干扰和过表达CHRNG对牛成肌细胞增殖分化的影响

doi:10.11843/j.issn.0366-6964.2024.10.011

摘要/Abstract

摘要：

旨在研究CHRNG基因对牛成肌细胞增殖分化的影响及其潜在的分子作用途径。本研究从健康3日龄秦川牛背最长肌和后腿肌中分离到成肌细胞，利用腺病毒在秦川牛成肌细胞中过表达及干扰CHRNG基因，分为干扰组(sh-CHRNG)、干扰对照组(sh-NC)、过表达组(OE-CHRNG)、过表达对照组(OE-NC)，每组3个重复，采用CCK-8、EdU、qRT-PCR、Western blot、免疫荧光染色等方法分别检测了干扰及过表达CHRNG基因对牛成肌细胞增殖分化的作用；通过RNA-Seq进一步筛选差异基因，富集信号通路。结果表明，过表达及干扰CHRNG均显著下调了细胞周期因子PCNA、CCNB1、CCND2的表达(P < 0.01)，显著上调了CDKN1A的表达(P < 0.01)，减少了发生增殖的细胞数量(CCK-8，P < 0.01)且减少了处于S复制期的阳性细胞比例(EdU)。在牛成肌细胞上过表达和干扰CHRNG并诱导分化后D2、4、6进行形态学观察和免疫荧光染色，结果表明干扰和过表达CHRNG抑制牛成肌细胞的分化和肌管形成。qRT-PCR和Western blot结果表明，过表达及干扰CHRNG下调了MYOD1、MYOG、MYH3基因mRNA和蛋白水平的表达(P < 0.01)。通过RNA-Seq测序分析发现，过表达CHRNG筛选到的差异基因主要富集在内质网蛋白质加工、IL-17、甲状腺激素合成、PPAR、PI3K-Akt等信号通路，KEGG分析富集的前20条通路中有10条通路与细胞分化相关；干扰CHRNG筛选到的差异基因主要富集在轴突导向、MAPK、PI3K-Akt等信号通路，KEGG分析富集的前20条通路中有8条通路与细胞分化相关。本研究结果表明，过表达和干扰CHRNG基因均能抑制牛成肌细胞的增殖和分化，且过表达和干扰CHRNG引起的差异基因所富集的通路大多与细胞分化相关。

关键词: CHRNG基因, 增殖和分化, 成肌细胞, RNA-Seq

Abstract:

This study aimed to investigate the effect of CHRNG gene on the proliferation and differentiation of bovine myoblasts and its potential molecular pathways. In this study, adenovirus was used to overexpress and interfere with CHRNG gene in Qinchuan cattle myoblasts. This study isolated myoblasts from the longissimus dorsi and hind leg muscles of healthy 3-day-old Qinchuan cattle. Adenovirus overexpression and interference with CHRNG gene were used in Qinchuan cattle myoblasts, which were divided into interference group (sh-CHRNG), interference control group (sh-NC), overexpression group (OE-CHRNG), and overexpression control group (OE-NC), with 3 replicates in each group. CCK-8, EdU, qRT-PCR, Western blot, immunofluorescence staining and other methods were used to detect the effects of interference and overexpression of CHRNG gene on the proliferation and differentiation of bovine myoblasts; Further screening of differential genes and enrichment of signaling pathways were performed through RNA-Seq. The results showed that overexpression and interference with CHRNG significantly downregulated the expression of cell cycle factors PCNA, CCNB1, and CCND2 (P < 0.01), significantly upregulated the expression of CDKN1A (P < 0.01), reduced the number of proliferating cells (CCK-8, P < 0.01), and reduced the proportion of positive cells in the S replication phase (EdU). Overexpression and interference of CHRNG on bovine myoblasts and induction of differentiation were observed by morphological observation and immunofluorescence staining on D2, D4, and D6. The results showed that interference and overexpression of CHRNG inhibited the differentiation of bovine myoblasts and myotube formation. The qRT-PCR and Western blot results showed that overexpression and interference with CHRNG downregulated the mRNA and protein levels of MYOD1, MYOG and MYH3 genes (P < 0.01). Through RNA-Seq sequencing analysis, it was found that the differentially expressed genes screened by overexpressing CHRNG were mainly enriched in endoplasmic reticulum protein processing, IL-17, thyroid hormone synthesis, PPAR, PI3K-Akt and other signaling pathways. Among the top 20 enriched pathways analyzed by KEGG, 10 pathways were related to cell differentiation; The differentially expressed genes screened by CHRNG were mainly enriched in signaling pathways such as axon guidance, MAPK, and PI3K-Akt. Among the top 20 enriched pathways analyzed by KEGG, 8 pathways were related to cell differentiation. The results suggested that overexpression and interference with CHRNG gene can inhibit the proliferation and differentiation of bovine myoblasts, and the pathways enriched by differential genes caused by overexpression and interference with CHRNG are mostly related to cell differentiation.

