Proteome-wide Screening of Transcription Factor DNA Binding Activity in HepG2 Cells after Oleic Acid Treatment

doi:10.13523/j.cb.20150504

China Biotechnology

2015, Vol. 35

Issue (5): 22-31 DOI: 10.13523/j.cb.20150504

Proteome-wide Screening of Transcription Factor DNA Binding Activity in HepG2 Cells after Oleic Acid Treatment

LIANG Li-zhu^1,2,3, SUN Jia-nan^3,4, LI Kai^3,5, LIU Ming-wei³, DING Chen^2,3, QIN Jun^1,2,3

1. Graduate School of Anhui Medical University, Hefei 230032, China;
2. Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China;
3. Beijing Proteome Research, State Key Laboratory of Proteomics, Beijing 102206, China;
4. College of Pharmacy of Shenyang Pharmaceutical University, Shenyang 110016, China;
5. School of Life Sciences of Hebei United University, Tangshan 063009, China

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Abstract

Nonalcoholic fatty liver disease (NAFLD), one of the most prevalent chronic diseases worldwide, is associated with accumulation of fat in liver and is a manifestation of metabolic syndromes (including obesity, diabetes, and insulin resistance). HepG2 cells are selected to establish a model in vitro about hepatic steatosis and to identify the transcription factors (TFs) that change their binding affinity to DNA in response to hepatic steatosis. HepG2 cells are incubated with 2mmol/L oleic acid for 24 hours. Oil-Red-O staining is done to show triglycerides (TG) and assess the extent of hepatic steatosis. The nuclear extraction from each group is separated from the cytoplasmic extraction and the transcription factors in nuclear extraction are enriched by a concatenated tandem array of consensus transcription factor response element (cat TFRE). By combination of catTFRE and an in-depth analysis of proteomics expression profiling, the dynamic description and quantitative analysis of the transcription factors' DNA binding activity changes are shown. Then the functions of the altered transcription factors are analyzed by IPA(integrated pathway analysis). 170 TFs are identified in the control groups and 190 TFs are identified in the oleic acid treated groups. 208TFs are identified in all experiments. In the 208 TFs, DNA binding activity of 67 TFs are up-regulated obviously and 34 TFs are down-regulated obviously. The DNA binding activity of MLX and MLXIPL, which play important roles in glycometabolism, down-regulated obviously. The DNA binding activity of NF-κB1 is up-regulated obviously and suggested that the NF-κB-mediated inflammation is activated.The results of IPA revealed that NRF2-mediated oxidative stress response is activated in response to the oxidative stress and the progresses of gene expression, cell proliferation, cell differentiation and cell cycle are increased. The conclusion is that after treated with oleic acid, although the balance between glycometabolism and lipid metabolism is broken, inflammation is activated and oxidative damage is caused, HepG2 cells tend to take actions like increasing gene expression, cell proliferation and activating oxidative stress response at the transcription level and finaly keep cells survival.

Key words： Proteome Transcription factors HepG2 cells Oleic acid Steatosis

Received: 30 January 2015 Published: 25 May 2015

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LIANG Li-zhu, SUN Jia-nan, LI Kai, LIU Ming-wei, DING Chen, QIN Jun. Proteome-wide Screening of Transcription Factor DNA Binding Activity in HepG2 Cells after Oleic Acid Treatment. China Biotechnology, 2015, 35(5): 22-31.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150504 OR https://manu60.magtech.com.cn/biotech/Y2015/V35/I5/22

[1] Evans R M,Barish G D, Wang Y X. PPARs and the complex journey to obesity. Nature Medicine,2004,10(4):355-361.

[2] Schaffer J E. Lipotoxicity: when tissues overeat. Current Opinion in Lipidology,2003,14(3):281-287.

[3] Adams L A,Angulo PLindor K D. Nonalcoholic fatty liver disease. Canadian Medical Association Journal,2005,172(7):899-905.

[4] Adams L A,Lymp J F,St Sauver J, et al. The natural history of nonalcoholic fatty liver disease: a population-based cohort study. Gastroenterology,2005,129(1):113-121.

[5] Crabb D W,Galli A,Fischer M, et al. Molecular mechanisms of alcoholic fatty liver: role of peroxisome proliferator-activated receptor alpha. Alcohol,2004,34(1):35-38.

