干扰lnc4351对牛骨骼肌卫星细胞增殖与成肌分化的影响

doi:10.16431/j.cnki.1671-7236.2019.03.013

Abstract

Abstract:

Based on the previous established isolation and culture of bovine skeletal muscle satellite cells and in vitro myogenic differentiation model,a high-throughput sequencing predicted lncRNA (lnc4351) with a great differentially expression during myogenic differentiation process was used as a target to explore the effect of lncRNA on the proliferation and differentiation of bovine skeletal muscle satellite cells,and the bioinformatics analysis and subcellular localization were performed in this study.The siRNA of lnc4351 was synthesized and transfected into bovine skeletal muscle satellite cells,the EdU staining was performed to detect the effect of interference with lnc4351 on cell proliferation.Simultaneously,an in vitro induced differentiation experiment was performed to observe the formation of myotubes after transfecting siRNA.To explore effects of interference with lnc4351 on cell differentiation,the mRNA and protein levels of the differentiation markers MyoG and MHC genes were detected using Real-time PCR and Western blotting,respectively.The results showed that lnc4351,located on chromosome 14 in cattle,was an unreported lncRNA with no protein coding potential.lnc4351 was distributed in the cytoplasm and nucleus of bovine muscle satellite cells,mainly in the nucleus.When the expression of lnc4351 was interfered,the ratio of EdU-positive cells was significantly decreased (P<0.05),suggesting that down-regulation of lnc4351 significantly inhibited the proliferation of bovine skeletal muscle satellite cells.In addition,the number of myotubes had an increasing trend and the protein levels of differentiation markers MyoG and MHC were significantly or extremely significantly up-regulated after interfering lnc4351 (P<0.05;P<0.01),indicating that down-regulation of lnc4351 could promote the myogenic differentiation process of bovine satellite cells.Down-regulating lnc4351 could inhibit the proliferation of bovine skeletal muscle satellite cells and promote its myogenic differentiation process.This study would provide a reference for further study of lncRNA on the regulation of bovine skeletal muscle development and related studies of muscle development.

Key words: lncRNA; bovine; skeletal muscle satellite cells; cell proliferation; myogenic differentiation

CLC Number:

S814.8

ZHANG Junxing, ZHU Feifei, LI Yan, CHEN Mingming, LIU Xinfeng, GUO Hong, DING Xiangbin. Effect of lnc4351 Interference on the Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells[J]. , 2019, 46(3): 747-755.

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Effect of lnc4351 Interference on the Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells

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