干扰SERBP1表达对山羊鼻甲骨原代细胞增殖、周期及凋亡的作用

doi:10.11843/j.issn.0366-6964.2022.08.028

Abstract

Abstract: The aim of this study was to clone goat SERBP1, and to reveal the role of interfering with SERBP1 gene on the proliferation, cell cycle and apoptosis of goat turbinate bone primary cells. These data may provide important experimental data for further mechanism study underlying cell apoptosis induced by Orf virus (ORFV). With the Jianzhou goats as the experimental animal, the SERBP1 gene sequence was cloned by RT-PCR and analyzed by bioinformatics. Real-time fluorescent quantitative PCR(RT-qPCR) was performed to determine the expression of SERBP1 in different tissues, and screening the most effective interference sequence. The MTT method was used to detect the effects of siRNA interference SERBP1 on cell proliferation of goat turbinate bone cells, and the effects of siRNA interference with SERBP1 on the cell cycle and apoptosis were detected by flow cytometry, and the effects on apoptosis-related genes P53, Caspase7 and Caspase3 were detected by RT-qPCR. A total length of 1 293 bp SERBP1 gene sequence was cloned successfully, including 44 bp of 5'UTR, 1 179 bp of CDS region, and 70 bp of 3'UTR,encoding 392 amino acids, with no signal peptide and mainly located in the nucleus. It was predicted that SERBP1 might interact with 10 proteins including RPL31 and RPS3. The expression mRNA of SERBP1 gene was the highest in goat pancreas, and an effective interference sequence with an interference efficiency of 70% was selected. siRNA interference of SERBP1 gene inhibited the proliferation and apoptosis of goat turbinate primary cells, and the cells were arrested in G₀/G₁ phase. The mRNA expression of Caspase3, Caspase7, BCL2L11 and Bax gene was down-regulated but no significant difference was determined in the mRNA expression of Bcl-2, P53, and PARP1 gene. Interference with goat SERBP1 gene could inhibit the proliferation and apoptosis of goat turbinate bone cells and induce cell cycle arrest. These data may lay a foundation for further study of SERBP1 gene function, and also facilitate the mechanism research underlying cell apoptosis inhibited by ORFV125 gene.

Key words: goat, SERBP1 gene, interference, cell cycle, cell apoptosis

CLC Number:

S826.2

XU Songwei, XIANG Hua, ZHANG Huanrong, YANG Lu, WANG Xianjun, SALANG Wenzhu, ZHU Jiangjiang. Effects of Interfering SERBP1 on the Cell Proliferation, Cell Cycle and Apoptosis in Goat Turbinate Bone Primary Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2708-2720.

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Effects of Interfering SERBP1 on the Cell Proliferation, Cell Cycle and Apoptosis in Goat Turbinate Bone Primary Cells

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