【摘要】 目的探讨5-氮-2’-脱氧胞苷(5-Aza-2’-deoxycytidine,5-Aza-CdR)对人卵巢癌细胞系SKOV3增殖、凋亡及错配修复基因HMLH1和HMSH2表达的影响。方法用0.5、5和50μmol/L特异性甲基转移酶抑制剂5-Aza-CdR处理SKOV3细胞3d,继续常规培养7d后,采用四唑盐(MTT)比色法检测生长活性,流式细胞术分析细胞的凋亡,半定量RT-PCR检测细胞HMLH1和HMSH2 mRNA的表达水平。结果人卵巢癌细胞系SKOV3经0.5、5和50μmol/L5-Aza-CdR处理后,均能明显抑制肿瘤细胞生长。各组细胞的凋亡率分别为10.59%±1.57%、17.52%±1.72%、34.10%±1.45%,显著高于对照组的5.35%±0.86%(P<0.01);且凋亡率与5-Aza-CdR剂量成正相关(F=227.6,P<0.01)。在SKOV3中HMLH1和HMSH2呈弱表达,经5-Aza-CdR处理后mRNA表达量有不同程度的增加,且与药物存在剂量依赖性。结论5-Aza-CdR可逆转卵巢癌细胞系中存在的HMLH1和HMSH2甲基化,DNA错配修复基因甲基化与卵巢癌的发生、发展有关。更多还原

【Abstract】 Objective To observe the effects of 5-Aza-2’-deoxycytidine(5-Aza-CdR)on cell proliferation and apoptosis in ovarian cancer cell line SKOV3 and the expression of mismatch repair gene HMLH1/HMSH2, and to investigate the potential mechanism of its antitumorigenesis. Methods Human ovarian cancer cell line SKOV3 was treated with 5-Aza-CdR(0.5, 5 and 50 μmol/L), a specific demethylation agent for 3 d, and then cultured in RPMI-1640 medium for 7 d.The growth of cells was examined by MTT assay. The apoptosis was analyzed by flow cytometry. The expression of HMLH1 and HMSH2 mRNA was observed by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). Results SKOV3 cells treated with 5-Aza-CdR displayed a slow growth rate in comparison with that of the control cells, The apoptosis rates of each group were 10.59%±1.57%、17.52%±1.72%、34.10%±1.45%,respectively,which were markedly higher than that of control(P<0.01).The apoptosis was also notably correlated with 5-Aza-CdR concentration(F=227.6,P<0.01). After 0.5, 5 and50 μmol/L 5-Aza-CdR treatment, the level of HMLH1 and HMSH2 mRNA expression was increased, and it was significantly]correlated with the concentration of 5-Aza-CdR. Conclusion 5-Aza-CdR can inhibit the proliferation and partly induce the apoptosis of human ovarian cancer cell line SKOV3 by inducing the expression of HMLH1 and HMSH2 genes,which may be inactivated by hypermethylation. Methylation of mismatch repair genes plays an important role in the carcinogenesis and progression of ovarian cancer.更多还原