吉祥草中甾体皂苷RCE-4对人宫颈癌Caski细胞Ras/Erk和p16/cyclinD1/CDK4通路的影响  

Effects of steroidal saponin RCE-4 from Reineckia Carnea(Andr.)Kunth on Ras/Erk and p16/cyclin D1/CDK4 signaling pathways in the human cervix cancer Caski cells

作　　者：颜为红[1] 邹坤[1] 贺海波[1,2] 张永峰[1] 李小妹[1] 李小琴[1] 杨小姣[1] 汪鋆植[1] 邓张双[1] YAN Weihong;ZOU Kun;HE Haibo;ZHANG Yongfeng;LI Xiaomei;LI Xiaoqin;YANG Xiaojiao;WANG Junzhi;DENG Zhangshuang(Hubei Key Laboratory of Natural Products Research and Development, College of Biological and Pharmaceuti- cal Sciences, China Three Gorges University ＆ Hubei Institute of Tujia medicine, China Three Gorges University, Yichang 443002, Hubei, China;Hubei Key Laboratory of Tumor and Microenvironment and immunotherapy, China Three Gorges University, Yichang 443002, Hubei, China)

机构地区：[1]三峡大学天然产物研究与利用湖北省重点实验室&湖北省土家医药研究所 [2]三峡大学肿瘤微环境与免疫治疗湖北省重点实验室

出　　处：《中国临床药理学与治疗学》2018年第3期247-254,共8页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家自然科学基金(21272136);肿瘤微环境与免疫治疗湖北省重点实验室开放基金(2015KZL03)

摘　　要：目的:研究吉祥草中甾体皂苷RCE-4影响Ras/Erk和p16/cyclin D1/CDK4信号通路促进Caski细胞凋亡的作用机制。方法:体外培养人宫颈癌Caski细胞株,用RCE-4作为处理因素,MTT法检测细胞增殖活力;流式细胞术检测细胞周期分布;Real-time PCR检测p16、cyclin D1和CDK4m RNA表达水平;Western blot检测Ras/Erk信号通路总蛋白和磷酸化蛋白表达水平。结果:RCE-4呈剂量依赖性抑制Caski细胞增殖,其作用24 h的IC50值为12.33μmol/L;升高G0/G1期细胞比例,降低S期细胞比例;抑制Ras、p-Raf、p-Mek1/2和p-Erk1/2蛋白表达,降低p-Raf/Raf、p-Mek1/2/Mek1/2和p-Erk1/2/Erk1/2比值,上调p16的m RNA表达,下调cyclin D1和CDK4的m RNA表达。结论:RCE-4可抑制Caski细胞的增殖,诱导细胞在G0/G1期阻滞,其诱导作用与其抑制Ras/Erk和p16/cyclin D1/CDK4信号通路激活有关。AIM ： To steroidal saponin RCE-4 investigate the effects of from Reineckia Carnea （Andr.） Kunth on Ras/Erk and pl6/cyclinD1/ CDK4 signaling pathways in the human cervix cancer Caski cells, and to explore its possible mechanism. METHODS： The Caski cell line was cultured in vitro, the cell viability and the distribution of cell cycle was detected by MTT and flow cytometry, re- spectively; the quantitative real-time PCR was used to evaluate the mRNA expressions of p16, cell cycle-related cyclinD1 and CDK4; Western blot was used to determine the total protein and phosphoryla- ted protein of Ras/Erk signaling pathway. RE- SULTS：RCE-4 could significantly inhibit the growth of Caski cells compared with the control group, the ICso was 12.33 Ixmol/L in the 24 h, and it in- creased the percentage of G0/G1 phase cells, de-creased the percentage of S phase cells, inhibited the proteins levels of Ras, p-Raf, p-Mekl/2 and p- Erkl/2, decreased the p-Raf： Raf, p-Mekl/2： Mekl/2 and p-Erkl/2 ： Erkl/2 ratios, up-regulated the mRNA expression of p16, down-regulated the expressions level of cyclinD1 and CDK4. CON- CLUSION： RCE-4 could significantly inhibit the proliferation of Caski cells and induce the cells to block in Go/G1 phase, and the mechanism is related to inhibiting the Ras/Erk and pl6/cyclinD1/CDK4 signaling pathways.

关 键 词：RCE-4 细胞周期 CASKI细胞 Ras/Erk信号通路 p16/cyclinD1/CDK4信号通路 

分 类 号：R965.2[医药卫生—药理学]