骨髓间充质干细胞对293T衰老细胞的作用机制研究

Study on mechanism of BMSCs on aging 293T cells

目的建立293T细胞衰老模型,评价脐带间充质干细胞对293T衰老细胞端粒及端粒酶相关基因TCAB1的影响。方法通过叔丁基过氧化氢(t-BHP)诱导获得293T细胞衰老模型,用猕猴骨髓间充质干细胞(BMSC)分别与293T细胞、293T细胞衰老模型共培养5d后,采用RT-qPCR法检测细胞端粒相对长度及端粒酶相关基因TCAB1的水平变化。结果通过200μmol/Lt-BHP处理293T细胞2h后,大量细胞出现衰老改变,而凋亡较少。与BMSC共培养5d后,293T细胞及衰老模型的端粒延长和TCAB1基因表达水平均有所升高(P<0.05)。结论t-BHP能够诱导293T细胞衰老,BMSC与293T细胞共培养具有提高细胞端粒酶活性和延长端粒的作用。

Objective To evaluate the effects of bone marrow mesenchymal stem cells(BMSCs)on telomeres and Telomerase Cajal body protein 1(TCAB1,gene related to telomerase)in aging 293T cells by establishing the aging model of 293T cells.Methods The aging model of 293T cells was induced with Tert-butyl Hydroperoxide(t-BHP),and the BMSCs of macaques were co-cultured with 293T cells and the aging model of 293T cells for five days.RT-qPCR method was used to detect the changes of relative telomere length and TCAB1 level.Results A large number of 293T cells showed aging changes with less apoptosis after being treated with 200μmol/L t-BHP for two hours.After co-culture with BMSCs for five days,the telomere elongation and gene expression levels of TCAB1 were improved in 293T cells and aging model of 293T cells(P<0.05).Conclusion Aging of 293T cells can be induced by t-BHP,and co-culture of BMSCs and 293T cells can improve the cell telomerase activity and lengthen telomeres.