PFN1-PTEN通过调控PI3K/AKT/mTOR抑制牛骨骼肌卫星细胞分化

doi:10.11843/j.issn.0366-6964.2025.11.012

Abstract

Abstract:

This study aimed to investigate the effect of PFN1 on the myogenic differentiation of bovine skeletal muscle satellite cells (BSMSCs) through its interaction with PTEN. Small interfering RNA (siRNA) and overexpression plasmids were transfected into primary bovine skeletal muscle satellite cells isolated from fetal bovine leg muscles, with 3 biological replicates for each treatment. The differentiation process of the cells was observed under an optical microscope. RNA and protein samples were collected at specific time points during the differentiation phase for analysis by quantitative real-time PCR (qRT-PCR) and Western blot. PFN1 knockdown promoted the formation of more robust myotubes, with significantly increased MyHC mRNA and protein levels. Conversely, PFN1 overexpression reduced myotube formation (thinner and fewer) and significantly decreased MyHC expression. Co-IP confirmed the interaction between PFN1 and PTEN, and PFN1 overexpression increased PTEN protein levels. Under optical microscope observation, PTEN knockdown resulted in fewer myotubes, with elevated MyHC and MyOG mRNA levels but reduced MyHC protein levels. Key proteins in the PI3K/AKT/mTOR pathway were downregulated, while p-mTOR levels increased. PFN1 knockdown upregulated key proteins in the PI3K/AKT/mTOR pathway, whereas PFN1 overexpression downregulated them. PFN1 interacts with PTEN to inhibit the PI3K/AKT/mTOR pathway, thereby negatively regulating BSMSCs myogenic differentiation. This study provides new insights into the molecular regulatory network of muscle development.

Key words: bovine, skeletal muscle satellite cells, PFN1, myogenic differentiation, PI3K/AKT/mTOR

CLC Number:

S823.2

ZHANG Shuai, XU Jing, YANG Peihong, GUO Yiwen, HU Debao, LI Xin, DING Xiangbin, GUO Hong, ZHANG Linlin. PFN1-PTEN Inhibits Bovine Skeletal Muscle Satellite Cell Differentiation by Regulating the PI3K/AKT/mTOR Pathway[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(11): 5489-5501.

Figures/Tables 11

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基因Gene	引物序列(5′→3′) Sequence	产物大小/bp Product size
PFN1 forward	TCGTGAACGGGCTAACACTT	171
PFN1 reverse	CTTTGCCCATCAGCAGGACT
GAPDH forward	GAAGGTCGGAGTGAACGGAT	91
GAPDH reverse	ATTGATGGCGACGATGTCCA
PTEN forward	TATTCTTCTCCCCATTCCGCTGC	174
PTEN reverse	AATGGCTCTGGACTTGGCGG
MyoG forward	GGCTGACAAATGCCAGACTATCC	140
MyoG reverse	TGGTCCCTTGCTTTATCTCCCT
MyHC forward	CTGGAATCCGGAGGCAGAA	105
MyHC reverse	TTTTCGAAGGTAGGGAGCGG

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PFN1-PTEN Inhibits Bovine Skeletal Muscle Satellite Cell Differentiation by Regulating the PI3K/AKT/mTOR Pathway

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