吉祥草活性成分RCE-4触发内质网应激调控人宫颈癌Ca Ski细胞周期阻滞  

Effects of RCE-4,an active component of Reineckia carnea,on regulation of cell cycle arrest in human cervical cancer Ca Ski cells by triggering endoplasmic reticulum stress

作　　者：牟丽云 邢翔飞[1] 陈重旭 张晶 金桂兰[1] 奚伟 陈剑锋 MOU Li-yun;XING Xiang-fei;CHEN Chong-Xu;ZHANG Jing;JIN Gui-lan;XI Wei;CHEN Jian-feng(Dept of Pharmacy,the People’s Hospital of Three Gorges University,the First People’s Hospital of Yichang,Yichang Hubei 443000,China;Medical College of Three Gorges University,Yichang,Hubei 443002,China;Hubei Key Laboratory of Natural Products Research and Development,College of Biological and Pharmaceutical Sciences of China Three Gorges University,Yichang Hubei 443002,China)

机构地区：[1]三峡大学人民医院,宜昌市第一人民医院药学部,湖北宜昌443000 [2]三峡大学医学院,湖北宜昌443002 [3]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9期1224-1231,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81773952);湖北省卫生健康委员会中医药科研项目(No ZY2019M028)。

摘　　要：目的探索吉祥草活性成分RCE-4抑制宫颈癌Ca Ski细胞增殖的机制。方法四甲基偶氮唑蓝法(MTT法)和克隆形成实验考察RCE-4对Ca Ski细胞生长的影响,并计算其半抑制浓度(IC_(50));流式细胞术检测细胞周期的变化;Western blot检测内质网应激及细胞周期相关分子蛋白水平的表达。结果RCE-4对人宫颈癌Ca Ski细胞的增殖具有明显抑制作用,作用24、48、72 h的IC_(50)值分别为8.61、4.69、4.34μmol·L^(-1)。Western blot结果显示RCE-4激活了内质网应激PERK/eIF2α/ATF4/CHOP信号通路。流式细胞术结果显示RCE-4可明显诱导细胞G 2/M期阻滞,细胞内CDK1、cdc25C、p-cdc25C、CyclinB1、Chk2、p-Chk2、GADD45α、MDM2蛋白水平明显降低,p21、p27和ATM蛋白水平明显升高。结论RCE-4可能通过触发内质网应激诱导Ca Ski细胞G 2/M期阻滞从而抑制人宫颈癌Ca Ski细胞增殖。Aim To explore the mechanism of RCE-4,an active constituent isolated from Reineckia carnea on anti-proliferation activity of human cervical neoplasm Ca Ski cells.Methods Etramethylazolyl blue method(MTT)and clone formation assay were adopted to observe the inhibitory effects of RCE-4 on the growth of Ca Ski cells,and its half maximal inhibitory concentration(IC_(50))was calculated.Flow cytometry was used to investigate the distribution of cell cycle of Ca Ski cells after treated with RCE-4.Western blot was used to detect the expression of classical molecular proteins related to endoplasmic reticulum stress(ERS)and cell cycle.Results RCE-4 had an inhibitory effect on the proliferation of cervical cancer Ca Ski cells,with the IC_(50) of 8.61,4.69,4.34μmol·L^(-1) at 24 h,48 h and 72 h respectively.Western blot results showed that RCE-4 activated PERK/eIF2α/ATF4/CHOP signaling pathway related to endoplasmic reticulum stress.Flow cytometry results revealed that RCE-4 could significantly induce G 2/M phase cell cycle arrest,and intracellular CDK1,cdc25C,p-cdc25C,CyclinB1,Chk2,p-Chk2,GADD45α,MDM2 protein levels were significantly reduced,p21,p27 and ATM protein levels increased significantly.Conclusions RCE-4 may induce G 2/M phase arrest of Ca Ski cells through endoplasmic reticulum stress,thereby inhibiting the proliferation of human cervical cancer Ca Ski cells.

关 键 词：吉祥草 RCE-4 宫颈癌 内质网应激 细胞周期阻滞 细胞增殖 

分 类 号：R284.1[医药卫生—中药学] R285.5[医药卫生—中医学]