国际肿瘤学杂志

国际肿瘤学杂志 ›› 2018, Vol. 45 ›› Issue (11): 647-651.doi: 10.3760/cma.j.issn.1673-422X.2018.11.002

• 论著 • 上一篇    下一篇

微小RNA-134调控P53对非小细胞肺癌细胞增殖及凋亡的影响

沈庆林，宋启斌，章必成，姚颐，徐唐鹏，褚玉新，彭敏   

  1. 430060 武汉大学人民医院肿瘤中心
  • 出版日期:2018-11-08 发布日期:2018-12-21
  • 通讯作者: 彭敏，Email: mpeng320@whu.edu.cn E-mail:mpeng320@whu.edu.cn
  • 基金资助:
    国家自然科学基金（81472748）

Effects of microRNA-134 on proliferation and apoptosis of non-small cell lung cancer by regulating P53 protein

Shen Qinglin, Song Qibin, Zhang Bicheng, Yao Yi, Xu Tangpeng, Chu Yuxin, Peng Min   

  1. Cancer Center, Renmin Hospital of Wuhan University, Wuhan 430060, China
  • Online:2018-11-08 Published:2018-12-21
  • Contact: Peng Min, Email: mpeng320@whu.edu.cn E-mail:mpeng320@whu.edu.cn
  • Supported by:
    National Natural Science Foundation of China (81472748)

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摘要： 目的 探讨微小RNA-134（miR-134）对非小细胞肺癌（NSCLC）细胞增殖和凋亡的影响及其潜在的分子机制。方法 采用实时荧光定量PCR（qRT-PCR）检测miR-134在10对NSCLC肿瘤组织及其癌旁组织之间、正常肺上皮细胞株BEAS-2B与肺腺癌细胞株A549之间的表达差异。A549细胞分别转染miR-NC和miR-134 mimic，采用四甲基偶氮唑蓝（MTT）和集落形成实验检测过表达miR-134对A549细胞株增殖能力的影响；采用流式细胞术检测过表达miR-134对A549细胞凋亡的影响；Western blotting检测过表达miR-134后P53蛋白表达的情况。结果 miR-134在NSCLC肿瘤组织和癌旁组织中的相对表达量分别为0.429±0.126和0.971±0.183，差异有统计学意义(t=7.742，P<0.001)；miR-134在BEAS-2B和A549细胞株中的相对表达量分别为1.013±0.095和0.371±0.068，差异有统计学意义(t=17.377，P<0.001)。A549细胞转染miR-134 mimic后第3、4天的A值分别为0.451±0.051、0.518±0.074，转染miR-NC后第3、4天的A值分别为0.683±0.041、0.815±0.065，miR-134 mimic组细胞增殖能力显著低于miR-NC组（t=12.965，P<0.001；t=9.535，P<0.001）；转染miR-NC和miR-134 mimic后A549细胞集落形成率分别为91.2%±8.3%和38.6%±4.5%，转染miR-134 mimic后A549细胞集落形成率明显降低（t=17.617，P<0.001）；miR-134 mimic组和miR-NC组细胞凋亡率分别93.5%±3.7%和85.4%±2.0%，差异有统计学意义（t=6.119，P<0.001）；P53蛋白在miR-134 mimic组和miR-NC组中的相对表达量分别为1.816±0.173和0.992±0.096，差异有统计学意义(t=19.308，P<0.001)。结论 miR-134可通过提高P53蛋白的表达抑制肿瘤细胞的活性及增殖能力，促进肿瘤细胞凋亡，可能成为治疗NSCLC的有效靶点。

关键词: 癌, 非小细胞肺, 细胞增殖, 细胞凋亡, 肿瘤抑制蛋白质P53, 微小RNA-134

Abstract: Objective To investigate the effects of microRNA-134 (miR-134) on the proliferation and apoptosis of non-small cell lung cancer (NSCLC) and its potential molecular mechanism. Methods Quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) was used to detect the differences of miR-134 expression between 10 cases of lung cancer tissues and normal lung tissues, and between normal human lung epithelial cell line BEAS-2B and lung adenocarcinoma cell line A549. miR-NC and miR-134 mimic were transfected into A549 cells. The effect of miR-134 on proliferation of A549 cells was detected by methyl thiazolyl tetrazolium (MTT) and colony form experiment. Flow cytometry was used to determine the effect of miR-134 on A549 cells apoptosis. The effect of miR-134 on the expression of P53 protein was detected by Western blotting. Results The relative expressions of miR-134 in NSCLC tumor tissues and adjacent tissues were 0.429±0.126 and 0.971±0.183 respectively, and the difference was statistically significant (t=7.742, P<0.001). The relative expressions of miR-134 in BEAS-2B cells and A549 cells were 1.013±0.095 and 0.371±0.068 respectively, and the difference was statistically significant (t=17.377, P<0.001). The absorbance (A) values of A549 cells transfected with miR-mimic were 0.451±0.051 and 0.518±0.074 on the third and forth day respectively, and those of A549 cells transfected with miR-NC were 0.683±0.041 and 0.815±0.065 respectively. The proliferation ability of miR-mimic group was significantly lower than that of miR-NC group (t=12.965, P<0.001; t=9.535, P<0.001). The colony forming rates of A549 cells transfected with miR-NC and miR-134 mimic were 91.2%±8.3% and 38.6%±4.5% respectively, and the colony forming rate of A549 cells in miR-134 mimic group was significantly decreased (t=17.617, P<0.001). The apoptosis rates of miR-134 mimic group and miR-NC group were 93.5%±3.7% and 85.4%±2.0% respectively, and the difference was significant difference (t=6.119, P<0.001). The relative expressions of P53 protein in miR-134 mimic group and miR-NC group were 1.816±0.173 and 0.992±0.096 respectively, and the diffe-rence was statistically significant (t=19.308, P<0.001). Conclusion miR-134 can be an effective target for the treatment of NSCLC by increasing the protein expression of P53, inhibiting the viability and proliferation of tumor cells, and promoting the apoptosis of tumor cells.

Key words: Carcinoma, non-small-cell lung, Cell proliferation, Apoptosis, Tumor suppressor protein P53, MicroRNA-134