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目的:探讨miR-203对人舌鳞癌细胞系Tca8113增殖能力的影响。方法:通过慢病毒转染技术构建miR-203过表达的舌鳞癌细胞模型。RT-PCR法检测miR-203过表达慢病毒转染前后miR-203的表达水平;MTT方法检测miR-203过表达(micro up组)及阴性对照(NC组)2组细胞的增殖能力,连续检测5d,酶标仪检测A490值,绘制细胞增殖曲线;PI-FACS细胞周期检测miR-203过表达后对DNA合成和细胞周期的影响,取处于对数生长期的细胞,70%乙醇固定>1h,细胞染色,流式细胞仪检测。结果:成功建立了miR-203过表达舌鳞癌细胞模型。定量PCR结果显示,Tca8113细胞中,micro up组miR-203表达丰度是NC组的286.262倍,t=-34.879,P=0.001。MTT检测结果表明,与NC组相比,micro up组细胞增殖能力明显降低;检测第4天micro up组A490值为0.492±0.011,NC组为0.681±0.009,t=-23.463,P<0.001。PI-FACS细胞周期检测结果显示,micro up组DNA合成前期(G1期)细胞所占百分率为(58.98±0.56)%,较NC组的(53.86±0.96)%明显增高,t=-7.989,P=0.001;而DNA合成期(S期)、合成后期(G2期)及有丝分裂期(M期)细胞所占百分率降低。结论:miR-203可能通过将细胞周期阻滞于G1期,从而抑制舌鳞癌细胞系Tca8113的细胞增殖能力。
Objective: To investigate the effect of miR-203 on the proliferation of human tongue squamous cell carcinoma cell line Tca8113. Methods: The miR-203 overexpression tongue squamous cell carcinoma cell model was constructed by lentiviral transfection technique. The expression of miR-203 was detected by RT-PCR before and after transfection with miR-203 overexpression lentivirus. The proliferative ability of miR-203 over-expression (micro up group) and negative control group (NC group) After 5 days of detection, the A490 value was detected by microplate reader and the cell proliferation curve was drawn. PI-FACS cell cycle was used to detect the effect of miR-203 over-expression on DNA synthesis and cell cycle. Cells in logarithmic growth phase, 70% ethanol fixation> 1h, cell staining, flow cytometry. Results: miR-203 overexpression tongue squamous cell carcinoma cell model was established successfully. Quantitative PCR results showed that the expression level of miR-203 in micro-up group in Tca8113 cells was 286.262 times of that in NC group, t = -34.879, P = 0.001. The results of MTT assay showed that compared with NC group, the proliferation ability of micro up group was significantly decreased. The A490 value of micro up group was 0.492 ± 0.011 on day 4, 0.681 ± 0.009 for NC group, t = -23.463, P <0.001. The PI-FACS cell cycle assay showed that the percentage of cells in the micro up group (58.98 ± 0.56%) was significantly higher than that in the NC group (53.86 ± 0.96)%, t = -7.989, P = 0.001; while the percentage of cells in DNA synthesis phase (S phase), synthesis phase (G2 phase) and mitosis phase (M phase) decreased. Conclusion: miR-203 may inhibit cell proliferation of tongue squamous cell carcinoma cell line Tca8113 by blocking the cell cycle in G1 phase.