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

doi:10.12092/j.issn.1009-2501.2018.03.002

中国临床药理学与治疗学 ›› 2018, Vol. 23 ›› Issue (3): 247-254.doi: 10.12092/j.issn.1009-2501.2018.03.002

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

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

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，宜昌 443002，湖北; ²三峡大学肿瘤微环境与免疫治疗湖北省重点实验室，宜昌 443002，湖北

收稿日期:2017-08-22 修回日期:2017-09-29 出版日期:2018-03-26 发布日期:2018-03-28
通讯作者: 贺海波，男，博士，副教授，硕士生导师，研究方向：中药药理学。 Tel: 0717-6397478 E-mail: hjy219@126.com
作者简介:颜为红，女，硕士研究生，研究方向：中药药理学。 Tel: 0717-6397478 E-mail: 984086176@qq.com
基金资助:
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)

Effects of steroidal saponin RCE-4 from Reineckia Carnea (Andr.) Kunth on Ras/Erk and p16/cyclinD1/ CDK4 signaling pathways in the human cervix cancer Caski cells

YAN Weihong¹, ZOU Kun¹, HE Haibo ^1,2, ZHANG Yongfeng¹, LI Xiaomei¹, LI Xiaoqin¹, YANG Xiaojiao¹, WANG Junzhi ¹, DENG Zhangshuang¹

¹Hubei Key Laboratory of Natural Products Research and Development, College of Biological and Pharmaceutical Sciences, China Three Gorges University & Hubei Institute of Tujia medicine, China Three Gorges University, Yichang 443002, Hubei, China;² Hubei Key Laboratory of Tumor and Microenvironment and immunotherapy, China Three Gorges University, Yichang 443002, Hubei, China

Received:2017-08-22 Revised:2017-09-29 Online:2018-03-26 Published:2018-03-28

摘要/Abstract

摘要：

目的: 研究吉祥草中甾体皂苷RCE-4影响Ras/Erk和p16/cyclinD1/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、cyclinD1和CDK4 mRNA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12.33 μmol/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mRNA表达，下调cyclinD1和CDK4的mRNA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/cyclinD1/CDK4信号通路激活有关。

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

Abstract:

AIM: To investigate the effects of steroidal saponin RCE-4 from Reineckia Carnea (Andr.) Kunth on Ras/Erk and p16/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, respectively; 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 phosphorylated protein of Ras/Erk signaling pathway. RESULTS:RCE-4 could significantly inhibit the growth of Caski cells compared with the control group, the IC50 was 12.33 μmol/L in the 24 h, and it increased the percentage of G0/G1 phase cells, decreased the percentage of S phase cells, inhibited the proteins levels of Ras, p-Raf, p-Mek1/2 and p-Erk1/2, decreased the p-Raf:Raf,p-Mek1/2:Mek1/2 and p-Erk1/2:Erk1/2 ratios, up-regulated the mRNA expression of p16, down-regulated the expressions level of cyclinD1 and CDK4. CONCLUSION: RCE-4 could significantly inhibit the proliferation of Caski cells and induce the cells to block in G0/G1 phase, and the mechanism is related to inhibiting the Ras/Erk and p16/cyclinD1/CDK4 signaling pathways.

Key words: RCE-4, cell cycle, Caski cells, Ras/Erk signaling pathway, p16/cyclinD1/CDK4 signaling pathway

中图分类号:

R965.2

颜为红，邹坤，贺海波，张永峰，李小妹，李小琴，杨小姣，汪鋆植，邓张双. 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[J]. 中国临床药理学与治疗学, 2018, 23(3): 247-254.

YAN Weihong, ZOU Kun, HE Haibo, ZHANG Yongfeng, LI Xiaomei, LI Xiaoqin, YANG Xiaojiao, WANG Junzhi, DENG Zhangshuang. Effects of steroidal saponin RCE-4 from Reineckia Carnea (Andr.) Kunth on Ras/Erk and p16/cyclinD1/ CDK4 signaling pathways in the human cervix cancer Caski cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(3): 247-254.

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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/cyclinD1/ CDK4 signaling pathways in the human cervix cancer Caski cells

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