干扰lnc721对牛骨骼肌卫星细胞增殖与分化的影响

doi:10.16431/j.cnki.1671-7236.2022.09.003

Abstract

Abstract: 【Objective】 This experiment was aimed to explore the long non-coding RNA (lncRNA) that played a regulatory role in bovine skeletal muscle development, and study its effect on the expression of bovine skeletal muscle satellite cells in proliferation and differentiation, so as to provide reference for the study of bovine muscle development related mechanisms.【Method】 A lncRNA obtained by high-throughput sequencing in the previous test was named lnc721. The biological information of lnc721 was analyzed using NCBI database, the coding ability and subcellular localization of lnc721 were determined by CPC website and nucleo-cytoplasmic separation experiment, respectively.The designed and synthesized lnc721 siRNA was transfected into bovine skeletal muscle satellite cells, and the bovine skeletal muscle satellite cells were cultured and differentiated through a mature in vitro myoblast induced differentiation model.The expression changes of lnc721 in different stages of cell development were analyzed in detail by EdU experiment, Real-time quantitative PCR and Western blotting, respectively.【Result】 lnc721 was located on chromosome 18 of the whole bovine genome, and its protein coding capacity was-1.33129, which was mainly located in cytoplasm.After interference with lnc721 expression, the EdU positive cellular rates was extremely significantly increased, and the mRNA expression of cell proliferation marker genes Pax7 and Ki-67 were extremely significantly up-regulated (P<0.01).Western blotting results further proved that interference with lnc721 extremely significantly promoted the expression of Pax7 protein (P<0.01).The mRNA and protein expression of the cell differentiation marker MyHC gene were extremely significantly down-regulated after interfering with the expression of lnc721 at the cell differentiation stage (P<0.01).【Conclusion】 Interference with lnc721 could promote the proliferation of bovine skeletal muscle satellite cells and inhibit the process of myoblast differentiation.

Key words: lnc721; cattle; skeletal muscle satellite cells; proliferation; differentiation

CLC Number:

S823

TAN Haoyun, LIU Qian, HU Debao, ZHANG Linlin, LI Xin, DING Xiangbin, GUO Hong, GUO Yiwen. Effects of Interference lnc721 on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3292-3300.

