| 常 远,谭立君,王启威.MiR-125a靶向调节P53表达对人喉癌细胞增殖、凋亡以及放射治疗敏感性的影响[J].中国肿瘤,2020,29(9):708-714. |
| MiR-125a靶向调节P53表达对人喉癌细胞增殖、凋亡以及放射治疗敏感性的影响 |
| Effect of MiR-125a on Proliferation，Apoptosis and Radiosensitivity of Human Laryngeal Cancer Cells and Its Mechanism |
| 中文关键词 修订日期：2020-02-05 |
| DOI：10.11735/j.issn.1004-0242.2020.09.A012 |
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| 中文关键词: miR-125a p53 增殖 凋亡 放射敏感性 喉癌 |
| 英文关键词:miR-125a p53 proliferation apoptosis radiosensitivity laryngeal carcinoma |
| 基金项目:黑龙江省自然科学基金（H2017033） |
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| 中文摘要: |
| 摘 要：［目的］ 探究miR-125a通过靶向调节p53的表达对人喉癌Hep-2细胞增殖、凋亡以及放射治疗敏感性的影响。［方法］ 培养人喉癌Hep-2细胞系，利用慢病毒分别转染miR-125a与空白质粒，RT-PCR法检测转染后各组细胞中miR-125a水平，采用MTT法检测细胞增殖情况。X线照射各组细胞后，行克隆形成实验、微核检测和流式细胞检测法检测转染细胞凋亡水平及放射敏感性，通过在线软件预测p53基因为miR-125a直接调控的靶基因，Western blot检测凋亡相关蛋白水平并进一步验证miR-125a与p53的关系，及在喉癌细胞放射治疗中的作用。［结果］ MTT实验结果示，转染miR-125a后Hep-2细胞增殖能力明显下降（P<0.05）。克隆形成实验与微核检测结果表明高表达miR-125a可使Hep-2细胞放射敏感性显著提高。流式细胞仪检测结果0Gy下Hep-2-miR-125a、Hep-2-con220组，10Gy下Hep-2-miR-125a、Hep-2-con220组凋亡率分别为(6.30%±0.11%)，(0.09%±0.02%)，(18.15%±2.49%)，(1.95%±0.08%)(P<0.01)。TargetScan软件预测TRIAP1（TP53 regulated inhibitor of apoptosis 1）为miR-125a的直接调控的靶基因，Western blot实验结果显示高表达miR-125a可正向调控p53的表达，从而增强Hep-2细胞对X线的放射敏感性。［结论］ MiR-125a表达上调后可能通过上调p53的表达使Hep-2细胞增殖受到抑制，诱使其发生凋亡，并提高放射敏感性。 |
| 英文摘要: |
| Abstract：［Purpose］ To investigate the effect of miR-125a on the proliferation，apoptosis and radiosensitivity of human laryngeal carcinoma cells and its relation with p53 expression. ［Methods］ Human laryngeal carcinoma Hep-2 cells were cultured and transfected with miR-125a and blank plasmids respectively by lentivirus. The level of miR-125a in each group of cells after transfection was detected by RT-PCR，and the cell proliferation was detected with MTT. After X-ray irradiation，the level of apoptosis and radiosensitivity of transfected cells were detected by clone formation test，micronucleus test and flow cytometry. P53 gene was predicted as a directly regulated target gene of miR-125a by online software. The level of apoptosis related proteins was detected，and the relationship between miR-125a and p53 and the role of miR-125a in radiotherapy of laryngeal cancer cells were verified by Western blot method. ［Results］ MTT assay showed that the cell proliferation capacity of Hep-2 cells decreased significantly after miR-125a transfection(P<0.05). The results of clone formation test and micronucleus test showed that overexpression of miR-125a significantly increased the radiosensitivity of Hep-2 cells. The results of flow cytometry showed that the apoptotic rates of Hep-2-miR-125a and Hep-2-con220 groups were (6.30%±0.11%) and (0.09%±0.02%) at 0Gy ，and (18.15%±2.49%) and (1.95%±0.08%) at 10Gy，respectively(P<0.01). TP53 regulated inhibitor of apoptosis 1(TRIAP1) was predicted to be the target gene of miR-125a by TargetsSan software. Western blotting showed that high expression of miR-125a positively regulated the expression of p53，and enhanced the radiosensitivity of Hep-2 cells to X-ray. ［Conclusion］ Overexpression of miR-125a might inhibit Hep-2 cell proliferation，induce apoptosis and enhance its radiosensitivity by up-regulating p53 expression. |
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