葡萄糖神经酰胺葡萄糖基转移酶siRNA对7702肝细胞增殖的影响及机制研究*

doi:10.3969/j.issn.1672-5069.2018.05.011

实用肝脏病杂志 ›› 2018, Vol. 21 ›› Issue (5): 697-700.doi: 10.3969/j.issn.1672-5069.2018.05.011

葡萄糖神经酰胺葡萄糖基转移酶siRNA对7702肝细胞增殖的影响及机制研究^*

李俊峰, 郑素军, 刘霜, 任锋, 陈煜, 段钟平

730000 兰州市兰州大学第一医院感染病科/传染病学研究室（李俊峰）; 首都医科大学附属北京佑安医院人工肝中心（郑素军,刘霜,陈煜,段钟平）; 北京市肝病研究所（任锋）

收稿日期:2017-08-30 出版日期:2018-09-10 发布日期:2018-09-27
通讯作者: 段钟平,E-mail: duan2517@163.com
作者简介:李俊峰，男，34岁，医学博士，主治医师。主要从事肝脏疾病的基础与临床研究。E-mail：junfenglee@126.com
基金资助:
“十三五” 国家艾滋病和病毒性肝炎等重大传染病防治科技重大专项 (编号：2017ZX10201201/2017ZX10203201-005/2017ZX10202203-006/2017ZX10302201-004); 北京市医院管理局“登峰”人才计划项目（编号：DFL20151601）; 兰州大学第一医院院内基金项目（编号：ldyyyn2017-17）

Impact of UGCG siRNA on 7702 hepatocyte proliferation in vitro

Li Junfeng, Zheng Sujun, Liu Shuang, et al.

Institute of Infectious Diseases,Department of Infectious Diseases,First Hospital,Affiliated to Lanzhou University,Lanzhou 730000,Gansu Province,China

Received:2017-08-30 Online:2018-09-10 Published:2018-09-27

摘要/Abstract

摘要： 目的探讨糖鞘脂合成代谢在肝细胞增殖中的可能作用机制。方法体外培养肝细胞株7702细胞,利用UDP-葡萄糖神经酰胺葡萄糖基转移酶（UGCG）siRNA转染肝细胞。采用MTT法检测细胞增殖,采用实时荧光定量PCR法检测Bcl-2、Bax和Caspase 3基因水平,采用Western blot法检测肝细胞Caspase 3蛋白表达情况。结果 UGCG siRNA干扰组细胞糖化神经酰胺合成酶基因水平比对照组明显下调（P<0.05）;干扰糖化神经酰胺合成酶基因表达后,转染组细胞Bcl-2 mRNA水平显著下降（P<0.05）,Bax mRNA水平显著升高,Caspase 3 mRNA及其蛋白表达水平均上调（P值均<0.05）。结论糖化神经酰胺合成酶的缺乏引起的鞘脂代谢改变可能参与肝细胞的增殖的过程中,这可能与BCL2/BAX调节的细胞信号通路有关。

关键词: 7702肝细胞, 鞘脂, 糖化神经酰胺合成酶, 细胞增殖, 体外

Abstract: Objective To investigate the impact of UGCG siRNA on 7702 hepatocyte proliferation in vitro. Methods The 7702 hepatocytes were cultured in vitro and UDP-glucose ceramide glucosyltransferase（UGCG） siRNA was transfected into the hepatocytes. MTT was performed to detect the cell proliferation,the Bcl-2,Bax,Caspase 3 gene were detected by real-time quantitative PCR,and the expression of Caspase 3 protein in hepatocytes was detected by Western blot. Results UGCG siRNA successfully down-regulated glucosylceramide synthase mRNA levels as compared to that in the control（P<; 0.05）;the proliferation of hepatocytes was inhibited（P<; 0.05） after transfection of the glycosylated ceramide synthase gene,Bcl-2 mRNA decreased（P<; 0.05）,Bax mRNA increased （P<; 0.05）,and the expression of Caspase 3 protein was significantly upregulated（P<; 0.05）. Conclusion The changes of sphingolipid metabolism caused by the lack of glucosylceramide synthase is involved in hepatocyte proliferation, which might be related to the regulation of BCL2/BAX signal pathway.

Key words: 7702 hepatocytes, Sphingolipid, Cell proliferation, Glucosylceramide synthase, In vitro

李俊峰, 郑素军, 刘霜, 任锋, 陈煜, 段钟平. 葡萄糖神经酰胺葡萄糖基转移酶siRNA对7702肝细胞增殖的影响及机制研究^*[J]. 实用肝脏病杂志, 2018, 21(5): 697-700.

Li Junfeng, Zheng Sujun, Liu Shuang, et al.. Impact of UGCG siRNA on 7702 hepatocyte proliferation in vitro[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2018, 21(5): 697-700.

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葡萄糖神经酰胺葡萄糖基转移酶siRNA对7702肝细胞增殖的影响及机制研究^*

Impact of UGCG siRNA on 7702 hepatocyte proliferation in vitro

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