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目的:本实验检测PRDM5在HTB-182、A549、HBE正常支气管上皮细胞系中甲基化状态及去甲基化处理对其表达的影响。方法:运用MSP甲基化特异性PCR检测PRDM5在A549、HTB-182和HBE正常支气管上皮细胞系的甲基化状态,再加入不同浓度的去甲基化药物检测PRDM5在A549、HTB-182和HBE正常支气管上皮细胞的甲基化状态,用RT-PCR检测PRDM5在加药前后的mRNA表达水平;用Western-Blot检测PRDM5在加药前后的蛋白表达水平。结果:在肺肿瘤细胞系中,PRDM5存在不同程度的高甲基化,加入不同浓度去甲基化药物后,甲基化表达水平逐渐减弱(P<0.05),mRNA表达水平逐渐增强(P<0.05),蛋白表达水平也逐渐增强(P<0.05)。结论:在肺癌细胞系中PRDM5启动子高甲基化是导致PRDM5表达水平降低的主要原因,PRD-M5启动子去甲基化可能成为肺癌治疗的新靶点。
Objective: To investigate the effect of methylation status and demethylation of PRDM5 in HTB-182, A549 and HBE normal bronchial epithelial cell lines. Methods: The methylation status of PRDM5 in A549, HTB-182 and HBE normal bronchial epithelial cell lines was detected by MSP methylation-specific PCR. Different concentrations of demethylation drugs were added to detect the expression of PRDM5 in A549, HTB-182 and HBE normal bronchial epithelial cells methylation status detected by RT-PCR PRDM5 before and after dosing mRNA expression levels; using Western-Blot PRDM5 dosing before and after the protein expression levels. Results: The expression of PRDM5 in different degrees of hypermethylation was significantly different in lung cancer cell lines. The methylation level of PRDM5 decreased gradually (P <0.05) and the mRNA level of PRDM5 gradually increased (P <0.05) , Protein expression level also gradually increased (P <0.05). CONCLUSION: PRDM5 promoter hypermethylation in lung cancer cell lines is the main reason leading to the decrease of PRDM5 expression. Demethylation of PRD-M5 promoter may be a new target for the treatment of lung cancer.