References

[1] ZHANG Z K, LI J, GUAN D G, et al.A newly identified lncRNA MAR1 acts as a miR-487b sponge to promote skeletal muscle differentiation and regeneration[J].Journal of Cachexia Sarcopenia and Muscle, 2018, 9(3):613-626.
[2] YONG H, WU G M, CHEN J Y, et al.lncRNA MALAT1 accelerates skeletal muscle cell apoptosis and inflammatory response in sepsis by decreasing BRCA1 expression by recruiting EZH2[J].Molecular Therapy-Nucleic Acids, 2020, 3(6):97-108.
[3] LI J, YANG T T, TANG H F, et al.Inhibition of lncRNA MAAT controls multiple types of muscle atrophy by cis- and trans-regulatory actions[J].Molecular Therapy-Nucleic Acids, 2021, 29(3):1102-1119.
[4] ALEXANDER F P, EUGENE V K.Functional long non-coding RNAs evolve from junk transcripts[J].Cell, 2020, 9:47.
[5] 杨峰, 易凡, 曹慧青, 等.长链非编码RNA研究进展[J].遗传, 2014, 36(5):456-468. YANG F, YI F, CAO H Q, et al.The emerging landscape of long non-coding RNAs[J].Hereditas, 2014, 36(5):456-468.(in Chinese)
[6] 夏天, 肖丙秀, 郭俊明, 等.长链非编码RNA的作用机制及其研究方法[J].遗传, 2013, 35(3):269-280. XIA T, XIAO B X, GUO J M, et al.Acting mechanisms and research methods of long noncoding RNAs[J].Hereditas, 2013, 35(3):269-280.(in Chinese)
[7] CHOI S W, KIM H W, NAM J W.The small peptide world in long noncoding RNAs[J]. Brief Bioinform, 2019, 20(5):1853-1864.
[8] WU P, MO Y Z, PENG M, et al.Emerging role of tumor-related functional peptides encoded by lncRNA and circRNA[J].Molecular Cancer, 2020, 19(1):22.
[9] YU X H, ZHANG Y, LI T T, et al.Long non-coding RNA linc-RAM enhancesmyogenicdifferentiation by interacting with MyoD[J].Nature Communications, 2017, 8:14016.
[10] ZHU M, LIU J F, XIAO J, et al.lnc-mg is a long non-coding RNA that promotes myogenesis[J].Nature Communications, 2017, 8:14718.
[11] 杨洋, 许厚强, 陈伟, 等.从江香猪PDK4基因干扰载体的构建与检测[J].中国畜牧兽医, 2018, 45(4):873-880. YANG Y, XU H Q, CHEN W, et al.Construction and detection of the interference vector of PDK4 gene in Congjiang Xiang pig[J].China Animal Husbandry & Veterinary Medicine, 2018, 45(4):873-880.(in Chinese)
[12] CHEN M M, ZHANG L L, GUO Y W, et al.A novel lncRNA promotes myogenesis of bovine skeletal muscle satellite cells via PFN1-RhoA/Rac1[J].Journal of Cell and Molecular Medicine, 2021, 25(13):5988-6005.
[13] ZHANG X J, CHEN M M, LIU X F, et al.A novel lncRNA, lnc403, involved in bovine skeletal muscle myogenesis by medicating KRAS/Myf6[J].Gene, 2020, 751:144706.
[14] 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. WANG Y M, DAI Y, LIU X F, et al.Isolation, identification and differentiation of bovine skeletal muscle satellite cells[J].China Animal Husbandry & Veterinary Medicine, 2014, 41(7):142-147.(in Chinese)
[15] JIN C F, LI Y, DING X B, et al.lnc133b, a novel, long non-coding RNA, regulates bovine skeletal muscle satellits cell proliferation and differentiation by mediating miR-133b[J].Gene, 2017, 630:35-43.
[16] ZHANG W R, ZHANG H Z, WANG Y M, et al.miR-143 regulates proliferation and differentiation of bovine skeletal muscle satellite cells by targeting IGFBP5[J].In Vitro Cellular & Development Biology-Animal, 2016, 53(3):265-271.
[17] LENINI I, MORLANDO M, MANGIAVACCHI A, et al.A feed forward regulatory loop between the HuR protein and the long non coding RNA linc-MD1 controls early phases of muscle differentiation[J].Molecular Cell, 2013, 3:506-514.
[18] GONG C G, LI Z Z, RAMANUJAN K, et al.A long non-coding RNA, lncMyoD, regulates skeletal muscle differentiation by blocking IMP2-mediated mRNA translation[J].Developmental Cell, 2015, 34(2):181-191.
[19] 李艳, 陈明明, 张晓娟, 等.牛lncRNA-133a对骨骼肌卫星细胞增殖分化的影响[J].中国农业科学, 2019, 52(1):141-153. LI Y, CHEN M M, ZHANG X J, et al.Effects of bovine lncRNA-133a on the proliferation and differentiation of skeletal muscle satellite cells[J].Scientia Agricultura Sinica, 2019, 52(1):141-153.(in Chinese)
[20] GONG C G, LYNNE E, MAQU A Y.lncRNAs transactivate STAU1-mediated mRNA decay by duplexing with 3'UTRs via Alu elements[J].Nature, 2011, 470(7333):284-288.
[21] SONGC C, YANG Z X, JIANG R, et al.lncRNA IGF2 AS regulates bovine myogenesis through different pathways[J].Molecular Therapy-Nucleic Acids, 2020, 21:874-884.
[22] DU Z, SUN T, HACISULEYMAN E, et al.Integrative analyses reveal a long noncoding RNA-mediated sponge regulatory network inprostate cancer[J].Nature Communictions, 2016, 7:10982.
[23] MARCELLA C, DAVIDE C, IVANO L, et al.A long noncoding RNA controls muscle differentiation by functioning as a competing endogenous RNA[J].Cell, 2011, 147(2):358-369.
[24] LV W, ZHAO S Y, HOU Y Q, et al.Single nucleotide polymorphisms of porcine lncMGPF regulate meat production traits by affecting RNA stability[J].Frontiers in Cell and Developmental Biology, 2021, 9:731712.
[25] ZHAI L L, WAN X, WU R M, et al.linc-RAM promotes muscle cell differentiation via regulating glycogen phosphorylase activity[J].Cell Regeneration, 2022, 11(1):8.
[26] TAN Y, GAN M L, SHEN L Y, et al.Profiling and functional analysis of long noncoding RNAs and mRNAs during porcine skeletal muscle development[J]. International Journal of Molecular Sciences, 2021, 22(2):503.
[27] 周瑞, 王以鑫, 龙科任, 等.lncRNA调控骨骼肌发育的分子机制及其在家养动物中的研究进展[J].遗传, 2018, 40(4):292-304. ZHOU R, WANG Y X, LONG K R, et al.Molecular mechanism of lncRNA regulating skeletal muscle development and its research progress in domestic animals[J].Hereditas, 2018, 40(4):292-304.(in Chinese)
[28] SCHONK D M, KUIJPERS H J, DRUNEN E V, et al.Assignment of the gene(s) involved in the expression of the proliferation-related Ki-67 antigen to human chromosome 10[J].Human Cenetics, 1989, 83(3):297-299.
[29] BUCKINGHAM M, RELAIX F.PAX3 and PAX7 as upstream regulators of myogenesis[J].Seminars in Cell & Developmental Biology, 2015, 44:115-125.
[30] TAKAGAKI Y, YAMAGISHI H, MATSUOKA R.Factors involved in signal transduction during vertebrate myogenesis[J].International Review of Cell and Molecular Biology, 2012, 296:187-272.
[31] SCHIAFFINO S, REGGIANI C.Myosin isoforms in mammalian skeletal muscle[J].Journal of Applied Physiology, 1994, 77(2):493-501.
[32] 张俊星, 朱菲菲, 李燕, 等.干扰lnc4351对牛骨骼肌卫星细胞增殖与成肌分化的影响[J].中国畜牧兽医, 2019, 46(3):747-755. ZHANG J X, ZHU F F, LI Y, et al.Effect of lnc4351 interference on the proliferation and differentiation of bovine skeletal muscle satellite cells[J].China Animal Husbandry & Veterinary Medicine, 2019, 46(3):747-755.(in Chinese)

