17β-雌二醇对紧密连接蛋白claudin-6表达及乳腺癌MCF-7细胞生物学行为的影响

Effects of 17β-estradiol on proliferation and migration of MCF-7 cell by regulating expression of claudin-6

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（17p—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17p—E2的浓度和时间效应关系；采用CellCountingKit-8（CCK-8）检测17B—E2对MCF-7细胞增殖的影响；采用细胞划痕法检测17B—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5nmol／L和24h；细胞免疫荧光法检测显示，17β—E2组claudin45主要表达于细胞膜；CCK-8结果显示，5mnol／L17β—E2作用24h可明显抑制MCF-7细胞的生长（P〈0．05）；细胞划痕实验显示，5nmol／L17β—E2作用24h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。

Objective To exptore the rote 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 1715-estradiol induced a concentration-and time-dependent effect on claudin-6. At 5 nmol, 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.