5-氮胞苷对宫颈癌细胞系DAPK1异常甲基化的影响

目的　观察宫颈癌细胞系SiHa及HeLa中凋亡相关蛋白激酶 1(DAPK1)的表达与甲基化修饰之间的相关性,以及5 氮胞苷对细胞DAPK1表达和生长的影响。方法　应用甲基化特异性PCR法分析细胞中DAPK1的甲基化状态;采用RT- PCR及免疫组化SABC法检测不同浓度 5- 氮胞苷干预对宫颈癌细胞 DAPK1 mRNA及蛋白表达的影响;MTT法观察5 -氮胞苷对细胞增殖的影响。结果　DAPK1基因在宫颈癌细胞 SiHa中有甲基化修饰,5- 氮胞苷干预能使其甲基化的DAPK1基因重新表达并抑制细胞生长。结论　DAPK1 的甲基化修饰是宫颈癌发生重要机制之一,5 -氮胞苷能使甲基化的DAPK1去除甲基化修饰,重新表达并恢复抑癌作用。

Effect of 5-azacytidine on the methylation of DAPK1 in cervical carcinoma cell line

Objective To investigate the correlation between methylation status and death-associated protein kinase 1 (DAPK1) in SiHa and Hela cell line of cervical carcinoma and intervention of DNA methyltransferase inhibitor 5-azacytidine on the expression of DAPK1 and the proliferation of the cells. Methods DAPK1 methylation status was analyzed using methylation-specific PCR methods. The expression of mRNA and protein of DAPK1 were analyzed by RT-PCR and SABC methods after the treatment with 5-azacytidine. MTT assay was used to observe the changes of proliferation activity of the cells after 5-azacytidine treatment. Results DAPK1 genes were methylated and did not express in SiHa cells in the cervical carcinoma. Its expression could be restored by 5-azacytidine. MTT assay showed 5-azacytidine could weaken the proliferation of cancerous cells. Conclusion DAPK1 methylation plays an important role in the carcinogenesis of cervical cells and can reexpress after the treatment with 5-azacytidine which also restored its inhibitory function on carcinoma.