miR-122表达载体的构建及其对HepG2.2.15细胞增殖的抑制作用

山东大学学报(医学版) ›› 2011, Vol. 49 ›› Issue (10): 118-.

miR-122表达载体的构建及其对HepG2.2.15细胞增殖的抑制作用

范春光1，王春梅2，王燕1，李丽1，孙汶生3，刘玉刚1，李瑞峰1

山东大学医学院 1.病理生理学研究所； 2.微生物研究所； 3.免疫研究所，济南 250012

收稿日期:2011-03-31 出版日期:2011-10-10 发布日期:2011-10-10
通讯作者: 李瑞峰(1955- )，男，教授，主要从事2型糖尿病、胰岛素抵抗发病原因与机制以及衰老与抗衰老机制研究。 E-mail：ruifeng@sdu.edu.cn 刘玉刚(1974- )，男，副教授，主要从事HBV感染及其相关肝癌发生机制的研究。
作者简介:范春光(1985- )，男，硕士研究生，主要从事肝癌发生机制的研究。
基金资助:
国家自然科学基金青年基金资助项目(No.30801036)

Construction of an miR-122 expression vector and its inhibitory effects on proliferation of HepG2.2.15 cells

FAN Chun-guang1, WANG Chun-mei2, WANG Yan1, LI Li1, SUN Wen-sheng3, LIU Yu-gang1, LI Rui-feng1

1. Department of Pathophysiology； 2. Department of Microbiology； 3. Institute of Immunology，School of Medicine， Shandong University, Jinan 250012, China

Received:2011-03-31 Online:2011-10-10 Published:2011-10-10

摘要/Abstract

摘要：

目的构建miR-122表达载体并检测其表达对HepG2.2.15细胞恶性表型的逆转作用。方法以HepG2.2.15细胞基因组DNA为模板，PCR扩增miR-122前体cDNA序列，以质粒pSilencer3.1-H1 neo为母本，构建miR-122表达载体将其转染HepG2.2.15细胞。表达后,利用ELISA检测HepG2.2.15细胞HBV复制和表达的变化，利用CCK8检测细胞增殖的变化。结果构建的miR-122表达载体可在HepG2.2.15细胞中表达。转染后HepG2.2.15细胞中HBV复制、表达以及细胞增殖均被抑制。结论 miR-122表达载体可在HepG2.2.15细胞中有效表达，其表达可抑制HBV复制、表达以及细胞增殖。

关键词: miR-122；表达载体；肝炎病毒，乙型；癌，肝细胞； HepG2.2.15

Abstract:

Objective To construct an miR-122 expression vector and examine the effect of miR-122 expression in reversing the malignant phenotype of the HepG2.2.15 cell line. Methods Genomic DNA of the HepG2.2.15 cell line was used as a template and the target gene fragment was PCR-amplified. The product was confirmed by enzymatic digestion and sequencing. The vector was transfected into HepG2.2.15 cells. HBsAg and HBeAg in the supernatant from cell cultures were measured, and HBV-DNA level was detected by real-time PCR. Cell proliferation after transfection was monitored. Results The MiR-122 expression vector was expressed in HepG2.2.15 cells，leading to inhibition of HBV replication and expression as well as proliferation of HepG2.2.15 cells. Conclusion MiR-122, expressed by means of an expression vector, inhibits HBV replication and expression. Proliferation of HepG2.2.15 cells was also impaired.

Key words: miR-122; Expression vector; Hepatitis B virus; Carcinoma, hepatocellular; HepG2.2.15

中图分类号:

R363.2

范春光1，王春梅2，王燕1，李丽1，孙汶生3，刘玉刚1，李瑞峰1. miR-122表达载体的构建及其对HepG2.2.15细胞增殖的抑制作用[J]. 山东大学学报(医学版), 2011, 49(10): 118-.

FAN Chun-guang1, WANG Chun-mei2, WANG Yan1, LI Li1, SUN Wen-sheng3, LIU Yu-gang1, LI Rui-feng1. Construction of an miR-122 expression vector and its inhibitory effects on proliferation of HepG2.2.15 cells[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2011, 49(10): 118-.

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