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目的:观察甲基转移酶抑制剂5-氮-2’脱氧胞苷(5-Aza-CdR)对人卵巢癌细胞系OVCAR3和Anglne RIZ1基因启动子区甲基化状态及表达的影响,探讨人卵巢癌细胞系OVCAR3和AnglneRIZ1基因失活的机制及5-Aza-CdR对RIZ1基因表达的调控。方法:用5-Aza-CdR(浓度为5μmol/L)处理体外培养的人卵巢癌细胞系OVCAR3和Anglne后,采用MSP法检测用药前后细胞中RIZl基因的甲基化状态,RT-PCR及Western-blot法检测用药前后细胞中RIZ1基因mRNA及蛋白表达的变化。结果:在人卵巢癌细胞系OVCAR3和Anglne中,RIZ1基因启动子区呈异常甲基化状态,RIZ1基因mRNA及蛋白表达缺失,经过5-Aza-CdR处理后,RIZ1基因启动子区呈非甲基化状态,其mRNA及蛋白重表达。结论:5-Aza-CdR可改变人卵巢癌细胞系OVCAR3和Anglne中RIZ1基因甲基化状态,从而调控癌细胞中RIZ1基因的表达。
AIM: To observe the effect of 5-Aza-CdR on the methylation status and expression of OVCAR3 and Anglne RIZ1 promoter in human ovarian cancer cell lines The mechanism of inactivation of OVCAR3 and AnglneRIZ1 genes in ovarian cancer cell lines and the regulation of RIZ1 gene expression by 5-Aza-CdR. Methods: The ovarian cancer cell lines OVCAR3 and Anglne were treated with 5-Aza-CdR (5μmol / L), and the methylation status of RIZ1 gene was detected by MSP method. RT-PCR and Western blot -blot assay before and after treatment of cells RIZ1 gene mRNA and protein expression changes. Results: In ovarian cancer cell lines OVCAR3 and Anglne, the promoter region of RIZ1 gene was abnormally methylated and the mRNA and protein expression of RIZ1 gene were absent. After treatment with 5-Aza-CdR, the promoter region of RIZ1 gene was non-A The state of the state, its mRNA and protein re-expression. CONCLUSION: 5-Aza-CdR can regulate the methylation status of RIZ1 gene in human ovarian cancer cell lines OVCAR3 and Anglne and thus regulate the expression of RIZ1 gene in cancer cells.