摘要目的 研究雌激素调控紧密连接蛋白claudin-6表达和对乳腺癌细胞MCF-7细胞生物学行为的影响.方法 用17β-雌二醇(17β-E2)作用MCF-7细胞后,采用RT-PCR和免疫细胞化学法分析claudin-6表达与17β-E2的浓度和时间效应关系;采用Cell Counting Kit-8(CCK-8)检测17β-E2对MCF-7细胞增殖的影响;采用细胞划痕法检测17β-E2对MCF-7细胞迁移能力的影响.结果 RT-PCR结果显示,17β-E2能够诱导MCF-7细胞表达claudin-6,其诱导表达具有浓度和时间依赖性,其最佳浓度和时间分别为5 nmol/L和24 h;细胞免疫荧光法检测显示,17β-E2组claudin-6主要表达于细胞膜;CCK-8结果显示,5 nmol/L 17β-E2作用24 h可明显抑制MCF-7细胞的生长(P<0.05);细胞划痕实验显示,5 nmol/L 17β-E2作用24 h具有抑制MCF-7细胞迁移的趋势,但差异无统计学意义.结论 17β-E2可以调控claudin-6表达,并具有抑制MCF-7细胞的增殖和迁移的作用.推测17β-E2调控claudin-6表达可能在影响MCF-7细胞生物学行为的过程中发挥一定作用.

abstractsObjective To explore the role of estrogen in the regulation of the expression of claudin-6 and biological behavior in MCF-7 cells. Methods RT-PCR and immunocytochemistry were conducted to analyze the expression and localization of claudin-6 in MCF-7 cells treated with 17β-estradiol. CCK-8 kit assay and Scratch Test were conducted to analyze the capability of proliferation and migration of 17β-estradiol treated MCF-7 cells. Results RT-PCR analysis and immunocytochemistry showed that 17β-estradiol induced a concentration-and time-dependent effect on claudin-6. At 5 nmol/L and at 24 h, 17β-estradiol treatment led to an increased level of claudin-6, which was located in the membrane of MCF-7 cells. CCK-8 analysis showed a significant decrease in the capability of proliferation of MCF-7 cells compared with the control group ( P < 0. 05 ). Cells Scratch Test showed decreased migration capability of MCF-7 cells compared with the control group (P > 0. 05). Conclusions 17β-E2 might regulate the expression of claudin-6 and inhibit the proliferation and migration of MCF-7 cells. The inhibitory effects of 17β-E2 on growth and migration of MCF-7 cells may be mediated by claudin-6 expression regulation.