猪lncRNA TCONS_00791383对骨骼肌卫星细胞增殖分化的影响

doi:10.11843/j.issn.0366-6964.2020.002

摘要/Abstract

摘要： 为了探究lncRNA TCONS_00791383对猪骨骼肌卫星细胞增殖和分化的影响。本研究利用qRT-PCR技术检测出生7 d内大白仔猪6种组织（心、脾、肺、肾、背肌和腿肌）以及猪骨骼肌卫星细胞增殖分化前后TCONS_00791383的表达水平；通过设计反义核苷酸（antisense oligonucleotides，ASO）片段在猪骨骼肌卫星细胞中对TCONS_00791383进行敲低，检测敲低TCONS_00791383之后增殖分化标志基因的表达量变化；通过trans （co-expression）对TCONS_00791383进行靶基因预测，使用DAVID对其进行GO富集和KEGG通路分析。结果显示，TCONS_00791383在猪心脏中表达量最高，在脾和肾组织中不表达。在骨骼肌卫星细胞从增殖到分化的过程中，TCONS_00791383的表达量逐渐上升，且在分化后30 h表达量达到最高。在使用ASO片段敲低TCONS_00791383之后，与对照组相比，在分化24 h，增殖标志基因Pax3、Pax7表达量显著或极显著降低（P<0.05，P<0.01），分化标志基因MyoG表达量极显著降低（P<0.01），在分化48 h，增殖标志基因Pax3表达量极显著降低（P<0.01），Pax7表达量显著降低（P<0.05），分化标志基因MyHC表达量显著降低（P<0.05）。预测得到的相关靶基因富集到AMPK、ATP等多个与骨骼肌卫星细胞增殖和分化过程相关的重要信号通路。本研究表明，lncRNA TCONS_00791383可能促进猪骨骼肌卫星细胞的增殖和分化。

关键词: lncRNA TCONS_00791383, 猪, 骨骼肌卫星细胞, 增殖, 分化

Abstract: The aim of this study was to investigate the effect of lncRNA TCONS_00791383 on the proliferation and differentiation of porcine skeletal muscle satellite cells. qRT-PCR was used to detect the expression levels of TCONS_00791383 in 6 tissues (heart, spleen, lung, kidney, dorsal muscle and leg muscles) of Yorkshire piglets within 7 days of birth; At the same time, the expression levels of TCONS_00791383 before and after proliferation and differentiation of porcine skeletal muscle satellite cells were examined. The expressions of the proliferation and differentiation marker genes were examined after knocking down TCONS_00791383 in porcine skeletal muscle satellite cells using antisense oligonucleotides(ASO) fragment. Target genes of TCONS_00791383 were predicted by using trans (co-expression) method, GO enrichment and KEGG pathway analysis of these target genes were performed by DAVID. The results showed that TCONS_00791383 had the highest expression level in heart, but no expression in spleen and kidney tissues. During the process of proliferation and differentiation of skeletal muscle satellite cells, the expression level of TCONS_00791383 gradually increased and reached the peak at 30 h of differentiation. Compared with the control group, after knocking down TCONS_00791383 using the ASO fragment, the expression levels of the proliferation marker genes Pax3 and Pax7 significantly or extremely significantly decreased (P<0.05,P<0.01), and the expression level of the differentiation marker gene MyoG extremely significantly decreased (P<0.01) at 24 h of differentiation, the expression level of the proliferation marker gene Pax3 extremely significantly decreased (P<0.01) and Pax7 gene significantly decreased (P<0.05), the expression level of the differentiation marker gene MyHC significantly decreased (P<0.05) at 48 h of differentiation. The predicted target genes were enriched in AMPK, ATP and other important signaling pathways involved in the proliferation and differentiation of skeletal muscle satellite cells. This study shows that lncRNA TCONS_00791383 probably promotes the proliferation and differentiation of porcine skeletal muscle satellite cells.

Key words: lncRNA TCONS_00791383, pig, skeletal muscle satellite cells, proliferation, differentiation

中图分类号:

S828.2

李倩倩, 李龙, 黄子莹, 李长春. 猪lncRNA TCONS_00791383对骨骼肌卫星细胞增殖分化的影响[J]. 畜牧兽医学报, 2020, 51(6): 1177-1186.

LI Qianqian, LI Long, HUANG Ziying, LI Changchun. Effect of Pig lncRNA TCONS_00791383 on the Proliferation and Differentiation of Skeletal Muscle Satellite Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1177-1186.

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