ODNMT01促进骨髓间充质干细胞增殖机制的研究

The molecular mechanism underlying the effect of ODN MT01 on h BMSC proliferation

目的探究寡核苷酸MT01(ODN MT01)促进人骨髓间充质干细胞(h BMSCs)增殖的机制。方法将第三代h BMSCs以6.0×103/cm2接种于6孔板,实验组和对照组各3个孔,实验组加入ODN MT01使其终浓度为2.0 mg/L,对照组加入等量PBS,连续检测3 d,应用MuseTM Cell Analyzer进行细胞周期检测;并根据细胞周期检测结果,分组同上,对相应处理后第1、2、3天的h BMSCs进行Cyclin A、Cyclin D1、CDK2和CDK4四种蛋白的实时荧光定量检测。结果细胞周期检测结果显示:与对照组相比,实验组处于G0/G1期的细胞百分比降低,而处于S期和G2/M期的细胞百分比升高,差异具有统计学意义(P<0.01);实时荧光定量检测显示:与对照组相比,实验组中Cyclin A、Cyclin D1、CDK2和CDK4四种蛋白的表达有明显升高,差异具有统计学意义(P<0.01)。结论与对照组相比ODN MT01降低了G0/G1期的细胞百分比,提高了S期、G2/M期的细胞百分比;其分子机制可能但不局限于通过调高Cyclin A、Cyclin D1、CDK 2和CDK 4四种蛋白的表达而实现的。

Objective The objectives of this study were to investigate the mechanism of action of the ODN MT01 that has the greatest effect on the hBMSCs. Methods hBMSCs were isolated, cultured to the third passage and were seeded at 6.0× 103/cm2 into 6-well plate. A total of 3 experimental groups and 3 control group of hBMSCs were established and treated with ODN types MT01, at concentrations of 2.0 rag/L; the control group was treated with an equal volume of PBS. Cell cycle analysis was done on days 1, 2 and 3 after ODN MT01 treatment; the expressions of Cyclin A, Cyclin D1, cyclin dependent kinase （CDK）2 and CDK4 in hBMSCs were measured on day 1, 2, 3 after treatment using fluorescent quantitative real-time PCR. Results The percentage of cells in phase G0/G1 was significantly reduced, and the percentage of cells in phases S and G2/M was significantly increased （P〈0.01） after treatment with 2.0 mg/L MT01 compared to the control group. Furthermore, the expressions of Cyclin A, Cyclin D1, CDK2 and CDK4 were significantly elevated （P〈0.01） compared with the control group, Conclusion A 2.0 mg/L concentration of MT01 significantly promotes hBMSCs proliferation as evidenced by the decrease in the percentage of cells in phase G0/G1 and the increase in the percentage of cells in phases S and G2/M. The underlying molecular mechanisms may include, but are not limited to, elevated expressions of Cyclin A, Cyclin D1, CDK2 and CDK4.