5-氮杂-2＇-脱氧胞苷对人卵巢癌细胞株RASSF1A基因表达的影响

Effects of 5-aza-2＇-deoxycitydine on expression of RASSF1A gene in human ovarian cancer cell line

目的 观察5-氮杂-2＇-脱氧胞苷（5-Aza-CdR）对体外培养的人卵巢癌细胞株（SKOV3、3AO） RASSF1A mRNA及蛋白表达的影响.方法 卵巢癌细胞株（SKOV3、3AO）经不同浓度（0.5、5、50 μmol/L）的5-Aza-CdR处理,未经处理的作为对照组,反转录聚合酶链反应（RT-PCR）及免疫印记（Western Blot）检测RASSF1A在mRNA和蛋白质水平表达的变化.结果 对照组细胞株SKOV3、3AO均可见RASSF1A mRNA及蛋白的表达,但表达较弱,经药物处理后的细胞RASSF1A mRNA及蛋白的表达较前明显增强.经不同浓度5-Aza-cdR处理的SKOV3细胞株中,随浓度增加,RASSF1A mRNA及蛋白的表达依次递增.经不同浓度5-Aza-cdR处理的3AO细胞株中,经0.5μmol/L 5-Aza-cdR处理后,RASSF1A mRNA的表达明显低于经5和50 μmol/L 5-Aza-cdR处理后的表达（t=-8.866,P=0.01;t=-12.256,P=0.000）;但经5、50 μmol/L 5-Aza-cdR分别处理的3AO细胞株RASSF1A mRNA的表达差异无统计学意义（t=0.431,P=0.689）.在3AO细胞株中,0.5 μmol/L组与5μmol/L组RASSF1A蛋白表达差异无统计学意义（t=-1.586,P=0.188）.结论 经过5-Aza-CdR处理后,卵巢癌细胞株（SKOV3、3AO） RASSF1A mRNA及蛋白的表达均明显增强,细胞增殖能力受到明显抑制,从而起到抑制肿瘤细胞生长,甚至促进肿瘤细胞凋亡的作用.

Objective To investigate the effect of 5-aza-2＇deoxycytidine （5-Aza-CdR） on proliferation and expression of RASSF1 A gene in human ovarian cancer cell line SKOV3 and 3AO.Methods SKOV3 and 3AO cells were treated with different concentrations （0.5,5,50 μmol/L） of DNA methyltransferase inhibitor 5-Aza-CdR.RT-PCR and Western Blot were adopted to detect expression of mRNA and protein of RASSF1A gene before and after treatment with 5-Aza-CdR respectively.Results Compared with control group,when the 5-Aza-CdR concentration was 0.5,5,50 μmol/L after drug treatment,human ovarian cancer cells could significantly inhibit tumor cell growth; SKOV3 and 3AO cells in control group were observed weaker expression of RASSF1A mRNA.After treated with 5-Aza-CdR,the expressions of RASSF1A mRNA were observed increased with the increase of the drug concentration.After treated with different concentration of 5-Aza-CdR,the expressions of RASSF1A mRNA treated with 0.5 μmol/L 5-Aza-cdR was lower than those treated with 5 and 50 μmol/L 5-Aza-cdR （t =-8.866,P =0.01 ; t =-12.256,P =0.000）.However,expressions of RASSF1A mRNA treated with 5 and 50 μmol/L 5-Aza-cdR respectively showed no statistical significance （t =0.431,P =0.689）.Expressions of RASSF1A protein treated with 0.5 μmol/L 5-Aza-cdR and 5 μmol/L 5-Aza-cdR didn＇t show statistically significant （t =-1.586,P =0.188）.Conclusion Expressions of RASSF1A mRNA and protein in SKOV3 and 3AO cells were evidently enhanced.As one kind of methyltransferases inhibitors,5-Aza-CdR can inhibit ovarian cancer cell line SKOV3,3AO growth through the RASSF1A promoter methylation,and thus promote their apoptosis.