猪C1QTNF3基因可变剪接体的鉴定及介导miR-101调控成脂分化的研究

doi:10.11843/j.issn.0366-6964.2021.011.006

Abstract

Abstract: The study aimed to explore the alternative splicing isoforms of pig C1QTNF3 gene and mechanism of miR-101 targeting C1QTNF3 expression in promoting porcine adipose-derived stromal vascular cells (ASVC) adipogenesis. The heart, liver, spleen, lung, kidney, stomach, biceps femoris, psoas magnus, abdominal fat and back fat were obtained from 3 30-day-old healthy Mashen male piglets. The full-length coding sequence (CDS) of C1QTNF3 transcripts were cloned by RT-PCR, the biological characteristics of C1QTNF3 protein were analyzed by bioinformatics methods. The expression patterns of C1QTNF3 transcripts in porcine tissues were detected by qRT-PCR. Bioinformatics prediction revealed that miR-101 was the upstream regulatory factor of C1QTNF3. The effect of miR-101 on C1QTNF3 was verified by dual luciferase reporter assay. The effect of overexpression of miR-101 on adipogenesis of porcine ASVC was detected by Oil Red O staining and qRT-PCR. Two transcripts of C1QTNF3 gene, C1QTNF3 and C1QTNF3-1, were successfully obtained in this study. C1QTNF3-1 was a newly identified transcript, which lacked 219 bp (73 amino acids) in the first exon compared with C1QTNF3. The phylogenetic tree analysis showed that the porcine C1QTNF3 and C1QTNF3-1 proteins sequence were highly related to Homo sapiens and Manis javanica. C1QTNF3 and C1QTNF3-1 were expressed in all porcine tissues, the expression level of C1QTNF3-1 was significantly higher than that of C1QTNF3 (P<0.01). Overexpression of miR-101 significantly reduced the relative luciferase activity of C1QTNF3 3'UTR(P<0.01) and mRNA expression of C1QTNF3(P<0.01); the mRNA expression of key genes for adipogenesis, PPARγ, C/EBPβ, SREBP-1c and FABP4 were significantly increased (P<0.01). In this study, two transcripts of pig C1QTNF3 gene were successfully cloned. The expression of C1QTNF3-1 was significantly higher than C1QTNF3 in different porcine tissues, suggesting it was the predominant transcript. In addition, miR-101 was identified as the upstream regulatory factor of C1QTNF3. By reducing C1QTNF3 gene expression, miR-101 promoted porcine ASVC adipogenesis, which enriched the biological role and regulatory network of C1QTNF3.

Key words: pig, C1QTNF3 gene, adipogenesis, miR-101, alternative splicing

CLC Number:

S828.2

YANG Yang, ZHANG Xuelian, ZHANG Wanfeng, LI Wenxia, LI Wenxin, CAI Chunbo, LU Chang, GAO Pengfei, GUO Xiaohong, LI Bugao, CAO Guoqing. Study of Alternative Splicing of Porcine C1QTNF3 Gene and Its Regulation on Adipogenesis by Mediating miR-101[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(11): 3042-3052.

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Study of Alternative Splicing of Porcine C1QTNF3 Gene and Its Regulation on Adipogenesis by Mediating miR-101

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