X Zuo1, G Li, T Luo, J Li, Y Liu, M Luo. 1. Affiliated Ruijin Hospital of Shanghai Second Medical University, Shanghai Institute of Endocrinology, Shanghai 200025, China. xiangshengzuo@yahoo.com
Abstract
OBJECTIVE: To express mouse peroxisome proliferator activated receptor gamma 2 (mPPAR gamma 2) in NIH3T3 fibroblasts mediated by the recombinant retrovirus and study its function. METHODS: The mPPAR gamma 2 gene was subcloned into retrovirus vector pGCEN to generate the recombinant pGCEN/mPPAR gamma 2. Then it was packaged into PA317 cells and selected with G418. Viral supernatants were harvested and then used to infect NIH3T3 fibroblasts. PPAR gamma activator 5, 8, 11, 14-eicosatetraynoic acid (ETYA) was used to induce the mPPAR gamma 2-expressing NIH3T3 cells into adipocyte differentiation. RESULTS: The recombinant retrovirus pGCEN/mPPAR gamma 2 was constructed, and the higher titers of the viral supernatants were obtained. mPPAR gamma 2 was expressed in NIH3T3 cells mediated by the recombinant retrovirus. Lipid accumulation obviously existed in these induced adipocytes which morphologically resembled mature adipocytes in vivo and expressed tissue specific adipocyte P2 (AP2) and Leptin genes. CONCLUSIONS: An adipocyte differentiation model in vitro was successfully established. The work is the basis for further research on the molecular mechanism of adipocyte differentiation induced by PPAR gamma 2.
OBJECTIVE: To express mouseperoxisome proliferator activated receptor gamma 2 (mPPAR gamma 2) in NIH3T3 fibroblasts mediated by the recombinant retrovirus and study its function. METHODS: The mPPAR gamma 2 gene was subcloned into retrovirus vector pGCEN to generate the recombinant pGCEN/mPPAR gamma 2. Then it was packaged into PA317 cells and selected with G418. Viral supernatants were harvested and then used to infect NIH3T3 fibroblasts. PPAR gamma activator 5, 8, 11, 14-eicosatetraynoic acid (ETYA) was used to induce the mPPAR gamma 2-expressing NIH3T3 cells into adipocyte differentiation. RESULTS: The recombinant retrovirus pGCEN/mPPAR gamma 2 was constructed, and the higher titers of the viral supernatants were obtained. mPPAR gamma 2 was expressed in NIH3T3 cells mediated by the recombinant retrovirus. Lipid accumulation obviously existed in these induced adipocytes which morphologically resembled mature adipocytes in vivo and expressed tissue specific adipocyte P2 (AP2) and Leptin genes. CONCLUSIONS: An adipocyte differentiation model in vitro was successfully established. The work is the basis for further research on the molecular mechanism of adipocyte differentiation induced by PPAR gamma 2.