CAJ | 학술논문

目的：探究寡核苷酸MT01（ODN MT01）促进人骨髓间充质干细胞（hBMSCs）增殖的机制。方法将第三代hBMSCs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hBMSCs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。
목적：탐구과핵감산MT01（ODN MT01）촉진인골수간충질간세포（hBMSCs）증식적궤제。방법장제삼대hBMSCs이6.0×103/cm2접충우6공판，실험조화대조조각3개공，실험조가입ODN MT01사기종농도위2.0 mg/L，대조조가입등량PBS，련속검측3 d，응용MuseTM Cell Analyzer진행세포주기검측；병근거세포주기검측결과，분조동상，대상응처리후제1、2、3천적hBMSCs진행Cyclin A、Cyclin D1、CDK2화CDK4사충단백적실시형광정량검측。결과세포주기검측결과현시：여대조조상비，실험조처우G0/G1기적세포백분비강저，이처우S기화G2/M기적세포백분비승고，차이구유통계학의의（P＜0.01）；실시형광정량검측현시：여대조조상비，실험조중Cyclin A、Cyclin D1、CDK2화CDK4사충단백적표체유명현승고，차이구유통계학의의（P＜0.01）。결론여대조조상비ODN MT01강저료G0/G1기적세포백분비，제고료S기、G2/M기적세포백분비；기분자궤제가능단불국한우통과조고Cyclin A、Cyclin D1、CDK 2화CDK 4사충단백적표체이실현적。
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 mg/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.