Research Article

Construction of suppressor of cytokine signaling 2 (SOCS2) adenoviral overexpression vector and its impact on growth-hormone-induced lipolysis in swine primary adipocytes

Published: April 17, 2013
Genet. Mol. Res. 12 (2) : 1283-1293 DOI: 10.4238/2013.January.9.2

Abstract

We investigated the effect of overexpression suppressor of cytokine signaling 2 (SOCS2) on lipolysis in swine primary adipocytes (pAd) induced by growth hormone (GH). We constructed pAd-SOCS2 adenoviral overexpression vectors to infect HEK293 cells for virus packaging and propagation. Cultured swine primary adipocytes were infected with virus particles; after 48 h the infected adipocytes were treated with 500 ng GH/mL in the growth medium. Lipometabolism-related gene expressions were detected at 0, 0.25, 0.5, 1, 2, and 4 h, by measuring mRNA and protein levels. The pAd-SOCS2 overexpression vector was successfully constructed and the concentration of titrated virus was 1.2 x 109 PFU/mL. We found that virus infection significantly increased SOCS2 mRNA and protein levels in swine primary adipocytes. Overexpression of SOCS2 significantly inhibited the increase in fatty acid synthase, adipose triglyceride lipase mRNA, and protein expression at 0.5 h. However, after 0.5 h, this inhibition was not significant. We concluded that overexpression of SOCS2 inhibited the increase in lipolysis induced by GH in swine primary adipocytes; this could provide a basis for studies of lipometabolism.

We investigated the effect of overexpression suppressor of cytokine signaling 2 (SOCS2) on lipolysis in swine primary adipocytes (pAd) induced by growth hormone (GH). We constructed pAd-SOCS2 adenoviral overexpression vectors to infect HEK293 cells for virus packaging and propagation. Cultured swine primary adipocytes were infected with virus particles; after 48 h the infected adipocytes were treated with 500 ng GH/mL in the growth medium. Lipometabolism-related gene expressions were detected at 0, 0.25, 0.5, 1, 2, and 4 h, by measuring mRNA and protein levels. The pAd-SOCS2 overexpression vector was successfully constructed and the concentration of titrated virus was 1.2 x 109 PFU/mL. We found that virus infection significantly increased SOCS2 mRNA and protein levels in swine primary adipocytes. Overexpression of SOCS2 significantly inhibited the increase in fatty acid synthase, adipose triglyceride lipase mRNA, and protein expression at 0.5 h. However, after 0.5 h, this inhibition was not significant. We concluded that overexpression of SOCS2 inhibited the increase in lipolysis induced by GH in swine primary adipocytes; this could provide a basis for studies of lipometabolism.