摘要
Abstract
Objective To explore the possible mechanism and treatment countermeasure of cervical cancer by detecting promoter hypermethylation level of RASSF2A in four cervical cancer cells (HeLa,CaSki,SiHa and C33A) and analyzing the changes of mRNA and protein level of RASSF2A,methylation status and biological activity before and after demethylation. Meth-ods Real-time quantitative-polymerase chain reaction(RQ-PCR)was used to examine the expression of RASSF2A in cervical cancer cells before and after 5-aza-dC treatment. The promoter methylation of RASSF2A in the four cervical cells was detected by methylation specific PCR(MSP) methods. The expression changes of RASSF2A was detected by immunohistochemistry,and its influence on biological activity was observed with MTT and flow cytometry. Results The expression of HeLa and SiHa in cervical cancer lines were complete methylation,and the CaSki and C33A were partial methylation. The 5-aza-dC treatment could make part cervical cancer cells demethylation,as well as increase both the protein expression level and mRNA level of RASSF2A in cervical cancer cells. Biological behavior detection showed that the proliferation capacity and apoptosis rate of HeLa cell of cervi-cal cancer decreased under the induction of 5-aza-dC. Conclusion The methylation is one of great reasons for expression loss or decline of RASSF2A in cervical cancer cells. The 5-aza-dC can reverse RASSF2A gene promoter abnormal methylation level in cervical cancer cells,improve the expression level of mRNA and protein of RASSF2A gene,which show that the methylation level of RASSF2A,as a molecular marker,can evaluate whether the demethylation intervention is effective,and provide a new molecu-lar target for clinical treatment of cervical cancer.关键词
宫颈肿瘤/基因/DNA甲基化/5-aza-dC/细胞生物活性Key words
Uterine cervial neoplams/Gene/DNA methylation/5-aza-dC/Cell biological activity
