NR2F2基因调控猪PK15细胞增殖和凋亡的研究

doi:10.11843/j.issn.0366-6964.2023.08.011

Abstract

Abstract: This study aimed to explore the effect of NR2F2 gene on the proliferation and apoptosis of porcine PK15 cells. In this study, the effect of overexpression of NR2F2 gene on cell proliferation was detected by qRT-PCR, Western blot, CCK-8, EdU, flow cytometry, scratch assay and other methods in porcine PK15 cells. CO-IP technology was used to verify the binding ability of NR2F2 to Smad4 protein. The effect of NR2F2 gene on Smad4 gene expression and the expression of downstream gene Cyclin B, CDK1, CDK4, Cyclin D1 were detected by qRT-PCR. The expression of downstream genes and proliferating genes Ki67, PCNA were detected by rescue experiment overexpressed NR2F2 gene and interfered with Smad4 gene. The results showed that overexpression of NR2F2 gene significantly upregulated the expression of Ki67(P<0.01), and significantly upregulated the expression of PCNA(P<0.05), promoted cell cycle progression, inhibited cell apoptosis, and promoted cell proliferation. The CO-IP results proved that NR2F2 and Smad4 proteins were physically bound. After overexpression of NR2F2 gene, the expression of Smad4 gene increased significantly (P<0.01), and the expression of downstream regulated genes by Smad4 gene Cyclin B, CDK1, CDK4 and Cyclin D1 significantly increased (P<0.01). After interfering with NR2F2 gene, the expression of Smad4 gene significantly decreased (P<0.01), and the expression of downstream genes regulated by Smad4 gene was significantly decreased (P<0.01). The rescue experiments showed that after overexpression of NR2F2 gene, interfering with the expression of Smad4 gene could significantly reduce the expression of Cyclin D1, Cyclin B, CDK1, Ki67 and PCNA (P<0.01,P<0.05). In summary, overexpression of NR2F2 gene promoted cell proliferation and inhibited apoptosis, and NR2F2 protein could bind to Smad4 protein and affect the expression of Smad4, which in turn affected the expression of downstream genes.

Key words: NR2F2 gene, Smad4 gene, cell proliferation, PK15 cell

CLC Number:

S828.2

ZHANG Wanfeng, ZHAO Tianzhi, LI Jiao, YOU Ziwei, YANG Yang, CAI Chunbo, GAO Pengfei, CAO Guoqing, GUO Xiaohong, LI Bugao. Study on NR2F2 Gene Regulating Proliferation and Apoptosis of Porcine PK15 Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3242-3251.

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Study on NR2F2 Gene Regulating Proliferation and Apoptosis of Porcine PK15 Cells

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