Key words: CHRNG gene, proliferation and differentiation, myoblast, RNA-Seq

中图分类号:

S823.2

毛晓宇, 杜嘉伟, 汤嘉玉, 潘金海, 蒋蕾, 孙小磊, 昝林森, 王洪宝. 干扰和过表达CHRNG对牛成肌细胞增殖分化的影响[J]. 畜牧兽医学报, 2024, 55(10): 4360-4376.

Xiaoyu MAO, Jiawei DU, Jiayu TANG, Jinhai PAN, Lei JIANG, Xiaolei SUN, Linsen ZAN, Hongbao WANG. Effects of CHRNG Gene on Proliferation and Differentiation of Bovine Myoblasts and Its Mechanism[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4360-4376.

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参考文献 45

1	SHAHRIYARI M , ISLAM M R , SAKIB S M , et al. Engineered skeletal muscle recapitulates human muscle development, regeneration and dystrophy[J]. J Cachexia Sarcopenia Muscle, 2022, 13 (6): 3106- 3121. doi: 10.1002/jcsm.13094
2	CHOI K H , YOON J W , KIM M , et al. Muscle stem cell isolation and in vitro culture for meat production: a methodological review[J]. Compr Rev Food Sci Food Saf, 2021, 20 (1): 429- 457. doi: 10.1111/1541-4337.12661
3	SUN Y J , LIU K P , HUANG Y Z , et al. Differential expression of FOXO1 during development and myoblast differentiation of Qinchuan cattle and its association analysis with growth traits[J]. Sci China Life Sci, 2018, 61 (7): 826- 835. doi: 10.1007/s11427-017-9205-1
4	YAN E F , GUO J X , YIN J D . Nutritional regulation of skeletal muscle energy metabolism, lipid accumulation and meat quality in pigs[J]. Anim Nutr, 2023, 14, 185- 192. doi: 10.1016/j.aninu.2023.04.009
5	YU M B , FENG Y Q , YAN J M , et al. Transcriptomic regulatory analysis of skeletal muscle development in landrace pigs[J]. Gene, 2024, 915, 148407. doi: 10.1016/j.gene.2024.148407
6	刘媛, 李溪月, 张维娅. MMP14调控骨骼肌卫星细胞分化的分子机制研究[J]. 畜牧兽医学报, 2024, 55 (4): 1592- 1604.
	LIU Y , LI X Y , ZHANG W Y . Molecular mechanism of MMP14 regulating skeletal muscle satellite cell differentiation[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55 (4): 1592- 1604.
7	CAI S F , WANG X Y , XU R , et al. KLF4 regulates skeletal muscle development and regeneration by directly targeting P57 and Myomixer[J]. Cell Death Dis, 2023, 14 (9): 612. doi: 10.1038/s41419-023-06136-w
8	RAI M , KATTI P , NONGTHOMBA U . Spatio-temporal coordination of cell cycle exit, fusion and differentiation of adult muscle precursors by Drosophila erect wing (Ewg)[J]. Mech Dev, 2016, 141, 109- 118. doi: 10.1016/j.mod.2016.03.004
9	TERRUZZI I , VACANTE F , SENESI P , et al. Effect of hazelnut oil on muscle cell signalling and differentiation[J]. J Oleo Sci, 2018, 67 (10): 1315- 1326. doi: 10.5650/jos.ess18086
10	QUINN M E , GOH Q , KUROSAKA M , et al. Myomerger induces fusion of non-fusogenic cells and is required for skeletal muscle development[J]. Nat Commun, 2017, 8, 15665. doi: 10.1038/ncomms15665
11	ABMAYR S M , PAVLATH G K . Myoblast fusion: lessons from flies and mice[J]. Development, 2012, 139 (4): 641- 656. doi: 10.1242/dev.068353
12	PANG K T , LOO L S W , CHIA S , et al. Insight into muscle stem cell regeneration and mechanobiology[J]. Stem Cell Res Ther, 2023, 14 (1): 129. doi: 10.1186/s13287-023-03363-y
13	WANG Y J , LU J Q , LIU Y J . Skeletal muscle regeneration in cardiotoxin-induced muscle injury models[J]. Int J Mol Sci, 2022, 23 (21): 13380. doi: 10.3390/ijms232113380
14	BELGRANO A , RAKICEVIC L , MITTEMPERGHER L , et al. Multi-tasking role of the mechanosensing protein Ankrd2 in the signaling network of striated muscle[J]. PLoS One, 2011, 6 (10): e25519. doi: 10.1371/journal.pone.0025519
