人乳腺癌中VWCE基因的表达及其功能研究  

Expression and function of VWCE gene in human breast cancer

作　　者：万丽丽[1,2] 刘洋[2] 刘丽桃 朱婷[2] 饶茜[2] 周光纪 谢妮[2] 

机构地区：[1]广东医科大学基础医学院,广东东莞523000 [2]深圳市第二人民医院中心实验室,广东深圳518035

出　　处：《肿瘤》2017年第7期742-749,共8页Tumor

基　　金：深圳市国际合作研究项目(编号:GJHZ20160301163138685)~~

摘　　要：目的 :探讨人乳腺癌组织中VWCE(von Willebrand factor C and epidermal growth factor domains)基因的表达情况,以及其过表达对乳腺癌细胞体外增殖和迁移能力的影响。方法:采用实时荧光定量PCR和蛋白质印迹法分别检测36例乳腺癌组织及其癌旁正常组织和5种乳腺癌细胞系中VWCE的表达情况。构建VWCE基因过表达的重组慢病毒,将其感染乳腺癌MCF-7和SK-BR-3细胞后,采用实时荧光定量PCR和蛋白质印迹法验证VWCE过表达。分别采用CCK-8和Transwell小室法检测VWCE过表达对乳腺癌MCF-7和SK-BR-3细胞增殖和迁移能力的影响。结果:乳腺癌组织中VWCE mRNA的高表达率明显低于癌旁组织(22%vs78%,P<0.05),并与原发肿瘤分期明显相关(P<0.05);乳腺癌组织中VWCE蛋白的平均表达水平也明显低于癌旁组织(P<0.05)。5种乳腺癌细胞系中VWCE mRNA和蛋白的表达水平均低于永生化乳腺上皮细胞系(P值均<0.05)。将VWCE基因过表达重组病毒感染内源性VWCE低表达的乳腺癌MCF-7和SK-BR-3细胞后,2种细胞中VWCE蛋白均高表达(P值均<0.05),细胞增殖和迁移能力均明显降低(P值均<0.05)。结论:VWCE在乳腺癌组织和细胞中均低表达。VWCE过表达可以抑制乳腺癌细胞的体外增殖和迁移。Objective： To determine the expression of von Willebrand factor C and epidermal growth factor domains （VWCE） gene in human breast cancer tissues,and to assess the effects of VWCE overexpression on proliferation and migration of breast cancer cells in vitro. Methods： The expression levels of VWCE mRNA and protein in 36 pairs of human breast cancer tissues and their adjacent normal tissues as well as 5 kinds of breast cancer cell lines were detected by real-time fluorescent quantitative PCR and Western blotting,respectively.The recombinant lentivirus carrying VWCE gene was constructed and infected into breast cancer MCF-7 and SK-BR-3 cells.After the overexpression of VWCE gene was verified by real-time fluorescent quantitative PCR and Western blotting,the proliferation and migration of breast cancer cells were detected by CCK-8 and Transwell chamber assay,respectively. Results： The high expression rate of VWCE mRNA in breast cancer tissues was significantly lower than that in the corresponding adjacent tissues （22%vs 78%,P 〈 0.05）,and was related with the primary tumor stage （P 〈 0.05）.The mean expression level of VWCE protein in breast cancer tissues was lower than that in the corresponding adjacent tissues （P 〈 0.05）.The expression levels of VWCE mRNA and protein in 5 kinds of breast cancer cell lines were lower than those in immortalized mammary epithelial cell line （all P 〈 0.05）,respectively.After the breast cancer MCF-7 and SK-BR-3 cells were infected with VWCE overexpression lentivirus,the expression of VWCE protein was significantly increased （both P 〈 0.05）,while the cell proliferation and migration were significantly inhibited （all P 〈 0.05）. Conclusion： VWCE is low expressed in human breast cancer tissues and cells.Overexpression of VWCE can inhibit the proliferation and migration of breast cancer cells in vitro.

关 键 词：乳腺肿瘤 VONWILLEBRAND因子 基因表达 细胞增殖 细胞迁移分析 vWCE 

分 类 号：R737.9[医药卫生—肿瘤]