[6] Cohen J C,Horton J D,Hobbs H H. Human fatty liver disease: old questions and new insights. Science,2011,332(6037):1519-1523.

[7] Sunny N E,Parks E J,Browning J D, et al. Excessive hepatic mitochondrial TCA cycle and gluconeogenesis in humans with nonalcoholic fatty liver disease. Cell Metabolism,2011,14(6):804-810.

[8] Guo Y,Darshi M,Ma Y, et al. Quantitative proteomic and functional analysis of liver mitochondria from high fat diet (HFD) diabetic mice. Molecular & Cellular Proteomics,2013,12(12):3744-3758.

[9] Blake W L,Clarke S D. Suppression of rat hepatic fatty acid synthase and S14 gene transcription by dietary polyunsaturated fat. The Journal of Nutrition,1990,120(12):1727-1729.

[10] Jump D B,Clarke S D. Regulation of gene expression by dietary fat. Annu Rev Nutr,1999,19(1):63-90.

[11] Jump D B,Clarke S D,MacDougald O, et al. Polyunsaturated fatty acids inhibit S14 gene transcription in rat liver and cultured hepatocytes. Proceedings of the National Academy of Sciences of the United States of America,1993,90(18):8454-8458.

[12] Vaquerizas J M,Kummerfeld S K,Teichmann S A, et al. A census of human transcription factors: function, expression and evolution. Nature Reviews Genetics,2009,10(4):252-263.

[13] Ding C,Chan D W,Liu W, et al. Proteome-wide profiling of activated transcription factors with a concatenated tandem array of transcription factor response elements. Proceedings of the National Academy of Sciences of the United States of America,2013,110(17):6771-6776.

[14] Jump D B. Fatty acid regulation of hepatic lipid metabolism. Current Opinion in Clinical Nutrition and Metabolic Care,2011,14(2):115-120.

[15] Ren B,Thelen A P,Peters J M, et al. Polyunsaturated fatty acid suppression of hepatic fatty acid synthase and S14 gene expression does not require peroxisome proliferator-activated receptor alpha. The Journal of Biological Chemistry,1997,272(43):26827-26832.

[16] Wang Y,Botolin D,Xu J, et al. Regulation of hepatic fatty acid elongase and desaturase expression in diabetes and obesity. Journal of Lipid Research,2006,47(9):2028-2041.

[17] Postic C,Girard J. Contribution of de novo fatty acid synthesis to hepatic steatosis and insulin resistance: lessons from genetically engineered mice. The Journal of Clinical Investigation,2008,118(3):829-838.

[18] Lee J Y,Plakidas A,Lee W H, et al. Differential modulation of toll-like receptors by fatty acids preferential inhibition by n-3 polyunsaturated fatty acids. Journal of Lipid Research,2003,44(3):479-486.

[19] Ribeiro P S,Cortez-Pinto H,Sola S, et al. Hepatocyte apoptosis, expression of death receptors, and activation of NF-kappaB in the liver of nonalcoholic and alcoholic steatohepatitis patients. The American Journal of Gastroenterology,2004,99(9):1708-1717.

[20] Jump D B,Tripathy S,Depner C M. Fatty acid regulated transcription factors in the liver. Annual Review of Nutrition,2013,33(4):249-249.

[21] Gu L Y,Qiu L W,Chen X F, et al. Oleic acid induced hepatic steatosis is coupled with downregulation of aquaporin 3 and upregulation of aquaporin 9 via activation of p38 signaling. Hormone and Metabolic Research,2014,13(3):125-129.

[22] Hwang Y J,Wi H R,Kim H R, et al. Pinus densiflora Sieb. et Zucc. alleviates lipogenesis and oxidative stress during oleic acid-induced steatosis in HepG2 cells. Nutrients,2014,6(7):2956-2972.

[23] Holzer R G,Park E J,Li N, et al. Saturated fatty acids induce c-Src clustering within membrane subdomains, leading to JNK activation. Cell,2011,147(1):173-184.

[24] Listenberger L L,Han X,Lewis S E, et al. Triglyceride accumulation protects against fatty acid-induced lipotoxicity. Proceedings of the National Academy of Sciences,2003,100(6):3077-3082.