15	SANDWEISS A J , PATEL S , BADER M Y , et al. A truncating variant of CHRNG as a cause of Escobar syndrome: a multiple pterygium syndrome subtype[J]. J Pediatr Genet, 2022, 11 (2): 144- 146. doi: 10.1055/s-0040-1715640
16	KAPUR A , DAVIES M , DRYDEN W F , et al. Activation of the Torpedo nicotinic acetylcholine receptor. The contribution of residues αArg55 and γGlu93[J]. FEBS J, 2006, 273 (5): 960- 970. doi: 10.1111/j.1742-4658.2006.05121.x
17	HOFFMANN K , MÜLLER J S , STRICKER S , et al. Escobar syndrome is a prenatal myasthenia caused by disruption of the acetylcholine receptor fetal γ subunit[J]. Am J Hum Genet, 2006, 79 (2): 303- 312. doi: 10.1086/506257
18	BUCCAFUSCO J J . The role of central cholinergic neurons in the regulation of blood pressure and in experimental hypertension[J]. Pharmacol Rev, 1996, 48 (2): 179- 211.
19	MORGAN N V , BRUETON L A , COX P , et al. Mutations in the embryonal subunit of the acetylcholine receptor (CHRNG) cause lethal and Escobar variants of multiple pterygium syndrome[J]. Am J Hum Genet, 2006, 79 (2): 390- 395. doi: 10.1086/506256
20	SEO J , CHOI I H , LEE J S , et al. Rare cases of congenital arthrogryposis multiplex caused by novel recurrent CHRNG mutations[J]. J Hum Genet, 2015, 60 (4): 213- 215. doi: 10.1038/jhg.2015.2
21	TORRÃO A S , BRITTO L R G . Neurotransmitter regulation of neural development: acetylcholine and nicotinic receptors[J]. An Acad Bras Ciênc, 2002, 74 (3): 453- 461. doi: 10.1590/S0001-37652002000300008
22	WANG Y N , YANG W C , LI P W , et al. Myocyte enhancer factor 2A promotes proliferation and its inhibition attenuates myogenic differentiation via myozenin 2 in bovine skeletal muscle myoblast[J]. PLoS One, 2018, 13 (4): e0196255. doi: 10.1371/journal.pone.0196255
23	JUNJVLIEKE Z , KHAN R , MEI C G , et al. Effect of ELOVL6 on the lipid metabolism of bovine adipocytes[J]. Genomics, 2020, 112 (3): 2282- 2290. doi: 10.1016/j.ygeno.2019.12.024
24	李雯茜, 闫百仪, 丁恺志, 等. 离子通道调控成肌细胞分化的研究进展[J]. 生物化工, 2022, 8 (6): 167- 169.
	LI W X , YAN B Y , DING K Z , et al. Advances in myoblast differentiation and ion channel regulation[J]. Biological Chemical Engineering, 2022, 8 (6): 167- 169.
25	SI Y F , WEN H S , DU S J . Genetic mutations in jamb, jamc, and myomaker revealed different roles on myoblast fusion and muscle growth[J]. Mar Biotechnol, 2019, 21 (1): 111- 123. doi: 10.1007/s10126-018-9865-x
26	朱燕, 罗欣, 周光宏. 钙离子处理对成肌细胞μ-calpain mRNA和蛋白表达的影响[J]. 山东农业大学学报(自然科学版), 2006, 37 (4): 561-567, 572.
	ZHU Y , LUO X , ZHOU G H . Effect of cacium IoM on the MRNA and protein level expresslon of μ-calpain in rat L6 myoblast[J]. Journal of Shandong Agricultural University (Natural Science), 2006, 37 (4): 561-567, 572.
27	ROBINSON K G , VIERECK M J , MARGIOTTA M V , et al. Neuromotor synapses in Escobar syndrome[J]. Am J Med Genet A, 2013, 161 (12): 3042- 3048. doi: 10.1002/ajmg.a.36154
28	MUNIZ M M M , FONSECA L F S , MAGALHÃES A F B , et al. Use of gene expression profile to identify potentially relevant transcripts to myofibrillar fragmentation index trait[J]. Funct Integr Genomics, 2020, 20 (4): 609- 619. doi: 10.1007/s10142-020-00738-9
29	CARDANO M , TRIBIOLI C , PROSPERI E . Targeting proliferating cell nuclear antigen (PCNA) as an effective strategy to inhibit tumor cell proliferation[J]. Curr Cancer Drug Targets, 2020, 20 (4): 240- 252. doi: 10.2174/1568009620666200115162814
