Inhibitory Effects of shRNA on Taurine Transporter of Rats Cardiac Myoblasts

YANG Qunhui; YANG Jiancheng; LIU Mei; WU Gaofeng; FENG Ying; HU Jianmin

doi:10.3969/j.issn.1006-267x.2013.11.017

Chinese Journal of Animal Nutrition >

2013 , Vol. 25 >Issue 11: 2643 - 2648

DOI: https://doi.org/10.3969/j.issn.1006-267x.2013.11.017

Molecular Nutrition

Inhibitory Effects of shRNA on Taurine Transporter of Rats Cardiac Myoblasts

YANG Qunhui ,
YANG Jiancheng ,
LIU Mei ,
WU Gaofeng ,
FENG Ying ,
HU Jianmin

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College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110866, China

Received date: 2013-05-13

Online published: 2013-10-27

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Abstract

This experiment was conducted to investigate the inhibitory effects of short hairpin RNA (shRNA) on taurine transporter (TauT) of rats cardiac myoblasts. Three shRNAs expression vectors (shRNA1#, shRNA2# and shRNA3#) and one negative control shRNA expression vector were constructed to be targeted directly at TauT gene. Then the recombinant plasmids shRNA1#, shRNA2#, shRNA3# and shRNA-Neg were transfected into H9c2 cells with liposomes Lipofectamine^TM 2000, the cells were divided into control group (untransfected group), shRNA-Neg group (negative control group), and three transfected groups (shRNA1# group, shRNA2# group and shRNA3# group), each group contained three replicates. After transfection, TauT mRNA expression level was examined by using real-time quantitative PCR, and the state of cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. The results showed that compared with the negative control group and untransfected group, the shRNA1# plasmid significantly decreased the TauT mRNA expression level at 24 h after transfection (P<0.01), the shRNA2# plasmid significantly decreased the TauT mRNA expression level at 24, 48 and 72 h after transfection (P<0.01), and the shRNA3# plasmid significantly decreased the TauT mRNA expression level at 72 h after transfection (P<0.01). The MTT detected result showed that the cell proliferation in the transfected cells was not significantly affected by shRNAs plasmid compared with untransfected cells and negative control cells at 24, 48, 72 h (P>0.05). The present study indicates that the shRNA2# recombinant plasmid can effectively inhibit the expression of TauT gene and does not affect the proliferation of H9c2 cells at 72 h after transfection, which will be of benefit to the further study on the functions of TauT and taurine in cardiocytes metabolism.

Key words： H9c2 cell; RNAi; TauT; cell proliferation

Cite this article

YANG Qunhui , YANG Jiancheng , LIU Mei , WU Gaofeng , FENG Ying , HU Jianmin . Inhibitory Effects of shRNA on Taurine Transporter of Rats Cardiac Myoblasts[J]. Chinese Journal of Animal Nutrition, 2013 , 25(11) : 2643 -2648 . DOI: 10.3969/j.issn.1006-267x.2013.11.017

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