Effect of DNA methylation inhibitor on RASSF1A genes expression in non-small cell lung cancer cell l

来源 :第三届东北三省暨内蒙古自治区免疫学学术会议 | 被引量 : 0次 | 上传用户:shgandang
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  To compare the tumor suppressor gene RASSF1As methylation status and its mRNA expression level between non-small lung cancer cell line A549 and anti-Cisplatin cell strain A549DDP by Methylation Specific Polymerase Chain Reaction (MSP) and Reverse Transcription Polymerase Chain Reaction (RT-PCR), respectively.To reverse methylation of A549DDP by series of concentration of 5-Aza-2-deoxycytidine (5-Aza-cdR), that is DNA methyltransferase inhibitor.And then, Cell proliferation was detected by MTT assay.Cell apoptosis and the changes of cell cycle were detected by flow cytometry.The methylation status of RASSF1A gene before and after drug intervertion and the expression of mRNA were detected by MSP method and RT-PCR.Through these experiments, we come to some conclusions: there are obvious differences between A549 and A549DDP in RASSF1As methylation status and its mRNAs expression.48 hrs intervention of5-Aza-CdR showed a remarkable inhibition action towards A549DDP.And then, cell apoptosis rate was increased with the increase of drug dosage in a dose dependent manner.At the same time, the S stage of A549DDP cell declined in proportion with G2/M stage, increased in proportion with G0/G1stage, and cell motility was blocked in G0/G1 stage.All of A549DDP cells showed unmethylated expression, its high methylation status was reversed in a dose-dependent manner within a certain range.
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