30	BAO B , YU X J , ZHENG W J . MiR-139-5p targeting CCNB1 modulates proliferation, migration, invasion and cell cycle in lung adenocarcinoma[J]. Mol Biotechnol, 2022, 64 (8): 852- 860. doi: 10.1007/s12033-022-00465-5
31	RAN T F , KE S , SONG X , et al. WIPI1 promotes osteosarcoma cell proliferation by inhibiting CDKN1A[J]. Gene, 2021, 782, 145537. doi: 10.1016/j.gene.2021.145537
32	宋贵兵, 贾鸿儒, 蒋蕾, 等. 秦川牛LRRN1基因表达分析及其对成肌细胞增殖分化的影响[J]. 农业生物技术学报, 2023, 31 (7): 1419- 1429.
	SONG G B , JIA H R , JIANG L , et al. Expression analysis of qinchuan cattle (Bos taurus) LRRN1 gene and its effect on proliferation and differentiation of myoblasts[J]. Journal of Agricultural Biotechnology, 2023, 31 (7): 1419- 1429.
33	LI A Q , SU X T , TIAN Y , et al. Effect of actin alpha cardiac muscle 1 on the proliferation and differentiation of bovine myoblasts and preadipocytes[J]. Animals (Basel), 2021, 11 (12): 3468.
34	ZHENG J H , VIACAVA FOLLIS A , KRIWACKI R W , et al. Discoveries and controversies in BCL-2 protein-mediated apoptosis[J]. FEBS J, 2016, 283 (14): 2690- 2700. doi: 10.1111/febs.13527
35	PEÑA-BLANCO A , GARCÍA-SÁEZ A J . Bax, bak and beyond—mitochondrial performance in apoptosis[J]. FEBS J, 2018, 285 (3): 416- 431. doi: 10.1111/febs.14186
36	TODRYK S , MELCHER A A , HARDWICK N , et al. Heat shock protein 70 induced during tumor cell killing induces Th1 cytokines and targets immature dendritic cell precursors to enhance antigen uptake[J]. J Immunol, 1999, 163 (3): 1398- 1408. doi: 10.4049/jimmunol.163.3.1398
37	LANGEN R C J , SCHOLS A M W J , KELDERS M C J M , et al. Inflammatory cytokines inhibit myogenic differentiation through activation of nuclear factor-κB[J]. FASEB J, 2001, 15 (7): 1169- 1180. doi: 10.1096/fj.00-0463
38	LONDHE P , DAVIE J K . Gamma interferon modulates myogenesis through the major histocompatibility complex class Ⅱ transactivator, CⅡTA[J]. Mol Cell Biol, 2011, 31 (14): 2854- 2866. doi: 10.1128/MCB.05397-11
39	NONG W D , HUANG F , MAO F P , et al. DCAF12 and HSPA1A may serve as potential diagnostic biomarkers for myasthenia gravis[J]. Biomed Res Int, 2022, 8587273.
40	BOLUS D J , SHANMUGAM G , NARASIMHAN M , et al. Recurrent heat shock impairs the proliferation and differentiation of C2C12 myoblasts[J]. Cell Stress Chaperones, 2018, 23 (3): 399- 410. doi: 10.1007/s12192-017-0851-4
41	ZHANG W , XUE D T , YIN H F , et al. Overexpression of HSPA1A enhances the osteogenic differentiation of bone marrow mesenchymal stem cells via activation of the Wnt/β-catenin signaling pathway[J]. Sci Rep, 2016, 6, 27622. doi: 10.1038/srep27622
42	ZHANG G H , LIU Z L , DING H , et al. Tumor induces muscle wasting in mice through releasing extracellular Hsp70 and Hsp90[J]. Nat Commun, 2017, 8 (1): 589. doi: 10.1038/s41467-017-00726-x
43	姜勇, 罗深秋. 细胞信号转导的分子基础与功能调控[M]. 北京: 科学出版社, 2005.
	JIANG Y , LUO S Q . The molecular basis and functional regulation of cellular signal transduction[M]. Beijing: Science Publishing, 2005.
44	ACOSTA-MARTINEZ M , CABAIL M Z . The PI3K/Akt pathway in meta-inflammation[J]. Int J Mol Sci, 2022, 23 (23): 15330. doi: 10.3390/ijms232315330
45	王亚宁. MEF2A对秦川牛骨骼肌成肌细胞增殖和分化的调控作用及机理研究[D]. 杨凌: 西北农林科技大学, 2019.
	WANG Y N. The roles of MEF2A in the regulation of skeletal muscle myoblasts proliferation and differentiation in Qinchuan beef cattle[D]. Yangling: Northwest A & F University, 2019. (in Chinese)

