干扰MAPK6基因对秦川牛成肌细胞分化的影响研究

doi:10.11843/j.issn.0366-6964.2025.11.011

Abstract

Abstract:

The aim of this study was to investigate the effect of interfering with MAPK6 (extracellular signal regulated kinase 3, ERK3) gene on the differentiation of Qinchuan cattle myoblasts and its action mechanism. The expression profile of the MAPK6 gene in different tissues of Qinchuan cattle was detected using qRT-PCR technology. siRNA technology was used to interfere with the expression of MAPK6 gene in Qinchuan cattle myoblasts. The interference efficiency and expression levels of myotube differentiation related genes were detected by qRT-PCR and Western blot, and changes in myotube phenotype were observed. Subsequently, transcriptome sequencing was performed to screen differentially expressed genes, followed by GO functional annotation and KEGG pathway enrichment analysis. Additionally, preliminary validation of myogenic differentiation-related pathways was conducted. The results indicated that MAPK6 was highly expressed in the longissimus dorsi muscle tissue of Qinchuan cattle. Interfering with MAPK6 significantly promoted myotube differentiation and fusion in Qinchuan cattle myoblasts (P < 0.01), significantly increased the expression of myogenic differentiation-related genes MEF2C and MYH7 (P < 0.01) and the protein expression of MEF2C and MYL2 (P < 0.01), and significantly increased the protein expression of MYH7 (P < 0.05). The transcriptome results showed that, compared with the control group, there were 3 224 differentially expressed genes after interfering with MAPK6, of which 1 707 were upregulated and 1 517 were downregulated. The KEGG enrichment results showed that differential genes were mainly enriched in the PI3K-AKT and MAPK signaling pathways. Interfering with the MAPK6 gene could significantly reduces the expression of ERK1/2 and p-ERK1/2 proteins (P < 0.01) in the MAPK signaling pathway. This study preliminarily elucidates the role and mechanism of MAPK6 gene in beef muscle production, laying a foundation for improving beef quality and guiding molecular breeding strategies in beef cattle.

Key words: Qinchuan cattle, skeletal muscles, MAPK6 gene, myoblast differentiation

CLC Number:

S823.2

LI Bingzhi, GUO Juntao, WANG Jianfang, YU Shengchen, PAN Yueting, YU Hengwei, LIU Haibing, ZHANG Ke, CHENG Gong, TIAN Wanqiang, ZAN Linsen. Study on the Effect of Interfering with MAPK6 Gene on the Differentiation of Qinchuan Cattle Myoblast Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(11): 5475-5488.

