Effect of Gene Ring Finger Protein 6 on the Expression of Insulin Receptor Substrate-2 in Hepatoma Cells

Abstract

Abstract: It was to construct an eukaryotic expression vector of ring finger protein 6（RNF6）gene and investigate the effect of RNF6 on the expression of insulin receptor substrate-2（IRS-2）. The coding sequence of hRNF6 gene was amplified by PCR with human cDNA as template. The pcDNA3.1-CHA-RNF6 was constructed and transfected into hepatocarcinoma cells（HepG2）by routine molecular biology technology. The total RNA was extracted from HepG2 cells 72 hours post-transfection, the expression levels of IRS-2 was detected by real-time quantitative PCR. Western blotting was applied to detect the protein levels of IRS-2. Result showed that the mRNA level of IRS-2 gene in transfected HepG2 was 37% of the control. The expression level of IRS-2 was lower than the control group significantly （P﹤0.01）. The expression of IRS-2 was down-regulated in HepG2 significantly, and the disorder in insulin signal transduction pathway, which may result from enhanced ubiquitylation level of IRS-2.

Key words: Insulin receptor substrate-2, Ubiquitin, RNF6, Insulin

Gong Jian, Song Jian. Effect of Gene Ring Finger Protein 6 on the Expression of Insulin Receptor Substrate-2 in Hepatoma Cells[J]. Biotechnology Bulletin, 2014, 0(6): 225-228.

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Effect of Gene Ring Finger Protein 6 on the Expression of Insulin Receptor Substrate-2 in Hepatoma Cells

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