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浙江大学学报(理学版)  2017, Vol. 44 Issue (4): 456-463    DOI: 10.3785/j.issn.1008-9497.2017.04.011
生命科学     
穿心莲内酯诱导胃癌细胞周期抑制和内源性凋亡
刘奇章, 龚兴国
浙江大学 生命科学学院 生物化学研究所, 浙江 杭州 310058
Andrographolide induces cycle arrest at G2/M phase and intrinsic apoptosis in gastric carcinoma cells
LIU Qizhang, GONG Xingguo
Institute of Biochemistry, College of life Sciences, Zhejiang University, Hangzhou 310058, China
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摘要: 研究了穿心莲内酯诱导胃癌细胞SGC-7901生长抑制、周期阻滞与细胞凋亡的作用机制.用梯度浓度的穿心莲内酯作用于SGC-7901细胞,以MTT法检测药物对细胞的生长抑制情况;以流式细胞技术检测SGC-7901的周期分布状况、胞内的ROS水平以及线粒体膜电位的变化;以Annexin V-FITC/PI双染法检测细胞凋亡的水平;以Western blot检测不同加药浓度处理后的细胞内相关蛋白水平的变化.可知穿心莲内酯对SGC-7901细胞具有时间依赖和浓度依赖的抑制效果,48 h的IC50约为24 μg· mL-1;并且能够有效抑制G2/M期的周期进展,随后诱发内源性凋亡途径;与此同时,细胞内的ROS水平上升,线粒体膜电位下降;再加入NADPH之后,细胞内的GSH水平有所回升,周期阻滞得到缓解.细胞内的p53、p-CDC2、cleaved caspase-3/9、Bax等上调;bcl-2等水平下调.穿心莲内酯是通过上调胞内ROS水平并使线粒体膜电位崩溃而诱发G2/M期的周期阻滞与内源性凋亡,最终抑制SGC-7901细胞的生长.
关键词: 穿心莲内酯胃癌凋亡诱导周期阻滞    
Abstract: Andrographolide is one of the extractions from Andrographis paniculata, which is a traditional herb early reported its anti-tumor efficacy, and is proved it's the active ingredient which could suppress multiple carcinoma. It is proclaimed that many kinds of mechanisms concentrated on autophagy or apoptosis, and correlative signal pathway. To verify whether andrographolide would prevent amplification of gastric cancer cells, and to investigate the mechanism of andrographolide's effect on cell growth, cycle arrest and induction of apoptosis in vitro, we designed the experiment:SGC7901 was treated with different doses(0, 16, 24 and 32 μg·mL-1) of andrographolide for 24, 48 and 72 h; Cell viability was measured by MTT assays to determine the efficacious time of performance. Then, investigations of cell cycle, ROS level and mitochondria membrane potential were proceeded and detected by flow cytometry. Cell apoptosis was monitored by flow cytometry using Annexin V-FITC/PI staining. Expression level of p53, p-CDC2, bcl-2, Bax were tested by western blot. Finally, we found that andrographolide effectively inhibited SGC-7901 cell growth and the IC50 was 24 μg· mL-1 in 48 h. Meanwhile, the medicine showed the ability of cell cycle arrest at G2/M phase and apoptosis above 60% when treated with 24 μg· mL-1 andrographolide for 48 h. Western blot indicated that cell cycle and apoptosis relevant proteins, such as p53, p-CDC2 and Bax were up-regulated and bcl-2 was down-regulated. In the end, we conclude that andrographolide inhibits SGC-7901 cell proliferation through G2/M phase arrest and inducing intrinsic apoptosis pathway.
Key words: andrographolide    gastric carcinoma    induction of apoptosis    cycle arrest
收稿日期: 2016-11-10 出版日期: 2017-12-09
CLC:  Q50  
通讯作者: 龚兴国,ORCID:http://orcid.org/0000-0003-1206-5154,E-mail:gongxg@zju.edu.cn.     E-mail: gongxg@zju.edu.cn
作者简介: 刘奇章(1990-),ORCID:http://orcid.org/0000-0002-4043-1783,男,硕士,主要从事生物化学与分子生物学研究,E-mail:18268139991@163.com.
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引用本文:

刘奇章, 龚兴国. 穿心莲内酯诱导胃癌细胞周期抑制和内源性凋亡[J]. 浙江大学学报(理学版), 2017, 44(4): 456-463.

LIU Qizhang, GONG Xingguo. Andrographolide induces cycle arrest at G2/M phase and intrinsic apoptosis in gastric carcinoma cells. Journal of ZheJIang University(Science Edition), 2017, 44(4): 456-463.

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https://www.zjujournals.com/sci/CN/10.3785/j.issn.1008-9497.2017.04.011        https://www.zjujournals.com/sci/CN/Y2017/V44/I4/456

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