5-Aza-CdR对人结肠癌Caco-2细胞系P16基因甲基化状态及其生物学表型的影响

目的 观察人结肠癌Caco-2细胞系P16基因启动子区甲基化状态,并探讨去甲基化制剂5-氮杂-2-脱氧胞苷(5-Aza-CdR)诱导高甲基化失活的P16基因重新表达的可能性及其对细胞生长的影响.方法 用不同浓度的5-Aza-CdR处理Caco-2细胞系,MSP法检测用药前后P16基因的甲基化状态,RT-PCR方法检测P...

Effects of 5-Aza-CdR Methylation State of P16 Gene and Biological Phenotype in Human Colorectal Cancer Cell Line Caco-2

Objective To investigate the P16 gene methylation state in human colorectal cancer cell line Caco-2 and explore the possibility of re-expression of the hypermethylated and silenced P16 gene. Methods cell line Caco-2 treated with different concent rations of DNA methyltransferase inhibitor 5-Aza-CdR, MSP was used to detect promoter methylation state of P16 gene and RT-PCR were used to detect re-expression of mRNA before and after treatment with 5-Aza-CdR respectively. Cell growth speed was measured using MTT assay, cell cycle distribution and apoptosis rate were estimated using ?ow cytometry. Results Promoter hypermethylation of the P16 gene was detected in human colorectal cancer cell line Caco-2, After treatment with 5-Aza-CdR, the promoter region of the P16 gene exhibited a demethylation state, and 5-Aza-CdR induces P16 gene re-expression. 5-Aza-CdR effectively reversed the hypermethylation status of CpG island,induced colorectal cancer cell apoptosis and cell cycle arrest were observed in a dose -dependent manner. Conclusion P16 gene can be reversed by demethylation agent 5-Aza-CdR. demethylation agent can regulate the expression of the P16 gene,and effectively inhibits cell growth.