BIRC5对山羊睾丸细胞周期、凋亡的影响

doi:10.11843/j.issn.0366-6964.2023.04.015

Abstract

Abstract: The aim of the present study was to clone goat BIRC5, and to reveal the role of BIRC5 gene in regulating the cycle and apoptosis of goat testis cells through the overexpression or interference of BIRC5. These data will be beneficial for exploring the role of BIRC5 gene in goat. The BIRC5 gene sequence was cloned by RT-PCR from spleen tissue of 3 three-day-old healthy Jianzhou male goats with 3 kg of body weight, and sequenced for further bioinformatics analysis using online softwares. The clone recovery product was ligated to a eukaryotic expression vector to construct pcDNA3.1(+)-BIRC5. Effective siRNA was designed and screened based on goat BIRC5 gene sequence. After pcDNA3.1(+)-BIRC5 and siRNA were respectively transfected into goat testicular cells, the expression of BIRC5 protein was detected by Western blot, and cell cycle and apoptosis were detected by flow cytometry. Meanwhile, the expression of apoptosis-related genes Bax, Caspase3, Caspase7, Bcl-2, p53, BCL2L11 and PARP1 were detected by RT-qPCR. A length of 506 bp BIRC5 gene sequence was cloned successfully, including 28 bp of 5'UTR, 429 bp of CDS region, and 49 bp of 3'UTR,encoding 142 amino acids. The phylogenetic tree showed that BIRC5 had the closest relative to Bos taurus. The overexpression of BIRC5 gene inhibited the apoptosis of cells, and the cells were arrested in G2/M+S phase; it also down-regulated the expression of Caspase7, p53, BCL2L11, Bcl-2 and Bax gene mRNA, but did not significantly change the mRNA expression of Caspase3 and PARP1. siRNA interference of BIRC5 gene promoted the apoptosis of cells, and the cells were arrested in G0/G1 phase; it also up-regulated the expression of Caspase3, Caspase7 and PARP1 gene mRNA, and down-regulated the expression of p53 and Bcl-2 gene mRNA, but did not significantly change the mRNA expression of Bax and BCL2L11. Goat BIRC5 gene could inhibit the apoptosis of testis cells and promote the arrest of goat testicular cells in G2/M+S phase. These data may lay a foundation for further study of BIRC5 gene function.

Key words: goat, BIRC5 gene, overexpression, interference, cell cycle, cell apoptosis

CLC Number:

S827.3

DAN Yixin, YANG Lu, XIANG Hua, ZHANG Huanrong, REN Yupeng, YANG Falong, HE Honghong, ZHU Jiangjiang. Effects of BIRC5 on the Cycle and Apoptosis of Goat Testis Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1511-1524.

References 10

[1]	OH K, WILLETT W C, WU K N, et al.Calcium and vitamin D intakes in relation to risk of distal colorectal adenoma in women[J].Am J Epidemiol, 2007, 165(10):1178-1186.
[2]	刘俊.Survivin在不同年龄牦牛组织器官内表达检测及其多克隆抗体的制备[D].兰州:甘肃农业大学, 2019.LIU J.Expression of survivin in organs of different age yak and preparation of survivin polyclonal antibody[D].Lanzhou:Gansu Agricultural University, 2019.(in Chinese)
[3]	PAHLAVAN Y, KAHROBA H, SAMADI N, et al.Survivin modulatory role in autoimmune and autoinflammatory diseases[J].J Cell Physiol, 2019, 234(11):19440-19450.
[4]	ADAMS R R, CARMENA M, EARNSHAW W C.Chromosomal passengers and the (aurora) ABCs of mitosis[J].Trends Cell Biol, 2001, 11(2):49-54.
[5]	EBRAHIMIYAN H, ASLANI S, REZAEI N, et al.Survivin and autoimmunity; the ins and outs[J].Immunol Lett, 2018, 193:14-24.
[6]	SOHN H J, LEE J Y, LEE H J, et al.Simultaneous in vitro generation of CD8 and CD4 T cells specific to three universal tumor associated antigens of WT1, survivin and TERT and adoptive T cell transfer for the treatment of acute myeloid leukemia[J]. Oncotarget, 2017, 8(27):44059-44072.
[7]	ZHANG L, LIANG H, CAO W, et al.Downregulation of survivin by siRNA inhibits invasion and promotes apoptosis in neuroblastoma SH-SY5Y cells[J].Braz J Med Biol Res, 2014, 47(7):548-553.
[8]	SERPELONI J M, SPECIAN A F L, RIBEIRO D L, et al.Fridericia platyphylla (Cham.) L.G. Lohmann root extract exerts cytotoxic and antiproliferative effects on gastric tumor cells and downregulates BCL-XL, BIRC5, and MET genes[J].Hum Exp Toxicol, 2020, 39(3):338-354.
[9]	NARIMANI M, SHARIFI M, JALILI A.Knockout of BIRC5 gene by CRISPR/Cas9 induces apoptosis and inhibits cell proliferation in leukemic cell lines, HL60 and KG1[J].Blood Lymphat Cancer, 2019, 9:53-61.
[10]	YANG Y, LI D, LI Y L, et al.Low-temperature plasma suppresses proliferation and induces apoptosis in lung cancer cells by regulating the miR-203a/BIRC5 axis[J].Onco Targets Ther, 2020, 13:5145-5153.

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Effects of BIRC5 on the Cycle and Apoptosis of Goat Testis Cells

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