摘要
目的观察槐耳清膏对人胃癌细胞NCI-N87细胞自噬的影响。方法以NCI-N87细胞为研究对象,用不同浓度的槐耳清膏(4、8、16、32mg/ml)分别处理细胞24h、48h、72h后,用CCK-8法检测NCI-N87细胞的增殖抑制率,用流式细胞仪检测细胞凋亡及细胞自噬小体含量,并通过RT-PCR和Western blot对自噬相关蛋白包括Beclin-1、LC3-II和PTEN的mRNA和蛋白水平进行检测。结果和对照组相比较,4mg/ml以上浓度的槐耳清膏对细胞的增殖、凋亡及细胞内自噬小体含量均有显著影响,并表现与浓度和时间的正相关性。RT-PCR和Western blot结果显示,与对照组相比,Beclin-1、LC3II和PTEN明显升高。结论槐耳清膏可诱导胃癌细胞NCI-N87自噬。
Objective To investigate the effects of Huaier extract on cell growth and autophagy in gastric caner NCI-N87 cell line.Methods NCI-N87 cells were cultured in vitro and incubated with different concentrations(4、8、16、32mg/ml)last for different time(24,48,72h).CCK-8 assay was developed to detect the cell proliferation inhibition rate after 24,48,72h respectively.The cell apoptotic rate and the content of acidic vesicular organelles content were measured by flow cytometry after 24h.The expressions of autophagy-associated protein Beclin 1,phosphatase and tensin homolog(PTEN),LC3-II were detected by real-time RT-PCR and western blot.Results Huaier extract had significant effect on proliferation,apoptotic rate and acidic vesicular organelles of NCI-N87 cells compared to control group(P>0.05),and showed a dose-and time-dependent manner at the concentration above 4mg/ml(P<0.05).The results of real-time RT-PCR and westem blot showed that compared with control group,the expressions of Beclin I,PTEN and LC3-II were significantly up-regulated(P<0.05).Conclusion Huaier extract induces the auophagy of human gastric cancer NCI-N87 cells.
作者
廖传文
李洁兰
殷俊翔
胡淑琴
饶雪峰
LIAO Chuanwen;LI Jielan;YIN Junxiang(The Department of General Surgery,People’s Hospital of Jiangxi Province,Nanchang,Jiangxi,China 330006;People’s Hospital of Jishui county,Jishui,Jiangxi,331600,China)
出处
《江西医药》
CAS
2018年第11期1232-1235,1258,共5页
Jiangxi Medical Journal
基金
江西省卫生计生委中医药课题
编号2016B083
关键词
槐耳
自噬
胃癌
Autophagy
Huaier extract
Gastric cancer
