吉祥草甾体皂苷RCE-4诱导人宫颈癌Ca Ski细胞凋亡与自噬相互作用的分子机制  

Molecular mechanism of crosstalk between apoptosis and autophagy in human cervical cancer Ca Ski cells induced by RCE-4,a steroidal saponin isolated from Reineckia carnea

作　　者：游方芳 程凡 邹坤 张晶 陈剑锋 YOU Fang-fang;CHENG Fan;ZOU Kun;ZHANG Jing;CHEN Jian-feng(Hubei Key Laboratory of Natural Products Research and Development, College of Biological and Pharmaceutical Sciences, China Three Gorges University, Yichang,Hubei 443002, China)

机构地区：[1]三峡大学生物与制药学院天然产物研究与利用湖北省重点实验室,湖北宜昌443002

出　　处：《中国药理学通报》2021年第12期1727-1735,共9页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81773952)。

摘　　要：目的解析RCE-4诱导的凋亡和自噬在其抑制人宫颈癌Ca Ski细胞增殖中的作用并探索增强其抗肿瘤活性的方法。方法应用各种自噬和凋亡抑制剂、siRNA质粒作为工具,MTT法检测细胞活力,流式细胞术检测细胞凋亡率,Western blot分析凋亡和自噬相关蛋白的表达。结果自噬抑制剂Baf A1与RCE-4协同效果最佳,联用后RCE-4对Ca Ski细胞的半数抑制浓度IC_(50)由4.669μmol·L^(-1)减小到1.368μmol·L^(-1),细胞凋亡率由49.96%增长到91.08%。Beclin 1 siRNA、Bcl-2 siRNA与RCE-4联用可使IC_(50)由4.669μmol·L^(-1)分别减小到1.536和1.582μmol·L^(-1)。凋亡抑制剂Z-VAD-FMK不影响RCE-4对Ca Ski细胞的敏感性。结论凋亡在RCE-4抑制Ca Ski细胞增殖中起主导地位,而自噬则起着保护细胞、拮抗凋亡的作用。可通过联用Baf A1或者抑制Beclin 1、Bcl-2表达来增强Ca Ski细胞对RCE-4的敏感性。Aim To reveal the role of RCE-4-induced apoptosis and autophagy in inhibiting the proliferation of Ca Ski cells and explore ways to enhance its anti-tumor activity.Methods Various autophagy inhibitors,apoptosis inhibitors and siRNA plasmids were served as tools combined with RCE-4.Then MTT assay was used to detect cell viability,flow cytometry to test cell apoptosis rate and Western blot to analyze the expression of autophagy and apoptosis related proteins.Results Baf A1 had the best synergistic effect combined with RCE-4 among those autophagy inhibitors.The IC_(50) of RCE-4 in combination with Baf A1 decreased from 4.669μmol·L^(-1) to 1.368μmol·L^(-1),and the apoptotic rate of Ca Ski cells increased from 49.96%to 91.08%.RCE-4 combined with Beclin 1 siRNA or Bcl-2 siRNA could reduce its IC_(50) from 4.669μmol·L^(-1) to 1.536 or 1.582μmol·L^(-1) respectively.The apoptosis inhibitor Z-VAD-FMK had less obvious effect on increasing the sensitivity of RCE-4 to Ca Ski cells.Conclusion Apoptosis plays a dominant role in the proliferation of Ca Ski cells inhibited by RCE-4,while autophagy plays a role in protecting cells and antagonizing apoptosis.The sensitivity of RCE-4 to Ca Ski cells can be enhanced by the combination with Baf A1 or inhibiting the expression of Beclin 1 and Bcl-2.

关 键 词：RCE-4 凋亡 自噬 相互作用 凋亡和自噬抑制剂 SIRNA质粒 

分 类 号：R284.1[医药卫生—中药学] R329.24[医药卫生—中医学]