Differentiation of NIH3T3 fibroblasts into adipocytes induced by peroxisome proliferator activated receptor gamma 2 expression.
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.
Zuo
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X
Affiliated Ruijin Hospital of Shanghai Second Medical University, Shanghai Institute of Endocrinology, Shanghai 200025, China. xiangshengzuo@yahoo.com
Li
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Luo
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Li
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Liu
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Luo
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D016475:3T3 Cells; D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D018076:DNA, Complementary / Q000235:genetics; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D020738:Leptin / Q000235:genetics; D051379:Mice; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D012313:RNA / Q000235:genetics / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000502:physiology; D012190:Retroviridae / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*; D014162:Transfection
D002352:Carrier Proteins; D018076:DNA, Complementary; C082667:Fabp5 protein, mouse; C086936:Fabp7 protein, mouse; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D020738:Leptin; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D012313:RNA