天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (8): 785-790.doi: 10.11958/20210431

• 细胞与分子生物学 •    下一篇

RITA通过ROS/Src/Stat3通路诱导肺鳞癌H226细胞凋亡

区豪杰，孙嘉，李华宇，董超，刘冰△
  

  1. 广东药科大学药学院（邮编510006）
  • 收稿日期:2021-02-22 修回日期:2021-05-26 出版日期:2021-08-15 发布日期:2021-08-19
  • 通讯作者: 刘冰 E-mail:liubing520@gdpu.edu.cn
  • 作者简介:区豪杰（1997），男，硕士在读，主要从事抗肿瘤新药药理学方面研究。E-mail：280218570@qq.com
  • 基金资助:
    国家自然科学基金资助项目（81972821）

RITA induces cell apoptosis of lung squamous carcinoma H226 via the ROS/Src/Stat3 patway

OU Hao-jie, SUN Jia, LI Hua-yu, DONG Chao, LIU Bing△   

  1. School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China
    △Corresponding Author E-mail: liubing520@gdpu.edu.cn
  • Received:2021-02-22 Revised:2021-05-26 Published:2021-08-15 Online:2021-08-19

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摘要： 目的 探究肿瘤凋亡和P53再生化合物（RITA）对肺鳞癌细胞凋亡的影响及作用机制。方法 采用MTT 细胞活力检测法检测不同浓度RITA对肺鳞癌细胞H226和正常肺上皮细胞BEAS-2B增殖的影响，筛选合适的干预 浓度。经0、0.05、0.1和0.2 μmol/L RITA处理后，采用流式细胞术分析H226细胞内活性氧（ROS）产生和细胞凋亡水 平变化。Western blot 检测 P-Src、Src、转录信号转换器和激活因子 3（Stat3）、P-Stat3、Bim、Mcl-1、存活蛋白 （Survivin）、Bax及B淋巴细胞瘤-2（Bcl-2）的蛋白表达水平。H226细胞经0.2 μmol/L RITA 和5.0 mmol/L 抗氧化剂 N-乙酰半胱氨酸（NAC）同时处理后，观察NAC能否逆转RITA对H226细胞的生物学效应。结果 0.05~0.2 μmol/L 范围内，随着RITA浓度的升高，H226细胞增殖活性下降，半抑制浓度（IC50）为（0.130±0.008）μmol/L，而相同给药浓 度下对 BEAS-2B 细胞增殖无明显影响。0~0.2 μmol/L RITA 可增加 H226 细胞内 ROS 水平并诱导细胞凋亡，下调 P-Src、P-Stat3、Mcl-1、Survivin、Bcl-2蛋白表达，上调Bim、Bax蛋白表达。经NAC处理后，RITA抑制Src/Stat3通路活 性及诱导H226细胞凋亡的效应被逆转。结论 RITA通过提高肺鳞癌H226细胞内ROS的水平，从而抑制Src/Stat3 通路活性，最终诱导细胞凋亡。

关键词: 肺肿瘤, 癌, 鳞状细胞, 活性氧, 基因, P53, 细胞凋亡, STAT3转录因子, 肿瘤凋亡和P53再生化合物, src
族激酶类

Abstract: Objective To explore the effects of reactivator of P53 and induction of tumor apoptosis (RITA) on the cell apoptosis of lung squamous carcinoma cells and its mechanism. Methods MTT assay was used to detect the effects of different concentrations of RITA on the proliferation of H226 and BEAS-2B cells, and the appropriate intervention concentration was screened. After treatment with 0, 0.05, 0.1 and 0.2 μmol/L RITA, reactive oxygen species (ROS) content and apoptosis rate of H226 cells were detected by flow cytometry. The expression levels of P-Src, Src, P-Stat3, Stat3, Bim, Mcl-1, Survivin, Bax and Bcl-2 in H226 cells were detected by Western blot assay. After H226 cells were treated with 0.2 μmol/L RITA and 5.0 mmol/L antioxidant N-acetylcysteine (NAC) at the same time, it was observed whether NAC could reverse the biological effects of RITA on H226 cells. Results With the increment on concentration from 0.05 to 0.2 μmol/L, the viability activity decreased with the increased concentration of RITA. The half inhibitory concentration (IC50) in H226 cells was (0.130±0.008) μmol/L. There was no significant effect on the proliferation of BEAS-2B cells at the same concentration. RITA 0-0.2 μmol/L increased the ROS level and induced apoptosis in H226 cells. Meanwhile, 0-0.2 μmol/L RITA also down-regulated the protein expressions of P-Src, P-Stat3, Mcl-1, Survivin and Bcl-2, while up-regulated Bim and Bax expression. After NAC treatment, the effect of RITA on inhibiting Src/Stat3 pathway activity and inducing apoptosis was reversed in H226 cells. Conclusion RITA inhibits the activity of Src/Stat3 pathway by increasing ROS generation, and thus induces apoptosis of H226 cells.

Key words: lung neoplasms, carcinoma, squamous cell, reactive oxygen species, genes, P53, apoptosis, STAT3 transcription factor, RITA, src-family kinases

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