Hostname: page-component-586b7cd67f-t7fkt Total loading time: 0 Render date: 2024-11-21T11:48:13.943Z Has data issue: false hasContentIssue false
  • English
  • Français

MicroRNA-212 targets SIRT2 to influence lipogenesis in bovine mammary epithelial cell line

Published online by Cambridge University Press:  16 April 2020

Xubin Lu ,
Hailei Xia ,
Jingyi Jiang ,
Xin Xu ,
Mingxun Li ,
Zhi Chen ,
Yujia Sun ,
Huimin Zhang  and
Zhangping Yang
Xubin Lu
Affiliation:
College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu225002, China
Hailei Xia
Affiliation:
College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu225002, China
Jingyi Jiang
Affiliation:
College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu225002, China
Xin Xu
Affiliation:
College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu225002, China
Mingxun Li
Affiliation:
College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu225002, China
Zhi Chen
Affiliation:
College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu225002, China
Yujia Sun
Affiliation:
Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou225009, Jiangsu Province, China
Huimin Zhang
Affiliation:
College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu225002, China
Zhangping Yang*
Affiliation:
College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu225002, China
*
Author for correspondence: Zhangping Yang, Email: yzp@yzu.edu.cn
Get access Rights & Permissions [Opens in a new window]

Abstract

In this research paper we filter and verify miRNAs which may target silent information regulator homolog 2 (SIRT2) gene and then describe the mechanism whereby miRNA-212 might regulate lipogenic genes in mammary epithelial cell lines via targeting SIRT2. Bioinformatics analysis revealed that the bovine SIRT2 gene is regulated by three miRNAs: miR-212, miR-375 and miR-655. The three miRNAs were verified and screened by qRT-PCR, western blot, and luciferase multiplex verification techniques and only miR-212 was shown to have a targeting relationship with SIRT2. The results of co-transfecting miR-212 and silencing RNA (siRNA) showed that by targeting SIRT2, miR-212 can regulate the expression of fatty acid synthetase (FASN) and sterol regulatory element binding factor 1 (SREBP1) but not peroxisome proliferator-activated receptor gamma (PPARγ). Measurement of triglyceride (TAG) content showed that miR-212 increased the fat content of mammary epithelial cell lines. The study indicates that miR-212 could target and inhibit the expression of the SIRT2 gene to promote lipogenesis in mammary epithelial cell lines.

Keywords

LipogenesismiR-212miR-375miR-655SIRT2
Type
Research Article
Information
Journal of Dairy Research , Volume 87 , Issue 2 , May 2020 , pp. 232 - 238
Copyright
Copyright © Hannah Dairy Research Foundation 2020

