Promotion of miR-200b promoter methylation by MMC induces fibroblast apoptosis / 实用医学杂志

ABSTRACT

Objective To explore the mechanism of the role of mitomycin C(MMC)in regulating miR-200b expression and inducing fibroblasts apoptosis. Methods Fibroblasts cultured in vitro were treated with different concentrations of MMC for 5 min and continue culture for 24 h. The expression of miR-200b were analyzed by Real-time PCR. Cell apoptosis were observed using TUNEL staining. The expression of cleaved-PARP,Bax and Bcl-2 were detected by Western blot. The methylation level of miR-200b promoter were measured by BSP. Results After treated with MMC,The expression of miR-200b significantly downregulated.TUNEL Staining analysis demonstrated MMC could significantly induce human fibroblasts apoptosis. Western blot results showed cleaved-PARP,Bax increased and Bcl-2 decreased.The methylation ratio of miR-200b promotor increased and has a significant dose dependent. Conclusion MMC induced human fibroblasts apoptosis by promoting miR-200b promoter methylation.