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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 investigate the expression of VWCE gene in human breast cancer and its effect on the proliferation and migration of breast cancer cells in vitro. Methods: The expression of VWCE in 36 breast cancer tissues, adjacent normal tissues and 5 breast cancer cell lines was detected by real-time fluorescence quantitative PCR and Western blot respectively. The recombinant lentivirus overexpression of VWCE gene was constructed and transfected into MCF-7 and SK-BR-3 cells. The VWCE overexpression was verified by real-time fluorescence quantitative PCR and Western blot. The effects of VWCE overexpression on the proliferation and migration of breast cancer cells MCF-7 and SK-BR-3 were detected by CCK-8 and Transwell chamber assay. Results: The high expression rate of VWCE mRNA in breast cancer tissues was significantly lower than that in paracancer tissues (22% vs 78%, P <0.05), and significantly correlated with the staging of primary tumors (P <0.05). The VWCE protein The average expression level was also significantly lower than the adjacent tissue (P <0.05). The expression levels of VWCE mRNA and protein in 5 breast cancer cell lines were lower than that of immortalized mammary epithelial cell line (P <0.05). VWCE gene overexpression recombinant virus infected endogenous VWCE low expression of breast cancer MCF-7 and SK-BR-3 cells, the two kinds of cells were high expression of VWCE protein (P values were <0.05), cell proliferation and migration Ability were significantly reduced (P values were <0.05). Conclusion: VWCE is low expressed in breast cancer tissues and cells. VWCE overexpression can inhibit the proliferation and migration of breast cancer cells in vitro.