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目的探讨去甲基化药物5-氮-2’-脱氧胞苷(5-aza-2’-deoxycytidine,DAC)对膀胱肿瘤细胞生长的抑制作用。方法采用WST-1法测定DAC对膀胱癌细胞株的生长影响作用,流式细胞仪检测其在诱导细胞凋亡与细胞周期捕获中的效果,APOPCYTO半胱天冬酶(Caspase)色度法分析DAC处理后Caspase 3及Caspase 9的活性,Western blot检测DAC处理后增殖细胞核抗原(PCNA)的表达情况。结果 DAC对膀胱肿瘤细胞的生长具有抑制作用,药物作用呈剂量依赖性,并且该作用不受p53表型影响。DAC不能诱导膀胱肿瘤细胞凋亡,细胞中Caspase 3、9的活性也未见激活,但DAC可以抑制膀胱癌细胞的增殖能力,PCNA蛋白的表达量呈剂量依存性降低,发生G2/M期捕获。用0、1和8μmol/LDAC处理RT112后G2/M期细胞分别为(36.3±3.4)%、(46.2±4.6)%和(56.5±6.2)%;处理TCCsup后为(37.5±3.8)%、(48.4±4.9)%和(60.1±6.7)%。结论 DAC对膀胱肿瘤细胞生长具有抑制作用,可能成为将来治疗膀胱恶性肿瘤的一种新型药物。
Objective To investigate the inhibitory effect of demethylation drug 5-aza-2’-deoxycytidine (DAC) on the growth of bladder tumor cells. Methods WST-1 method was used to determine the effects of DAC on the growth of bladder cancer cell lines. Flow cytometry was used to detect the effect of DAC on apoptosis induction and cell cycle arrest. APOPCYTO Caspase colorimetry Caspase 3 and Caspase 9 activity after DAC treatment, and the expression of proliferating cell nuclear antigen (PCNA) after DAC treatment by Western blot. Results DAC inhibited the growth of bladder tumor cells in a dose-dependent manner and the effect was not affected by p53 phenotype. DAC can not induce apoptosis of bladder tumor cells, but no activation of Caspase 3 and 9 in cells. However, DAC can inhibit the proliferation of bladder cancer cells, and the expression of PCNA protein is dose-dependently reduced and G2 / M phase arrest occurs . The percentages of cells in G2 / M phase were (36.3 ± 3.4)%, (46.2 ± 4.6)% and (56.5 ± 6.2)% respectively after treatment with RT112 at 0, 1 and 8μmol / L and 37.5 ± 3.8% (48.4 ± 4.9)% and (60.1 ± 6.7)%, respectively. Conclusion DAC can inhibit the growth of bladder tumor cells and may be a new drug for the treatment of malignant bladder tumors in the future.