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中国癌症防治杂志

中国癌症防治杂志 ›› 2019, Vol. 11 ›› Issue (4): 308-312.doi: 10.3969/j.issn.1674-5671.2019.04.07

• 基础研究 • 上一篇    下一篇

miR-125b-5p对喉鳞状细胞癌细胞能量代谢及增殖的影响

  

  1. 曹县人民医院耳鼻喉科；山东省肿瘤医院放疗科

  • 出版日期:2019-08-25 发布日期:2019-09-17

Effect of miR-125b-5p on energy metabolism and proliferation of laryngeal squamous cell carcinoma#br#

  • Online:2019-08-25 Published:2019-09-17

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摘要： 目的  探讨miR-125b-5p对喉鳞状细胞癌（laryngeal squamous cell carcinoma，LSCC）细胞能量代谢和增殖的影响及其可能的作用机制。方法 实验分为miR-125b-5p转染组（miR-125b-5p模拟物转染LSCC）和对照组（空质粒转染LSCC细胞）。采用RT-qPCR 检测miR-125b-5p在正常支气管上皮细胞和LSCC细胞中的表达，CCK-8和流式细胞术检测LSCC细胞增殖、凋亡情况，Western blot检测miR-125b-5p过表达后LSCC中HK2的表达，3H-2DG法和乳酸盐比色测定实验分别检测LSCC细胞葡萄糖消耗和乳酸产生的情况。结果 RT-qPCR实验结果显示，与正常支气管上皮细胞相比，LSCC细胞中miR-125b-5p的表达降低（0.68±0.03 vs 0.22±0.05，t=7.025，P=0.001）。CCK-8实验结果显示，转染72 h和96 h后，miR-125b-5p转染组LSCC细胞的增殖能力均较对照组明显降低（P<0.05）。流式细胞术检测结果显示，miR-125b-5p转染组LSCC细胞凋亡率较对照组升高 ［（37.52±2.34）% vs （12.46±3.52）%，t=7.025，P<0.001）］，但细胞集落形成能力降低（0.29±0.02 vs 1.02±0.03，t=5.689，P=0.005）；荧光素酶报告基因测定实验结果显示，miR-125b-5p转染组的荧光素酶活性低于对照组（0.32±0.03 vs 1.01±0.02，t=7.543，P=0.001）；Western blot法实验结果显示，miR-125b-5p转染组HK2蛋白表达水平较对照组降低（0.12±0.02 vs 0.75±0.03，t=5.875，P=0.023）；miR-125b-5p转染组葡萄糖消耗量［（3.85±0.86） dpm/mg vs （10.52±1.34） dpm/mg，t=6.118，P=0.005］以及乳酸产生量［（4.23±1.36） dpm/mg vs （10.96±2.45） dpm/mg，t=5.907，P=0.002］亦较对照组降低。结论 miR-125b-5p可能通过下调HK2表达降低喉鳞状细胞癌细胞的能量代谢和增殖能力，诱导细胞凋亡。

关键词: 喉鳞状细胞癌, miR-125b-5p, 己糖激酶2, 糖酵解, 增殖

Abstract: Objective To investigate the effect of miR-125b-5p affecting energy metabolism and proliferation of laryngeal squamous cell carcinoma by down-regulating hexokinase 2. Methods LSCC cells were divided into miRNA-125b-5p transfection group and control group. The expression of miR-125b-5p in normal bronchial epithelial cell line and LSCC cell line was detected by RT-qPCR，proliferation and apoptosis of LSCC cells were detected by CCK-8 and flow cytometry，HK2 expression in LSCC after overexpression of miR-125b-5p was detected by Western blot，glucose consumption and lactic acid production by LSCC cells were determined by 3H-2DG and lactate colorimetry. Results RT-qPCR results showed that the expression of miR-125b-5p in LSCC cell line was lower than that in normal cell line（0.22±0.05 vs 0.68±0.03，t=7.025，P=0.001）. CCK-8 results showed that after 72 h and 96 h of transfection，the proliferation of LSCC cells in the miR-125b-5p transfection group was lower than that in the control group（P<0.05）. The results of flow cytometry showed that the apoptotic rate of LSCC cells in the miR-125b-5p transfection group was higher than that in the control group［（37.52±2.34）% vs （12.46±3.52）%，t=7.025，P<0.001）］. The colony-forming ability of LSCC cells in the miR-125b-5p transfection group was also lower than that in the control group（0.29±0.02 vs 1.02±0.03，t=5.689，P=0.005）. The results of luciferase reporter gene assay showed that the luciferase activity of the miR-125b?-5p transfection group was lower than that in the control group（0.32±0.03 vs 1.01±0.02，t=7.543，P=0.001）. The results of Western blot showed that the protein level of HK2 in LSCC cells over-expressed by miR-125b-5p in the miR-125b-5p transfection group was lower than that in the control group（0.12±0.02 vs 0.75±0.03，t=5.875，P=0.023）. The results of lactate colorimetriy assay showed that the glucose consumption of LSCC cells in the miR-125b-5p transfection group was lower than that in the control group［（3.85±0.86） dpm/mg vs （10.52±1.34） dpm/mg，t=6.118，P=0.005］. The production of lactic acid in LSCC cells transfected with miR-125b-5p was also lower than that in the control group［（4.23±1.36） dpm/mg vs （10.96±2.45） dpm/mg，t=5.907，P=0.002］. Conclusion miR-125b-5p may reduce the energy metabolism and proliferation of laryngeal squamous cell carcinoma by down-regulating HK2 expression, and induce apoptosis.

Key words: Laryngeal squamous cell carcinoma, miR-125b-5p, Hexokinase 2, Glycolysis, Proliferation

中图分类号: 

  • R739.65