基因 Gene	转录本号 Transcript ID	引物名称 Primer name	引物序列(5'→3') Primer sequence	产物长度/bp Product length
CHRNG	NM_174273.2	CHRNG-F	CAGTCCCAGACCTACAGCAC	115
		CHRNG-R	CCCACTCCCCATTTTCTGTGA
GAPDH	NM_001034034.2	GAPDH-F	AGTTCAACGGCACAGTCAAGG	124
		GAPDH-R	ACCACATACTCAGCACCAGCA
PCNA	NM_001034494.1	PCNA-F	CCTTGGTGCAGCTAACCCTT	94
		PCNA-R	TTGGACATGCTGGTGAGGTT
CCNB1	NM_001045872.1	CCNB1-F	TACCCATTCACCATTATCAA	103
		CCNB1-R	ACTAACTATGCTGGACTACGA
CCND2	NM_001076372.1	CCND2-F	CGCCAGGTTCCATTTCA	77
		CCND2-R	CCGACAACTCCATCAAGC
CDKN1A	NM_001098958.2	CDKN1A-F	GACCAGCATGACAGATTTCTACCA	144
		CDKN1A-R	TGAAGGCCCAAGGCAAAAG
MYOD1	NM_001040478.2	MYOD1-F	AACCCCAACCCGATTTACC	196
		MYOD1-R	CACAACAGTTCCTTCGCCTCT
MYOG	NM_001111325.1	MYOG-F	GGCGTGTAAGGTGTGTAAG	85
		MYOG-R	CTTCTTGAGTCTGCGCTTCT
MYH3	NM_001101835.1	MYH3-F	TGAACGCCCTCTCCAAATCC	101
		MYH3-R	AATGAAGTGCTGTCTCGGCA
ANGPTL2	NM_001109814.1	ANGPTL2-F	GGGCACGATGAAGGTGTAGGT	114
		ANGPTL2-R	AAGGATGGCTTGGAGGGC
HSPA1A	NM_203322.3	HSPA1A-F	CTTCCGCAGACCCGCTAT	120
		HSPA1A-R	CGGTGCCCTGCCTTTT
APOD	NM_001076301.3	APOD-F	ACTCGCAAGGTAACAGAA	127
		APOD-R	CAAACCACCAGAGCAAC
CYGB	NM_001206720.2	CYGB-F	TTGAGCAGAAGGCCGAGTT	81
		CYGB-R	CCAGGAAGATGAGTCAGGGAT
PI16	NM_001024487.1	PI16-F	CGAGCGTGAGCACTACAAC	110
		PI16-R	GGGAGCCACAGCCAAT
TTR	NM_173967.3	TTR-F	GCATCCAGGACCTTGACC	115
		TTR-R	TGGCTTCCTTCCGTCTG