Effects of Interference lnc721 on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	XIAO Peng, SHANG Jianghua, YANG Chunyan, LI Mengqi, DUAN Anqin, MA Xiaoya, FENG Chao, HUANG Chenqian, ZHANG Bo, ZHOU Jinchen, WEI Kelong, ZHENG Wei, ZHENG Haiying. Effects of Alpha-linolenic Acid on Buffalo Ovarian Granulosa Cells Cultured in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2380-2387.
[2]	LI Can, LI Chuchu, ZENG Wei, ZHANG Ning, JI Chunxiao, CHEN Tao. Recombinant Expression and Function Study of RegⅢγ Protein in Porcine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2414-2426.
[3]	ZHANG Dianqi, MA Xinhao, YANG Zhimei, MEI Chugang, ZAN Linsen. Bioinformatics Analysis of PAI1 Gene and Its Effect on the Proliferation and Differentiation of Intramuscular Adipocytes in Qinchuan Beef Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1729-1741.
[4]	PENG Yangxiang, YANG Zhenbiao, YAN Kuiyou, WANG Han, HU De, WANG Zun, LIU Ning, ZHAO Liansheng. From Artificial to Intelligent:Research Progress of Individual Idendification Technology for Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1855-1866.
[5]	MENG Chaoqun, LI Chengping, ZHAO Wei, QIN Xuyong, ZHOU Guoli. Effect of miR-137-3p on Adipogenic Differentiation of Preadipocytes 3T3-L1 by Targeting MAX Gene [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1928-1937.
[6]	LI Zhongbo, HOU Qianghong, DUAN Deyong. Haplotypes Diversity,Genetic Differentiation and Phylogenetic Analysis of Aedes albopictus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1452-1460.
[7]	ZHEN Zhen, WANG Mei, WANG Yimin, HU Debao, ZHANG Linlin, LI Xin, GUO Yiwen, GUO Hong, DING Xiangbin. Effect of RNA Methylation Transfer Enzyme METTL3 on the Proliferation and Myogenic Differentiation of Bovine Skeletal Muscle Satellite Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1025-1036.
[8]	LI Mingbo, GAO Qingshan, LI Guanhao, ZHANG Kui, CUI Lianhua, HOU Lina. Effects of Chinese Herbal Medicine Feed Additives on Meat Quality and Flavor Substances in Yanbian Yellow Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 531-542.
[9]	TIAN Shuyue, SHEN Yingchao, WANG Xisheng, WANG Min, YI Minna, Tseweendolmaa, TAO Li, ZHAO Bilig, Manglai, Gerelchimeg. Study on Establishment of Immortalized Cell System of Equine Fetal Fibroblasts in Mongolian Horse [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 46-57.
[10]	SUN Haobin, XUE Yuyang, NIU Yujie, NIU Junli, ZHANG Wenju, CHEN Cheng. Effects of Whole Triticale Hay on Nutrient Apparent Digestibility,Serum Biochemical Indices and Rumen Fermentation of Breeding Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 106-114.
[11]	GAO Yefan, SONG Hanan, WANG Yunan, WU Yue, NIU Ruili, ZONG Xianchun, GUAN Weijun. Study on the Isolation, Culture and Differentiation Potential of Limbal Stem Cells from Beijing You Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3372-3381.
[12]	XU Haoqi, XU Jingyi, HU Lirong, ZHANG Fan, LUO Hanpeng, ZHANG Hailiang, SHI Rui, LI Xiang, LIU Lin, LIU Qiaoxiang, GUO Gang, WANG Yachun. Polymorphism of PIK3CB Gene and Its Association Analysis with Reproductive and Milk Production Traits in Chinese Holstein Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3438-3452.
[13]	WANG Xuefang, GUI Rong, CHEN Gongyi, WANG Wei, ZHANG Hongying, WANG Xuebing, LI Xiao. Clinical Study of Lianhuachaiqin Soluble Powder on Chicken Syndrome of Wind-heat Invading Lung [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3610-3621.
[14]	WANG Nan, FENG Baoliang, ZHENG Yunxi, HUANG Lei, WANG Yue, XU Songsong, ZHANG Xiuling, LIU Zhiguo, LI Kui, MU Yulian. Effects of WIP1 Gene on Proliferation and Differentiation of 3T3-L1 Preadipocytes and Its Expression in Different Growth Stages of Mice [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2869-2879.
[15]	ZHANG Xueping, SHI Bingang, JIN Xiayang, WANG Xiangyan, LAN Lijuan, SHI Yu, QI Youpeng, ZHAO Shijie, LI Shaobin, HU Jiang. Expression Characterization Analysis of KLF7 Gene and Effects of Its Overexpression on Proliferation and Differentiation of Preadipocytes in Tibetan Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2920-2930.