UCH-L3通过影响RAD51调控牛骨骼肌卫星细胞增殖与成肌分化

doi:10.19640/j.cnki.jtau.2023.02.001

Journal of Tianjin Agricultural University ›› 2023, Vol. 30 ›› Issue (2): 1-6.doi: 10.19640/j.cnki.jtau.2023.02.001

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UCH-L3 regulating the proliferation and differentiation of bovine skeletal muscle satellite cells by influencing RAD51

Hu Lei, Wang Yimin, Zhang Linlin, Li Xin, Guo Yiwen, Ding Xiangbin^{Corresponding Author}

Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry, College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300392, China

Received:2021-08-23 Online:2023-04-30 Published:2023-06-28

Abstract

Abstract: The purpose of this study was to explore the mechanism of carboxyl terminal hydrolase L3（UCH-L3）in ubiquitin proteasome system regulating the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells by affecting RAD51. Using the established in vitro myogenic differentiation model of bovine skeletal muscle satellite cells, this study simulated the development process of skeletal muscle in vitro, detected whether uch-l3 interacts with RAD51 by immunoprecipitation technology, and whether uch-l3 regulates the ubiquitination level of RAD51, designed and synthesized si-RNA of RAD51, transfected bovine skeletal muscle satellite cells, and screened si-RNA with significant interference effect, detected the changes of the expression levels of proliferation and differentiation marker genes Pax7 and MyHC after interfering with Rad51, and comprehensively analyzed the mechanism of UCH-L3 regulating the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells by affecting Rad51. The results showed that the protein levels of RAD51 increased significantly after over-expression of UCH-L3, and UCH-L3 could regulate the expression level of RAD51. UCH-L3 interacts with RAD51 in vivo. UCH-L3 can affect the protein stability of RAD51. Interfering with UCH-L3 expression can shorten the half-life of RAD51. UCH-L3 can regulate the ubiquitination level of RAD51. After interfering with RAD51, the protein level of Pax7, a marker of bovine skeletal muscle satellite cell proliferation, was not different from that of the control group, and the protein level of MyHC, a marker of differentiation, was significantly lower than that of the control group（P<0.05）. Interfering with the expression of RAD51 had no significant effect on cell proliferation, but could inhibit the myogenic differentiation of bovine skeletal muscle satellite cells. The results showed that UCH-L3 could regulate the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells by affecting RAD51.

Key words: RAD51, UCH-L3, bovine skeletal muscle satellite cells, proliferation, myogenic differentiation

CLC Number:

Hu Lei, Wang Yimin, Zhang Linlin, Li Xin, Guo Yiwen, Ding Xiangbin. UCH-L3 regulating the proliferation and differentiation of bovine skeletal muscle satellite cells by influencing RAD51[J]. Journal of Tianjin Agricultural University, 2023, 30(2): 1-6.

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References

[1] NANDI D,TAHILIANI P,KUMAR A,et al.The ubiquitin-proteasome system[J]. Journal of Biosciences,2006,31(1):137-155.
[2] 胡蕾,朱菲菲,王轶敏,等. 干扰UCH-L3对牛骨骼肌卫星细胞增殖与分化的影响[J]. 中国畜牧杂志,2020,56(11):100-104.
[3] SHINOHARA A,OGAWA H,MATSUDA Y,et al.Cloning of human,mouse and fission yeast recombination genes homologous to RAD51 and recA[J]. Nature Genetics,1993,4(3):239-243.
[4] TAYLOR M R G,ŠPÍREK M,CHAURASIYA K R,et al. Rad51 paralogs remodel pre-synaptic Rad51 filaments to stimulate homologous recombination[J]. Cell,2015,162(2):271-286.
[5] TCHOU J,KASAI H,SHIBUTANI S,et al.8-oxoguanine (8-hydroxyguanine) DNA glycosylase and its substrate specificity[J]. Proceedings of the National Academy of Sciences of the United States of America,1991,88(11):4690-4694.
[6] CHEVILLARD S,RADICELLA J P,LEVALOIS C,et al.Mutations in OGG1,a gene involved in the repair of oxidative DNA damage,are found in human lung and kidney tumours[J]. Oncogene,1998,16(23):3083-3086.
[7] WODITSCHKA S,EVANS L,DUCHNOWSKA R,et al.DNA double-strand break repair genes and oxidative damage in brain metastasis of breast cancer[J]. Journal of the National Cancer Institute,2014,106(7):766-776.
[8] WIEGMANS A P,AL-EJEH F,CHEE N,et al.Rad51 supports triple negative breast cancer metastasis[J]. Oncotarget,2014,5(10):3261-3272.
[9] KO J C,CHEN J C,WANG T J,et al.Astaxanthin down-regulates Rad51 expression via inactivation of AKT kinase to enhance mitomycin C-induced cytotoxicity in human non-small cell lung cancer cells[J]. Biochemical Pharmacology,2016,105:91-100.
[10] LUO K,LI L,LI Y,et al.A phosphorylation- deubiquitination cascade regulates the BRCA2-RAD51 axis in homologous recombination[J]. Genes & Development,2016,30(23):2581-2595.
[11] 王轶敏,代阳,刘新峰,等. 牛骨骼肌卫星细胞的分离鉴定和诱导分化[J]. 中国畜牧兽医,2014,41(7):142-147.
[12] HOU D,XU G,ZHANG C,et al.Berberine induces oxidative DNA damage and impairs homologous recombination repair in ovarian cancer cells to confer increased sensitivity to PARP inhibition[J]. Cell Death & Disease,2017,8(10):e3070.
[13] WANG B,HOU D,LIU Q,et al.Artesunate sensitizes ovarian cancer cells to cisplatin by downregulating RAD51[J]. Cancer Biology & Therapy,2015,16(10):1548-1556.
[14] GASPARINI P,LOVAT F,FASSAN M,et al.Protective role of miR-155 in breast cancer through RAD51 targeting impairs homologous recombination after irradiation[J]. Proceedings of the National Academy of Sciences of the United States of America,2014,111(12):4536-4541.
[15] CHEN Q,CAI D,LI M,et al.The homologous recombination protein RAD51 is a promising therapeutic target for cervical carcinoma[J]. Oncology Reports,2017,38(2):767-774.

UCH-L3 regulating the proliferation and differentiation of bovine skeletal muscle satellite cells by influencing RAD51

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