Skip to main content Accessibility help

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

  • Xubin Lu (a1), Hailei Xia (a1), Jingyi Jiang (a1), Xin Xu (a1), Mingxun Li (a1), Zhi Chen (a1), Yujia Sun (a2), Huimin Zhang (a1) and Zhangping Yang (a1)...


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.


Corresponding author

Author for correspondence: Zhangping Yang, Email:


Hide All
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, 16021612.
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, 1988119891.
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, 256270.
Bartel, DP (2009) MicroRNAs: target recognition and regulatory functions. Cell 136, 215233.
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, 1559215602.
Deiuliis, JA (2016) MicroRNAs as regulators of metabolic disease: pathophysiologic significance and emerging role as biomarkers and therapeutics. International Journal of Obesity 40, 88101.
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, 98
Farmer, SR (2006) Transcriptional control of adipocyte formation. Cell Metabolism 4, 263273.
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, 13151319.
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, 148154.
Friedman, RC, Farh, KK-H, Burge, CB and Bartel, DP (2009) Most mammalian mRNAs are conserved targets of microRNAs. Genome Research 19, 92105.
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, 205217.
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, 28492858.
Jagtap, S and Shivaprasad, PV (2014) Diversity, expression and mRNA targeting abilities of Argonaute-targeting miRNAs among selected vascular plants. BMC Genomics 15, 1049.
Jing, E, Gesta, S and Kahn, CR (2007) SIRT2 Regulates adipocyte differentiation through FoxO1 acetylation/deacetylation. Cell Metabolism 6, 105114.
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, 893893.
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, 123131.
Lantier, L, Williams, AS and Hughey, CC (2018) SIRT2 knockout exacerbates insulin resistance in high fat-fed mice. PLoS ONE 13, 120.
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, 29412952
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, 819831.
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, 695702.
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, 1571915727.
Lomb, DJ, Laurent, G and Haigis, MC (2010) Sirtuins regulate key aspects of lipid metabolism. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics 1804, 16521657.
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.
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, 307319.
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, 771776.
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, 119123.
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.
Rosen, ED and MacDougald, OA (2006) Adipocyte differentiation from the inside out. Nature Reviews Molecular Cell Biology 7, 885896.
Siersbaek, R, Mielsen, R and Mandrup, S (2012) Transcriptional networks and chromatin remodeling controlling adipogenesis. Trends in Endocrinology & Metabolism 23, 5664.
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, 12901297.
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, 47424753.
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, 801808.
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, 286294.
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, 429441.
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.
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, 28982906.
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, 988999.


Type Description Title
Supplementary materials

Lu et al. supplementary material
Lu et al. supplementary material

 PDF (155 KB)
155 KB

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

  • Xubin Lu (a1), Hailei Xia (a1), Jingyi Jiang (a1), Xin Xu (a1), Mingxun Li (a1), Zhi Chen (a1), Yujia Sun (a2), Huimin Zhang (a1) and Zhangping Yang (a1)...


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.