5-氮-2′-脱氧胞苷对DU145前列腺癌细胞TIMP-3基因去甲基化作用及侵袭能力的影响

目的观察5-氮-2′-脱氧胞苷(5Aza-dc)对DU145前列腺癌细胞金属蛋白酶组织抑制因子3(TI MP-3)启动子去甲基化作用及侵袭能力的影响。方法用5Aza-dc处理DU145前列腺癌细胞,Transwell检测癌细胞的侵袭及运动能力,反转录-聚合酶链反应(RT-PCR)检测TI MP-3 mRNA的表达,Western印迹检测TI MP-3的蛋白表达。结果5Aza-dc作用后的DU145细胞侵袭及运动能力降低;TI MP-3 mRNA及蛋白恢复表达;TI MP-3启动子区去甲基化。结论5Aza-dc可以使DU145前列腺癌细胞TI MP-3启动子区去甲基化,使TI MP-3基因重新表达,恢复其抑制肿瘤侵袭和移动的能力。

Effect of 5-Aza-2′-deoxycytidine on demethylation of tissue inhibitor of metalloproteinase-3 gene promoter in prostate cancer line

Objective To observe the effect of 5Aza-dc on demethylation of TIMP-3 gene promoter in prostate cancer line.Methods Prostate cancer line DU145 with methylation of TIMP-3 promoter gene was treated with 5-Aza-2′-deoxycytidine (5Aza-dc).Invasion ability and motility of the cells was detected by Transwell experiments.Expressions of TIMP-3 protein and mRNA were detected by western blotting and RT-PCR, respectively.TIMP-3 gene promoter methylation was detected by methylation-specific PCR (MSP).Results After treatment with 5Aza-dc,Invasion ability and motility of DU145 cell line were declined;the expression of TIMP-3 protein and mRNA were increased in DU145 cell line;methylation of TIMP-3 promoter gene was not detectable in DU145 cell line.Conclusion 5Aza-dc induces demethylation in TIMP-3 promoter gene,restores TIMP-3 gene and protein expression,and inhibits invasion ability and motility of the prostate cancer line.