基于转录组测序分析METTL14对绵羊骨骼肌卫星细胞成肌分化的影响

doi:10.11843/j.issn.0366-6964.2025.10.014

Abstract

Abstract:

The aim of this study was to explore the molecular mechanism underlying the effect of methyltransferase like 14 (METTL14) on the myogenic differentiation of sheep skeletal muscle satellite cells(SMSCs) and to provide a theoretical basis for analyzing the molecular mechanism of METTL14 regulating the differentiation of SMSCs. SMSCs were isolated and identified. The overexpression plasmid of METTL14 and the control plasmid were transfected into SMSCs, and the overexpression efficiency and the function of METTL14 affecting SMSCs differentiation were detected by RT-qPCR and Western blot. RNA-seq was performed for METTL14-overexpressed SMSCs(OE-14) and control SMSCs(OE-NC). Differentially expressed genes (DEGs) were identified under the conditions of P < 0.05 and log₂|Fold Change| > 1, GO and KEGG enrichment analysis and protein-protein interaction (PPI) network interaction analysis for DEGs was conducted. Overexpression of METTL14 markedly elevated the expression of METTL14, as well as the mRNA and protein levels of myogenic differentiation-related genes (MyHC, MyoG) in ovine SMSCs, thereby demonstrating successful METTL14 overexpression and the enhancement of SMSCs differentiation mediated by METTL14. A total of 259 differentially expressed genes were identified between the two groups. Compared with the OE-NC group, the OE-14 group had 45 up-regulated genes and 214 down-regulated genes. Differential genes were mainly enriched in nuclear pathways, Rap1 signaling pathways, etc. PPI protein network interaction analysis identified 8 hub genes (Degree≥15), including MX1, RSAD2, IFIH1, DDX58, HERC5, ISG15, MX2, and IRF7. This study indicated that METTL14 can promote the differentiation of ovine SMSCs through influencing genes and signaling pathways related to cellular processes and muscle development.

Key words: METTL14, transcriptome sequencing, skeletal muscle satellite cells, sheep, cell differentiation

CLC Number:

S826.2

HE Siqi, CHEN Qian, JIANG Lin, MA Yuehui, ZHOU Shenghua, ZHAO Qianjun. The Effect of METTL14 on Myogenic Differentiation of Ovine Skeletal Muscle Satellite Cells Based on Transcriptome Sequencing Analysis[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(10): 4925-4937.

Figures/Tables 9

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基因 Gene	引物序列(5′→3′) Primer sequence	登录号 Accession number	退火温度/℃ Annealing temperature	产物大小/bp Product size
METTL14	F：AGATTGCAGCACCTCGATCA	XM_004009592.6	60	87
	R：CCCACTTGCGTAAACACACT
MyHC	F：CTGTCCAAGTTCCGCAAGGT	XM_004010325.4	60	182
	R：GAGCTTCGTTGCACCCTCAA
MyoG	F：GAGAACTACCTGCCTGTCCAC	NM_001174109.1	60	251
	R：GCCTCGAAGGCTTCATTCAC
MX2	F：ACTGGGGCAGACAATGAGTC	NM_001078652.1	60	149
	R：GGTTGTTTTCGGACCCCTTT
RSAD2	F：TGGTTCCAGAAGTACGGTGA	XM_004005669.5	60	79
	R：TAAGGACGTTGACCTGCTCG
IFI35	F：GCGGGGACTTCAACGAAAGG	XM_004012955.6	60	182
	R：GCCTGGAGAGCCGACACAG
PGAM2	F：GTCCATCAGCAAGGAGCGT	XM_004018189.5	60	186
	R：TGCTTGACGATTCCCCGTAG
RERG	F：CAACCTACCGACACCAAGCA	XM_012175305.4	60	86
	R：CTGAATGGTGTCTTCCTGCCC
GAPDH	F：GGGTCATCATCTCTGCACCT	NM_001190390.1	60	176
	R：GGTCATAAGTCCCTCCACGA

样本 Sample	原始碱基数/bp Raw datas	过滤后碱基数/bp Clean datas	过滤后读数 Clean reads	比对率/% Ratio	Q20/%	Q30/%	GC含量/% GC content
OE-NC-1	11 617 995 000	11 478 734 663	77 185 284	95.59	98.02	94.12	48.98
OE-NC-2	11 786 511 900	11 660 724 465	78 254 396	95.51	97.97	94.01	48.81
OE-NC-3	11 790 966 300	11 668 905 187	78 291 154	95.73	97.97	94.02	49.21
OE-14-1	12 070 909 800	11 943 452 847	80 182 260	95.55	98.03	94.14	49.85
OE-14-2	11 967 072 600	11 842 913 088	79 510 606	95.52	98.03	94.12	49.12
OE-14-3	12 023 415 300	11 914 845 248	79 901 354	95.59	97.99	93.94	49.61

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The Effect of METTL14 on Myogenic Differentiation of Ovine Skeletal Muscle Satellite Cells Based on Transcriptome Sequencing Analysis

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