胞系,然后采用CCK-8法、FCM法、划痕愈合实验及Transwell迁移实验检测细胞增殖、周期分布及迁移能力的变化。结果 :Sema 4D在MDA-MB-231细胞中表达水平明显高于MCF-7细胞(P<0.05);sh RNA慢病毒感染可明显下调MDA-MB-231细胞中Sema4D的表达;与空白对照组比较,Sema 4D干扰组的MDA-MB-231细胞增殖明显被抑制(P<0.05),G0/G1期细胞所占比例明显增高(P<0.05),而细胞迁移能力明显降低(P<0.05)。结论:Sema 4D在高转移潜能的乳腺癌MDA-MB-231细胞中高表达,sh RNA干扰Sema 4D表达可抑制该细胞的增殖和迁移,并阻滞细胞周期于G1期。Objective： To study the effects of Semaphorin 4D （Sema 4D） gene silencing on proliferation, cell cycle and migration of breast cancer cells, and investigate the role of Sema 4D protein in occurrence and development of breast cancer. Methods： The expression levels of Sema 4D mRNA and protein in two breast cancer cell lines MDA-MB-231 and MCF-7 with different metastatic potentials were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The MDA-MB-231 cell line with stable down-regulated expression of Sema 4D was established by infection of lentivirus containing Sema 4D-short hairpin RNA （shRNA）. Then the changes of cell proliferation, cell cycle distribution and migration ability were detected by cell counting kit-8 （CCK-8）, flow cytometry, wound healing assay and Transwell migration experiment, respectively. Results： The expression level of Sema 4D in MDA-MB-231 cells was significantly higher than that in MCF-7 cell （P 〈 0.05）. The expression of Sema 4D was significantly down-regulated in MDA-MB-231 cells after Sema 4D-shRNA lentivirus infection. Compared with the blank control group, the proliferation of MDA-MB-231 cells was significantly inhibited in Sema 4D-shRNA interference group （P 〈 0.05）, and the proportion of cells at G0/G1 phase was significantly increased （P 〈 0.05）, as well as the migration ability was obviously reduced （P 〈 0.05）. Conclusion： Sema 4D is expressed at a high level in breast cancer MDA-MB-231 cells with high metastatic potential. Down-regulation of Sema 4D expression by shRNA interference can significantly inhibit the proliferation and migration of MDA-MB-231 cells, and block the cell cycle at G1 phase.

关 键 词：乳腺肿瘤 信号素4D 细胞增殖 细胞周期 细胞迁移分析 

分 类 号：R737.9[医药卫生—肿瘤]