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基因 Gene	GenBank号 GenBank number	引物 Primer	引物序列(5′→3′) Primer sequence	产物长度/bp Product length
GAPDH	NM_001034034.2	GAPDH-F	AGTTCAACGGCACAGTCAAGG	124
GAPDH	NM_001034034.2	GAPDH-R	ACCACATACTCAGCACCAGCA	124
MAPK6	XM_059890855.1	MAPK6-F	ACTTGGTGCTGAAGATAGGTGA	206
MAPK6	XM_059890855.1	MAPK6-R	AAAGAGGGTTTTACCAGTCAGC	206
MEF2C	XM_059888100.1	MEF2C-F	GCACCAACAAGCTGTTCCAG	117
MEF2C	XM_059888100.1	MEF2C-R	GAACAGCCTCCACGATGTCT	117
MYH7	XM_024997290.2	MYH7-F	TCCAGTCTTCTCTCCTTGCTG	187
MYH7	XM_024997290.2	MYH7-R	AGGTCGAAAGGCCTGGTCTG	187
MYL2	NM_001035025.2	MYL2-F	CTTTCCACCATGTCACCTAAGA	114
MYL2	NM_001035025.2	MYL2-R	GATGGTGAAGGCCTCCTTAAA	114
CEND1	NM_001080222.1	CEND1-F	GAGCAGCAGCGCCCATA	125
CEND1	NM_001080222.1	CEND1-R	CTTCCCTCTGGACTCCATGC	125
SORD	NM_001037320.1	SORD-F	CTGGCAGCATGGTCGAATTG	102
SORD	NM_001037320.1	SORD-R	TGCCTGACCAATGATCCCAC	102
WASF3	XM_005213794.5	WASF3-F	CCAGGTTATCACAGAGCGCA	102
WASF3	XM_005213794.5	WASF3-R	CCGGGTACGAGTAATCCGTG	102
VPS50	NM_001105413.2	VPS50-F	ATGAGGGTCGTGCCTTGATG	162
VPS50	NM_001105413.2	VPS50-R	TGTGCTCTTTGATCCAGCGT	162
ZFP90	XM_059877474.1	ZFP90-F	CGAGTGAAGACGGCACAGAA	85
ZFP90	XM_059877474.1	ZFP90-R	CCAGGGACAGGGTTGACTTC	85
CCNG1	NM_001013364.1	CCNG1-F	GCTGTGAAGCCTCAAAACATC	170
CCNG1	NM_001013364.1	CCNG1-R	TCCAGGCTTGTCATTTGCATT	170
NR1D2	XM_059882241.1	NR1D2-F	GTCTAATTACCTGGTTTCCCCC	127
NR1D2	XM_059882241.1	NR1D2-R	GACGCGGACTGGAAGCTATT	127
GCSH	NM_174844.1	GCSH-F	TATTGAGGAGTGAAACTGGAACCC	188
GCSH	NM_174844.1	GCSH-R	AGATGCAGTGTTTATCTGAAGTCAT	188
ABCB10	XM_024986813.2	ABCB10-F	CTCATGCAGACGTCAGGTCA	107
ABCB10	XM_024986813.2	ABCB10-R	TCTCCTGTGCGTGTCTTGTC	107

样品 Sample	原始数据Raw data		过滤数据Clean data		有效比对率/% Valid ratio (reads)	Q20%	Q30%	GC含量/% GC content
样品 Sample	读数 Read	碱基/G Base	读数 Read	碱基/G Base	有效比对率/% Valid ratio (reads)	Q20%	Q30%	GC含量/% GC content
MAPK6S1	40 273 634	6.04	38 754 750	5.81	96.23	99.93	98.04	50.50
MAPK6S2	40 890 852	6.13	39 414 348	5.91	96.39	99.94	97.97	50.50
MAPK6S3	40 659 854	6.10	39 237 220	5.89	96.50	99.94	98.04	49.50
SINC1	39 899 744	5.98	38 249 602	5.74	95.86	99.93	97.97	50.50
SINC2	38 885 912	5.83	37 072 804	5.56	95.34	99.92	97.89	51.50
SINC3	39 628 850	5.94	38 159 344	5.70	96.29	99.93	97.97	51.50

样品 Sample	过滤数据 Clean reads	比对读数 Mapped reads	唯一比对读数 Unique mapped reads	多比对读数 Multi mapped reads
MAPK6S1	38 754 750	36 246 759(93.53%)	35 413 870(91.38%)	832 889(2.15%)
MAPK6S2	39 414 348	36 937 794(93.72%)	36 105 382(91.60%)	832 412(2.11%)
MAPK6S3	39 237 220	36 907 942(94.06%)	36 079 312(91.95%)	828 630(2.11%)
SINC1	38 249 602	35 839 948(93.70%)	34 934 213(91.33%)	905 735(2.37%)
SINC2	37 072 804	34 373 945(92.72%)	33 513 528(90.40%)	860 417(2.32%)
SINC3	38 159 344	35 644 691(93.41%)	34 805 597(91.21%)	839 094(2.20%)

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Study on the Effect of Interfering with MAPK6 Gene on the Differentiation of Qinchuan Cattle Myoblast Cells

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