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Ahn, J, Lee, H, Jung, CH, Jeon, TI and Ha, TY (2013) MicroRNA-146b promotes adipogenesis by suppressing the SIRT1-FOXO1 cascade. EMBO Molecular Medicine 5, 1602–1612.CrossRefGoogle ScholarPubMed
Armoni, M, Harel, C, Karni, S, Chen, H, Bar-Yoseph, F, Ver, MR, Quon, MJ and Karnieli, E (2006) FOXO1 Represses peroxisome proliferator-activated receptor-γ1 and -γ2 gene promoters in primary adipocytes. Journal of Biological Chemistry 281, 19881–19891.CrossRefGoogle ScholarPubMed
Aten, S, Page, CE, Kalidindi, A, Wheaton, K, Niraula, A, Godbout, JP, Hoyt, KR and Obrietan, K (2019) miR-132/212 is induced by stress and its dysregulation triggers anxiety-related behavior. Neuropharmacology 144, 256–270.CrossRefGoogle ScholarPubMed
Bartel, DP (2009) MicroRNAs: target recognition and regulatory functions. Cell 136, 215–233.CrossRefGoogle ScholarPubMed
Campos-Melo, D, Droppelmann, CA, Volkening, K and Strong, MJ (2014) Comprehensive luciferase-based reporter gene assay reveals previously masked up-regulatory effects of miRNAs. International Journal of Molecular Sciences 15, 15592–15602.CrossRefGoogle ScholarPubMed
Deiuliis, JA (2016) MicroRNAs as regulators of metabolic disease: pathophysiologic significance and emerging role as biomarkers and therapeutics. International Journal of Obesity 40, 88–101.CrossRefGoogle ScholarPubMed
Fanunza, E, Frau, A, Sgarbanti, M and Orsatti, R and Corona, A & Tramontano, E (2018) Development and validation of a novel dual luciferase reporter gene assay to quantify ebola virus VP24 inhibition of IFN signaling. Viruses-Basel 10, 98CrossRefGoogle ScholarPubMed
Farmer, SR (2006) Transcriptional control of adipocyte formation. Cell Metabolism 4, 263–273.CrossRefGoogle ScholarPubMed
Feng, D, Liu, T, Sun, Z, Bugge, A, Mullican, SE, Alenghat, T, Liu, XS and Lazar, MA (2011) A circadian rhythm orchestrated by histone deacetylase 3 controls hepatic lipid metabolism. Science (New York, N.Y.) 331, 1315–1319.CrossRefGoogle ScholarPubMed
Fernandes, T, Barretti, DL, Phillips, MI and Menezes Oliveira, E (2018) Exercise training prevents obesity-associated disorders: role of miRNA-208a and MED13. Molecular and cellular endocrinology 476, 148–154.CrossRefGoogle ScholarPubMed
Friedman, RC, Farh, KK-H, Burge, CB and Bartel, DP (2009) Most mammalian mRNAs are conserved targets of microRNAs. Genome Research 19, 92–105.CrossRefGoogle ScholarPubMed
Guo, YJ, Yu, JJ, Wang, CX, Li, K, Liu, B, Du, Y, Xiao, F, Chen, SH and Guo, FF (2017) miR-212-5p suppresses lipid accumulation by targeting FAS and SCD1. Journal of Molecular Endocrinology 59, 205–217.CrossRefGoogle ScholarPubMed
Hu, Y, Xia, W and Hou, M (2018) Macrophage migration inhibitory factor serves a pivotal role in the regulation of radiation-induced cardiac senescence through rebalancing the microRNA-34a/sirtuin 1 signaling pathway. International Journal of Molecular Medicine 42, 2849–2858.Google ScholarPubMed
Jagtap, S and Shivaprasad, PV (2014) Diversity, expression and mRNA targeting abilities of Argonaute-targeting miRNAs among selected vascular plants. BMC Genomics 15, 1049.CrossRefGoogle ScholarPubMed
Jing, E, Gesta, S and Kahn, CR (2007) SIRT2 Regulates adipocyte differentiation through FoxO1 acetylation/deacetylation. Cell Metabolism 6, 105–114.CrossRefGoogle ScholarPubMed
Kim, A-Y, Lee, E-M, Lee, E-J, Kim, J-H, Suk, K, Lee, E, Hur, K, Hong, YJ, Do, JT, Park, S and Jeong, K-S (2018) SIRT2 is required for efficient reprogramming of mouse embryonic fibroblasts toward pluripotency. Cell Death & Disease 9, 893–893.CrossRefGoogle ScholarPubMed
Kohan, K, Carvajal, R, Gabler, F, Vantman, D, Romero, C and Vega, M (2010) Role of the transcriptional factors FOXO1 and PPARG on gene expression of SLC2A4 in endometrial tissue from women with polycystic ovary syndrome. Reproduction (Cambridge, England) 140, 123–131.CrossRefGoogle ScholarPubMed
Lantier, L, Williams, AS and Hughey, CC (2018) SIRT2 knockout exacerbates insulin resistance in high fat-fed mice. PLoS ONE 13, 1–20.CrossRefGoogle ScholarPubMed
Lefterova, MI, Zhang, Y, Steger, DJ, Schupp, M, Schug, J, Cristancho, A, Feng, D, Zhuo, D, Stoeckert, CJ Jr, Liu, XS and Lazar, MA (2008) PPARgamma and C/EBP factors orchestrate adipocyte biology via adjacent binding on a genome wide scale. Genes & Development 22, 2941–2952CrossRefGoogle Scholar
Lei, L, Zhou, C, Yang, X and Li, LP (2018) Down-regulation of microRNA-375 regulates adipokines and inhibits inflammatory cytokines by targeting AdipoR2 in non-alcoholic fatty liver disease. Clinical And Experimental Pharmacology And Physiology 45, 819–831.CrossRefGoogle ScholarPubMed
