基于RNA-seq技术分析microRNA 29a对八眉猪仔猪小肠上皮细胞基因表达的影响

doi:10.3969/j.issn.1674-7968.2022.08.010

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Abstract Small intestinal epithelial cells (IECs) are important basis for the small intestine to play an important role in digestion and absorption of nutrients and intestinal barrier. microRNA 29a (miR-29a) plays an important role in cell apoptosis, differentiation and proliferation during animal growth and development. In order to clarify the effect of miR-29a on the gene expression of small intestinal epithelial cells of piglets (Sus scrofa), miR-29a mimics group, miR-29a inhibitor group, and negative control group were constructed. The IECs were collected after transfection 48 h, extracted RNA to construct libraries and performed Illumina sequencing; The differential expressed genes (DEGs) were screened and subjected to GO functional annotation and KEGG pathway analysis; Venn analysis was used to screen target differential genes and construct target gene co-expression network, 4 genes were selected randomly to verify the accuracy of transcriptome data by qPCR. A total of 9 transcriptome libraries were obtained in this study and total clean base was 55.42 Gb, and the comparison rate with the reference genome reached more than 95%. The DESeq2 software was used to analyze the expressed genes obtained by sequencing, and the results showed that 7 102 DEGs were expressed between the miR-29a inhibitor group (VPA) and the negative control group (VPC), of which 2 558 up-regulated genes and 3 544 down-regulated genes were found in VPA; there were 2 829 DEGs between the two groups, of which 1 563 genes were up-regulated genes in miR-29a mimics group (VPT), and 1 266 genes were down-regulated in the VPT; there were 503 DEGs in the VPA and VPT. There were 197 up-regulated and 306 down-regulated genes in VPA. The GO functional annotation and KEGG pathway enrichment analysis of the differential genes showed that the IECs after miR-29a treatment were in a very active stage of cell proliferation and differentiation and the obtained DEGs were mainly enriched in DNA replication and cell cycle pathways. qPCR results were consistent with the sequencing data. The above results suggested that miR-29a might be regulated cell apoptosis by affecting genes such as CDKN2C, E2F1, MCM family and POLE in IECs, thereby affecting intestinal absorption and intestinal barrier function. This study provides basic for studying the effect of miR-29a on the intestinal of Bamei piglets.

Key words： MicroRNA 29a Transcriptome Bamei Small intestine

Received: 15 November 2021

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Corresponding Authors: *jiajianlei87@163.com

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	XIE Wen
	REN Hao
	WEI Tao
	ZHANG Ying-Ying
	ZHANG Huai-Xia
	CHEN Qian
	JIA Jian-Lei

Cite this article:

XIE Wen,REN Hao,WEI Tao, et al. Analysis of the Effect of microRNA 29a on Gene Expression in Small Intestinal Epithelial Cells of Bamei Piglets (Sus scrofa) Based on RNA-seq Technology[J]. 农业生物技术学报, 2022, 30(8): 1547-1558.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2022.08.010 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2022/V30/I8/1547

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