Effects of hammerhad ribozyme targeting connective tissue growth factor on collagen Ⅰ synthesis and cell cycle progression of human hepatic stellate cells / 吉林大学学报(医学版)

ABSTRACT

Objective To study the inhibitory effect of hammerhead ribozyme targeting connective tissue growth factor(CTGF) on collagen I synthesis and cell cycle progression of human hepatic stellate cell line(LX-2) cells.Methods Hammerhead ribozyme cDNA targeting CTGF mRNA plus two self-cleaving sequences were inserted into pTriEx2 vector to construct a recombinant vector pTriCTGF-Rz.LX-2 cells were transfected with either pTriEx2 or pTriCTGF-Rz and further stimulated with or without TGF-1.There were five groups in the experiment:control group,pTriEx2 group,pTriCTGF-Rz group,pTriEx2 plus TGF-?1 group,and pTrCTGF-Rz plus TGF?1 group.Semi-quantitative RT-PCR was used to detect the levels of CTGF mRNA and collagen Ⅰ mRNA.ELISA and flow cytometry were used to detect the levels of collagen Ⅰ secretion and cell cycle.Results Transfection of pTriCTGF-Rz into LX-2 cells reduced the CTGF mRNA and collagen Ⅰ mRNA levels as well as collagen Ⅰ protein level compared with pTriEx2 group(P