基因 Gene	基因ID Gene ID	差异倍数(OE) Fold change(OE)	差异倍数(sh) Fold change(sh)
TFAP2A	505849	3.44	3.44
IL36A	523429	3.57	7.29
ESM1	539571	2.30	2.62
EREG	100295476	2.20	2.61
COL13A1	613849	2.97	3.90
MYBL1	338034	2.54	2.18
NDP	511596	2.09	2.47
GPR65	788507	2.87	2.65
HSPA4L	506096	2.73	2.04
DEPDC1B	540874	2.08	2.46
RND1	508869	2.11	3.13
CXCL8	280828	4.91	3.72
PREX2	520704	3.22	2.90
HSPA1A	282254	10.78	2.73
CXCL2	281214	4.72	2.14
GRO1	281212	2.95	2.52
HSPA6	539835	695.28	3.42
H2AC6	506900	2.52	3.95

基因 Gene	基因ID Gene ID	差异倍数(OE) Fold change(OE)	差异倍数(sh) Fold change(sh)
PI16	507058	0.381	0.234
GFRA3	540009	0.290	0.497
CYGB	510299	0.402	0.474
ABCA6	537351	0.488	0.463
ABCA10	504909	0.331	0.286
COLEC12	504741	0.333	0.494
VCAM1	282118	0.181	0.377
GALNT15	618078	0.296	0.471
HVCN1	616570	0.440	0.309
CD34	281051	0.469	0.189
DDIT4L	510906	0.330	0.430
TTR	280948	0.271	0.496
ANGPTL2	512019	0.461	0.491
SLC1A7	523673	0.433	0.238
PAMR1	513841	0.308	0.266
WDR17	783416	0.242	0.238
C6	507749	0.459	0.476
SH2D3C	515820	0.350	0.433
C3	280677	0.352	0.402
ANGPT4	617915	0.272	0.220
APOD	613972	0.344	0.437
ENTPD2	100126045	0.400	0.299
ENSBTAG-00000025258	515676	0.488	0.382
ENSBTAG-00000049395	100848536	0.220	0.349
ENSBTAG-00000052577	781663	0.438	0.383

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