人肝脏特异性miR-122表达载体的构建及鉴定

Construction and identification of the human liver-specific miR-122 expression vector

摘要:

人肝脏特异性miRNA-122是肝脏中表达丰度最高的miRNA。为研究该miR-122的生物学功能, 从HepG2细胞基因组中用PCR的方法扩增了miR-122的前体, 构建了miR-122的表达载体pLMP-miR-122。pLMP-miR-122质粒转染人正常肝细胞系L-O2和肝癌细胞系HepG2后, 细胞内成熟miRNA-122的表达量显著增加。该质粒与HBV1.3共转染HepG2细胞72 h后, HBV的HBs和HBe蛋白水平的表达量均下降, 说明miRNA-122参与了HBV基因的复制和表达的调控, 为进一步研究miRNA-122的功能和其他一些肝病如HCC的调控机制打下基础。

Abstract:

miR-122 is the most abundant miRNA in adult human liver. To study the functions of miR-122 in liver disease, we amplified the precursor of human miR-122 gene by polymerase chain reaction (PCR) from the HepG2 genomic DNA, and then constructed miR-122 expression vector pLMP-miR-122. pLMP-miR-122 could overexpress mature miR-122 when human normal liver cells L-O2 and the hepatoma cells HepG2 were transfected with it. When HepG2 cells were co-transfected with HBV1.3 and pLMP-miR-122, we found that miR-122 could down-regulate the expression of HBs and HBe antigen. These results showed that the human liver specific miR-122 expression vector was constructed successfully, and it could regulate the replication and the expression of HBV genes. The plasmid pLMP-miR-122 will facilitate further studies of the functions of miR-122 in the development of liver virus infection diseases and HCC.