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[目的]探讨中电导钙激活性钾离子通道(KCa3.1)在子宫内膜癌细胞迁移和侵袭中的作用。[方法]利用KCa3.1特异性阻断剂TRAM-34,通过划痕实验、趋向运动实验和Tran-swell小室侵袭实验了解KCa3.1在子宫内膜癌细胞株HEC-1-A、Ishikawa迁移和侵袭方面的作用,并用RT-PCR及Westernblot法检测TRAM-34作用前后MMP-2基因和蛋白表达水平的改变。[结果](1)TRAM-34可以显著性抑制HEC-1-A和Ishikawa细胞的迁移和侵袭,作用48h后HEC-1-A细胞和Ishikawa细胞加药组和对照组之间的划痕间距差异有统计学意义(t=28.23,P=0.00;t=35.29,P=0.00);趋向运动实验和体外侵袭实验均显示,两种细胞药物处理组和对照组的穿透滤膜细胞数目差异具有统计学意义(P均<0.05);(2)在HEC-1-A细胞株,药物处理组的MMP-2基因和蛋白表达水平随TRAM-34浓度升高而降低,与对照组相比差异均具有统计学意义(P均<0.05);在Ishikawa细胞,5μMTRAM-34处理组的MMP-2基因和蛋白表达水平与对照组相比无统计学差异,其他药物浓度处理组MMP-2基因和蛋白表达水平随TRAM-34浓度的升高而降低,与对照组相比差异有统计学意义(P均<0.05)。[结论]抑制KCa3.1活性可以降低子宫内膜癌细胞的迁移和侵袭能力,可能与下调MMP-2的表达有关。提示KCa3.1可能促进子宫内膜癌细胞的迁移和侵袭,它的表达和/或功能异常可能在子宫内膜癌的转移中起重要作用。
[Objective] To investigate the role of medium-conductance calcium-activated potassium channel (KCa3.1) in the migration and invasion of endometrial carcinoma cells. [Method] The KCa3.1 specific antagonist TRAM-34 was used to understand the migration of KCa3.1 in HEC-1-A and Ishikawa cells by using scratch test, tending movement test and Tran-swell cell invasion assay And invasion. RT-PCR and Western blotting were used to detect the expression of MMP-2 gene and protein before and after TRAM-34 treatment. [Results] (1) TRAM-34 markedly inhibited the migration and invasion of HEC-1-A and Ishikawa cells. After 48 h, the scratch spacing between HEC-1-A cells and Ishikawa cells in the drug- The difference was statistically significant (t = 28.23, P = 0.00; t = 35.29, P = 0.00). The trend movement test and in vitro invasion assay showed that the numbers of penetrating filter cells in the two cell drug treatment groups and the control group were different (P <0.05). (2) In HEC-1-A cell line, the expression of MMP-2 gene and protein in drug-treated group decreased with the increase of TRAM-34 concentration, compared with the control group (P <0.05). In Ishikawa cells, the expression of MMP-2 mRNA and protein in 5μM TRAM-34 group was not significantly different from that in the control group. The expression of MMP-2 gene in other drug concentration groups And protein expression decreased with the increase of TRAM-34 concentration, compared with the control group, the difference was statistically significant (P all <0.05). [Conclusion] Inhibition of KCa3.1 activity can reduce the migration and invasion of endometrial cancer cells, which may be related to the down-regulation of MMP-2 expression. It is suggested that KCa3.1 may promote the migration and invasion of endometrial cancer cells, and its expression and / or dysfunction may play an important role in the metastasis of endometrial cancer.