腺病毒载体介导Smad3基因在秦川牛前体脂肪细胞中的过表达和沉默

doi:10.11843/j.issn.0366-6964.2018.09.005

Abstract

Abstract:

The purpose of this study was to investigate the biological function of Smad3 gene during Qinchuan cattle preadipocytes differentiation, and provide a theoretical basis for clarifying the role of TGF-β signaling pathway in the growth and development of cattle adipose tissue by overexpressing and interfering Smad3 gene in Qinchuan cattle preadipocytes. In this study, overexpression adenovirus pDC316-EGFP-bSMAD3 and interference adenovirus pDC316-EGFP-ShRNA-bSMAD3-1405 were successfully obtained by using AdMaxTM recombinant adenovirus packaging system reconstructed with Ad5 adenovirus. Then we detected the mRNA expression of Smad3 gene and adipogenesis-related genes by real-time fluorescence quantitative PCR after the adenovirus infected bovine preadipocytes. The results showed that, compared to the control group, the expression of Smad3 gene in Qinchuan cattle preadipocytes were significantly increased about 1 031, 1 286, 633 times, respectively(P<0.001) when bovine preadipocytes were infected by pDC316-EGFP-bSMAD3 for 3, 6, 9 d. Meanwhile, the interference group's (pDC316-EGFP-ShRNA-bSMAD3-1405) expression of Smad3 gene in cattle preadipocytes reduced by 70%, 55% and 57%, respectively (P<0.01) compared with the control group. Moreover, both Oil Red O staining assay and qRT-PCR results demonstrated that Smad3 gene suppressed the lipid accumulation in preadipocytes and markedly down-regulated the expression of adipogenesis-related gene PPARγ, FABP4, C/EBPα and C/EBPβ. In conclusion, Smad3 gene is successfully overexpressed and interfered in Qinchuan cattle preadipocytes mediated by adenovirus, and plays an important role in adipogenic differentiation process in Qinchuan cattle preadipocytes.

Key words: Qinchuan cattle, Smad3 gene, preadipocyte, adenovirus vector

CLC Number:

S823.2

ZHANG Le, NING Yue, LI Pei-wei, WANG Hong-bao, ZAN Lin-sen. The Overexpression and Interference of Smad3 Gene in Qinchuan Cattle Preadipocytes Mediated by Adenovirus Vector[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 1840-1850.

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The Overexpression and Interference of Smad3 Gene in Qinchuan Cattle Preadipocytes Mediated by Adenovirus Vector

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