转染miR-124模拟物的宫颈癌细胞株Siha放射敏感性变化及其机制探讨

发布时间：2018-12-19 09:02

【摘要】：目的观察过表达微小RNA124(miR-124)对宫颈癌Siha细胞放射敏感性的影响,并探讨其机制。方法将宫颈癌Siha细胞分为观察组和对照组,观察组转染miR-124模拟物,对照组不转染。采用实时荧光定量PCR法检测两组细胞miR-124和信号转导和转录激活因子3(STAT3)mRNA表达量。采用克隆形成实验检测两组细胞放射敏感性。采用流式细胞术检测两组细胞周期分布和放射后细胞凋亡率。结果观察组、对照组D0分别为1.062、1.542Gy,Dq分别为1.605、1.970 Gy,SF2分别为0.527、0.685。观察组相对于对照组的放射增敏比为1.451。转染后24h观察组、对照组miR-124相对表达量分别为89.3±13.6、1.0±0.1,STAT3 mRNA相对表达量分别为0.3±0.03、1.0±0.1,两组相比,P均0.05。观察组G0/G1期细胞比例为82.49%±1.97%、S期细胞比例11.87%±1.38%、G2/M期细胞比例为5.64%±0.72%;对照组分别为74.58%±1.28%、19.88%±0.26%、5.54%±1.05%。观察组G0/G1期比例高于对照组组,S期比例低于对照组(P均0.05),两组G2/M期细胞比例相比P0.05。观察组、对照组细胞X线照射后细胞凋亡率分别为45.87%±3.16%、37.27%±0.87%,两组相比,P0.05。结论过表达miR-124可能提高宫颈癌Siha细胞的放射敏感性。其机制可能是过表达miR-124会抑制宫颈癌Siha细胞STAT3表达,进而阻滞细胞周期于G0/G1期,促进细胞凋亡,从而提高宫颈癌Siha细胞的放射敏感性。
[Abstract]:Objective to investigate the radiosensitivity of cervical cancer Siha cells induced by overexpression of small RNA124 (miR-124) and its mechanism. Methods Cervical carcinoma Siha cells were divided into observation group and control group. The miR-124 mimics were transfected in the observation group, but not in the control group. The expression of miR-124 and signal transduction and transcription activator 3 (STAT3) mRNA was detected by real-time fluorescence quantitative PCR. The radiosensitivity of two groups of cells was detected by clone forming assay. Cell cycle distribution and apoptosis rate after radiation were detected by flow cytometry. Results in the observation group, the D0 of the control group was 1.062 (1.542Gy) and the DQ was 1.6051.970 Gy,SF2 (0.527) 0.685respectively. The radiosensitization ratio of the observation group was 1.451compared with that of the control group. 24 hours after transfection, the relative expression of miR-124 in the control group was 89.3 卤13.6N 1.0 卤0.1 卤0.1, respectively. The relative expression of STAT3 mRNA in the control group was 0.3 卤0.03 卤1.0 卤0.1, respectively, compared with that in the two groups (P < 0.05). In the observation group, the percentage of cells in G0/G1 phase was 82.49% 卤1.97% and the proportion of cells in S phase was 11.87% 卤1.38% in G _ 2 / M phase, 5.64% 卤0.72% in G _ 2 / M phase. The control group was 74.58% 卤1.28% 卤19.88% 卤0.26% and 5.54% 卤1.05% respectively. The ratio of G0/G1 phase and S phase in the observation group was higher than that in the control group (P 0.05), and the ratio of G 2 / M phase cells in the two groups was lower than that in the control group (P 0.05). In the observation group, the apoptosis rate of the cells in the control group was 45.87% 卤3.16% and 37.27% 卤0.87%, respectively, compared with that in the control group (P 0.05). Conclusion overexpression of miR-124 may enhance the radiosensitivity of cervical cancer Siha cells. The mechanism may be that overexpression of miR-124 can inhibit the expression of STAT3 in cervical cancer Siha cells, thus block cell cycle in G0/G1 phase, promote cell apoptosis, and thus enhance the radiosensitivity of cervical cancer Siha cells.
【作者单位】：蚌埠医学院研究生院;泰兴市人民医院;
【基金】：蚌埠医学院研究生科研创新计划(Byycx1624)
【分类号】：R737.33

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