国际肿瘤学杂志

国际肿瘤学杂志 ›› 2015, Vol. 42 ›› Issue (3): 161-164.doi: 10.3760/cma.j.issn.1673422X.2015.03.001

• 论著 •    下一篇

微小RNA27a对黑色素瘤WM239细胞增殖和凋亡的影响

马亚南, 王宝红, 苏庆红, 许晓群, 王郡甫   

  1. 济南大学山东省医学科学院医学与生命科学学院（马亚南、苏庆红、许晓群、王郡甫）；山东省临沂市中医医院感染管理科（王宝红）
  • 出版日期:2015-03-08 发布日期:2015-01-29
  • 通讯作者: 许晓群，Email： xuxiaoqunsd@163.com E-mail:xuxiaoqunsd@163.com
  • 基金资助:

    山东省自然科学基金（ZR2010CM067）；山东省大型科学仪器设备升级改造技术研究专项(2013SJGZ09）

Effect of miR27a on cell proliferation and apoptosis in human melanoma cell line WM239

Ma Yanan, Wang Baohong, Su Qinghong, Xu Xiaoqun, Wang Junfu   

  1. Institute of Basic Medicine, Shandong Academy of Medical Science; School of Medicine and Life Sciences, University of Ji′nanShandong Academy of Medical Science, Ji′nan 250062, China
  • Online:2015-03-08 Published:2015-01-29
  • Contact: Xu Xiaoqun E-mail:xuxiaoqunsd@163.com

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摘要： 目的探讨微小RNA27a（miR27a）模拟物和抑制物转染黑色素瘤WM239细胞后对细胞增殖和凋亡的影响。方法将miR27a模拟物、抑制物及其阴性对照转染WM239细胞，荧光显微镜观察转染效率，实时荧光定量PCR检测相应的微小RNA，四甲基偶氮唑盐(MTT)法检测细胞增殖，流式细胞仪检测细胞凋亡和细胞周期。结果细胞转染效率为80%～90%。转染miR27a模拟物后，细胞内miR27a表达量明显上升（2△△CT值为26.98±0.01)，与正常对照组相比差异有统计学意义(t＝-1 123.67，P=0.00）；转染miR27a抑制物后，细胞中miR27a的表达量下降（2△△CT值为0.96±0.02)，与正常对照组相比差异无统计学意义(t＝4.04，P＝0.06）。转染miR27a模拟物后，细胞增殖受到明显抑制，与正常对照组相比差异具有统计学意义［72 h吸光度（0.45±0.02）∶（0.72±0.01），F＝129.56，P＜0.05］。miR27a模拟物组G0G1期的细胞比例升高［（74.83±1.46）∶（63.73±1.25），F＝30.33，P＜0.05］，S期和G2M期细胞比例减少［（21.33±1.75）∶（27.50±1.25），F＝14.98，P＜0.05；（3.90±1.31）∶（8.80±2.10），F＝3.66，P＜0.05］；模拟物组细胞凋亡率与正常对照组相比明显增加［（29.67±0.91）%∶（1.44±0.85）%，F＝530.90，P＜0.01］；而抑制物组对细胞周期和凋亡无明显作用。结论miR27a抑制黑色素瘤细胞增殖，具有抑瘤作用，这与其促进细胞凋亡，阻滞细胞周期于G0G1期相关。

关键词: 黑色素瘤, 细胞增殖, 细胞凋亡, 转染, 微小RNA27a

Abstract: ObjectiveTo investigate the effect of miR27a mimic and inhibitor on proliferation and apoptosis in melanoma cell line WM239. MethodsThe miR27a mimic, inhibitor and its negative control were transfected into WM239 cells. The transfection efficiency was evaluated by fluorescence microscope. The expression of miR27a was detected by realtime fluorescent quantitative PCR. The proliferation of cells was detected by MTT. The cell apoptosis and cell cycle were analyzed by flow cytometry. ResultsThe transfection efficiency in WM239 cells was 80% to 90%. The expression of miR27a was markedly upregulated in miR27a mimic group（2△△CT value is 26.98±0.01), with statistically significant difference(t＝-1 123.67, P=0.00）; and the miR27a inhibitor group showed lower expression of miR27a（2△△CT value is 0.96±0.02),  there was no statistically significant difference compared with normal control group(t＝0.04, P＝0.06）. The proliferation of cells was obviously inhibited in miR27a mimic group, and there was statistically significant difference compared with normal control group ［absorbance of 72 h（0.45±0.02）∶（0.72±0.01），F＝129.56, P＜0.05］. The percentage of WM239 cells in G0G1 phase was increased［（74.83±1.46）∶（63.73±1.25），F＝30.33, P＜0.05］, and the percentage of WM239 cells in S phase and G2M phase were decreased ［（21.33±1.75）∶（27.50±1.25），F＝14.98，P＜0.05；（3.90±1.31）∶（8.80±2.10），F＝3.66，P＜0.05］. The apoptosis rate of cells was significantly increased in miR27a mimic group compared with normal group ［（29.67±0.91）%∶（1.44±0.85）%，F＝530.90，P＜0.01］, but the inhibitor group had no obvious effect on cell cycle and cell apoptosis. ConclusionMiR27a can suppress melanoma cell proliferation and act as a tumor suppressor gene, which is relevant to induce cell apoptosis and block cell cycle in G0G1 phase.

Key words: Melanoma, Cell proliferation, Apoptosis, Transfection, microRNA27a