Li, DD, Bai, L, Wang, TT, Xie, Q, Chen, ML, Fu, YT and Wen, Q (2018) Function of miR-212 as a tumor suppressor in thyroid cancer by targeting SIRT1. Oncology Reports 39, 695–702.Google ScholarPubMed
Lin, L, Wang, Z, Jin, H, Shi, H, Lu, Z and Qi, Z (2016) MiR-212/132 is epigenetically downregulated by SOX4/EZH2-H3K27me3 feedback loop in ovarian cancer cells. Biology and Medicine 37, 15719–15727.Google Scholar
Lomb, DJ, Laurent, G and Haigis, MC (2010) Sirtuins regulate key aspects of lipid metabolism. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics 1804, 1652–1657.CrossRefGoogle ScholarPubMed
Mestdagh, P, Van Vlierberghe, P, De Weer, A, Muth, D, Westermann, F, Speleman, F and Vandesompele, J (2009) A novel and universal method for microRNA RT-qPCR data normalization. Genome Biology 10, R64.CrossRefGoogle ScholarPubMed
Perrini, S, Porro, S, Nigro, P, Cignarelli, A, Caccioppoli, C, Genchi, VA, Martines, G, Fazio, MD, Capuano, P, Natalicchio, A, Laviola, L and Giorgino, F (2020) Reduced SIRT1 and SIRT2 expression promotes adipogenesis of human visceral adipose stem cells and associates with accumulation of visceral fat in human obesity. International Journal of Obesity 44, 307–319.CrossRefGoogle ScholarPubMed
Picard, F, Kurtev, M, Chung, N, Topark-Ngarm, A, Senawong, T, Machado De Oliveira, R, Leid, M, McBurney, MW and Guarente, L (2004) Sirt1 promotes fat mobilization in white adipocytes by repressing PPAR-gamma. Nature 429, 771–776.CrossRefGoogle ScholarPubMed
Ran, L and Eveline, MI-A (2017) Altered gene expression of epigenetic modifying enzymes in response to dietary supplementation with linseed oil. Journal of Dairy Research 84, 119–123.Google Scholar
Ronn, T, Volkov, P, Davegardh, C, Dayeh, T, Hall, E, Olsson, AH, Nilsson, E, Tornberg, A, Nitert, MD, Eriksson, KF, Jones, HA, Groop, L and Ling, C (2013) A six months exercise intervention influences the genome-wide DNA methylation pattern in human adipose tissue. PLoS Genetics 9, e1003572.CrossRefGoogle ScholarPubMed
Rosen, ED and MacDougald, OA (2006) Adipocyte differentiation from the inside out. Nature Reviews Molecular Cell Biology 7, 885–896.CrossRefGoogle ScholarPubMed
Siersbaek, R, Mielsen, R and Mandrup, S (2012) Transcriptional networks and chromatin remodeling controlling adipogenesis. Trends in Endocrinology & Metabolism 23, 56–64.CrossRefGoogle ScholarPubMed
Wagschal, A, Najafi-Shoushtari, SH, Wang, L, Goedeke, L, Sinha, S, deLemos, AS, Black, JC, Ramirez, CM, Li, YX, Tewhey, R, Hatoum, I, Shah, N, Lu, Y, Kristo, F, Psychogios, N, Vrbanac, V, Lu, YC, Hla, T, de Cabo, R, Tsang, JS, Schadt, E, Sabeti, PC, Kathiresan, S, Cohen, DE, Whetstine, J, Chung, RT, Fernandez-Hernando, C, Kaplan, LM, Bernards, A, Gerszten, RE and Naar, AM (2015) Genome wide. Nature Medicine 21, 1290–1297.CrossRefGoogle ScholarPubMed
Wanet, A, Tacheny, A, Arnould, T and Renard, P (2012) miR-212/132 expression and functions: within and beyond the neuronal compartment. Nucleic Acids Research 40, 4742–4753.CrossRefGoogle ScholarPubMed
Wang, F and Tong, Q (2009) SIRT2 suppresses adipocyte differentiation by deacetylating FOXO1 and enhancing FOXO1's repressive interaction with PPAR gamma. Molecular Biology of the Cell 20, 801–808.CrossRefGoogle Scholar
Xie, MX, Fu, ZY, Cao, JX, Liu, Y, Wu, J, Li, Q and Chen, Y (2018) MicroRNA-132 and microRNA-212 mediate doxorubicin resistance by down-regulating the PTEN-AKT/NF-kappa B signaling pathway in breast cancer. Biomedicine & Pharmacotherapy 102, 286–294.CrossRefGoogle ScholarPubMed
Xu, L, Wang, L and Zhou, LX (2019 a) The SIRT2/cMYC pathway inhibit peroxidation-related apoptosis in cholangiocarcinoma through metabolic reprogramming. Neoplasia (New York, N.Y.) 21, 429–441.CrossRefGoogle Scholar
Xu, WW, Chen, QM, Jia, YH, Deng, JX, Jiang, SQ, Qin, GS, Qiu, QQ, Wang, XP, Yang, XR and Jiang, HS (2019 b) Isolation, characterization, and SREBP1 functional analysis of mammary epithelial cell in buffalo. Journal of Food Biochemistry 00, e12997.Google Scholar
Yang, WC, Guo, WL, Zan, LS, Wang, YN and Tang, KQ (2017) Bta-miR-130a regulates the biosynthesis of bovine milk fat by targeting peroxisome proliferator activated receptor gamma. Journal of Animal Sciences 95, 2898–2906.Google ScholarPubMed
Zhang, W, Sun, Q, Zhong, W, Sun, X and Zhou, Z (2016) Hepatic peroxisome proliferator activated receptor gamma signaling contributes to alcohol induced hepatic steatosis and inflammation in mice. Alcoholism, Clinical and Experimental Research 40, 988–999.CrossRefGoogle ScholarPubMed
Supplementary material: PDF

Lu et al. supplementary material

Lu et al. supplementary material

Download Lu et al. supplementary material(PDF)
PDF 155.2 KB