miR-203对Tca8113增殖能力影响观察

发布时间：2018-07-25 16:00

【摘要】：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,P0.001。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。
[Abstract]:Objective: to investigate the effect of miR-203 on the proliferation of human tongue squamous cell carcinoma cell line Tca8113. Methods: Lentivirus transfection technique was used to construct tongue squamous carcinoma cell model with miR-203 overexpression. RT-PCR was used to detect the expression level of miR-203 before and after lentivirus transfection. MiR-203 overexpression (micro up group and negative control group (NC group) were used to detect the proliferative ability of the two groups. After 5 days of continuous detection, the A490 value was detected by enzyme labeling instrument. The cell cycle of PI-FACS was plotted to detect the effect of miR-203 overexpression on the synthesis of DNA and cell cycle. The cells in logarithmic growth phase were fixed with 70% ethanol for 1 hour. The cells were stained and detected by flow cytometry. Results: miR-203 overexpression of tongue squamous carcinoma cell model was successfully established. The results of quantitative PCR showed that the abundance of miR-203 in the micro up group was 286.262 times higher than that in the NC group. The results showed that the proliferation ability of the micro up group was significantly lower than that of the NC group. On the 4th day, the A490 value of micro up group was 0.492 卤0.011%, which was 0.681 卤0.009 t ~ (-23.463) (P 0.001) P0.001. Results showed that the percentage of DNA (G _ 1 phase) was (58.98 卤0.56) in the micro up group, which was significantly higher than that in the NC group (53.86 卤0.96)%, while the DNA synthesis phase (S phase) and the late synthetic phase (G _ 2) were significantly higher than those in the NC group (P < 0.05). The percentage of mitotic (M phase) cells decreased. Conclusion the cell proliferation of tongue squamous cell carcinoma cell line Tca8113 may be inhibited by blocking cell cycle at G1 phase.
【作者单位】：首都医科大学附属北京同仁医院耳鼻咽喉头颈外科耳鼻咽喉头颈科学教育部重点实验室;
【基金】：北京市自然基金面上项目(7112029)
【分类号】：R739.86

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