[25] Wu H T,Chen W,Cheng K C, et al. Oleic acid activates peroxisome proliferator-activated receptor δ to compensate insulin resistance in steatotic cells. The Journal of Nutritional Biochemistry,2012,23(10):1264-1270.

[26] Chen Y,Billadello J J,Schneider D J. Identification and localization of a fatty acid response region in the human plasminogen activator inhibitor-1 gene. Arteriosclerosis, Thrombosis, and Vascular Biology,2000,20(12):2696-2701.

[27] Byon C H,Hardy R W,Ren C, et al. Free fatty acids enhance breast cancer cell migration through plasminogen activator inhibitor-1 and SMAD4. Laboratory Investigation; A Journal of Technical Methods and Pathology,2009,89(11):1221-1228.

[28] Reyes-Quiroz M E,Alba G,Saenz J, et al. Oleic acid modulates mRNA expression of liver X receptor (LXR) and its target genes ABCA1 and SREBP1c in human neutrophils. European Journal of Nutrition,2014,7(3):1-11.

[29] Zor T,Selinger Z. Linearization of the bradford protein assay increases its sensitivity: theoretical and experimental studies. Analytical Biochemistry,1996,236(2):302-308.

[30] Schwanhausser B,Busse D,Li N, et al. Global quantification of mammalian gene expression control. Nature,2011,473(7347):337-342.

[31] Arkan M C,Hevener A L,Greten F R, et al. IKK-beta links inflammation to obesity-induced insulin resistance. Nature Medicine,2005,11(2):191-198.

[32] Cai D,Yuan M,Frantz D F, et al. Local and systemic insulin resistance resulting from hepatic activation of IKK-beta and NF-kappaB. Nature Medicine,2005,11(2):183-190.

[33] Dietze R,Konrad L,Shihan M, et al. Cardiac glycoside ouabain induces activation of ATF-1 and StAR expression by interacting with the alpha4 isoform of the sodium pump in Sertoli cells. Biochimica et Biophysica Acta,2013,1833 (3):511-519.

[34] Gan B,Lim C,Chu G, et al. FoxOs enforce a progression checkpoint to constrain mTORC1-activated renal tumorigenesis. Cancer cell,2010,18(5):472-484.

[35] Lu M,Lawrence D A,Marsters S, et al. Opposing unfolded-protein-response signals converge on death receptor 5 to control apoptosis. Science,2014,345(6192):98-101.

[36] Kaufman R J, Cao S. Inositol‐requiring 1/X‐box‐binding protein 1 is a regulatory hub that links endoplasmic reticulum homeostasis with innate immunity and metabolism. EMBO Molecular Medicine,2010,2(6):189-192.

[37] Verfaillie T,Salazar M,Velasco G, et al. Linking ER stress to autophagy: potential implications for cancer therapy. International Journal of Cell Biology,2010,7(4):352-356.

[38] Klune J R,Dhupar R,Cardinal J, et al. HMGB1: endogenous danger signaling. Molecular Medicine,2008,14(7):476-476.

[39] Ohmori H,Luo Y,Kuniyasu H. Non-histone nuclear factor HMGB1 as a therapeutic target in colorectal cancer. Expert Opinion on Therapeutic Targets,2011,15(2):183-193.

[40] Andres-Hernando A,Lanaspa M A,Rivard C J, et al. Nucleoporin 88 (Nup88) is regulated by hypertonic stress in kidney cells to retain the transcription factor tonicity enhancer-binding protein (TonEBP) in the nucleus. Journal of Biological Chemistry,2008,283(36):25082-25090.

[41] Hofmanova J,Vaculova A,Kozubik A. Polyunsaturated fatty acids sensitize human colon adenocarcinoma HT-29 cells to death receptor-mediated apoptosis. Cancer Letters,2005,218(1):33-41.

[42] Schley P D,Brindley D N,Field C J. (n-3) PUFA alter raft lipid composition and decrease epidermal growth factor receptor levels in lipid rafts of human breast cancer cells. The Journal of Nutrition,2007,137(3):548-553.

[43] Kadonaga J T. Regulation of RNA polymerase II transcription by sequence-specific DNA binding factors. Cell,2004,116(2):247-257.

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