Characterization of genome-wide TFCP2 targets in hepatocellular carcinoma: implication of targets FN1 and TJP1 in metastasis

Journal of Experimental & Clinical Cancer Research

Shusen Zheng²  Xuyong Wei¹  Haijun Guo²  Jianguo Wang¹  Qi Ling²  Jun Cheng²  Haiyang Xie¹  Lin Zhou¹  Zhikun Liu²  Xiao Xu² 

[1] Key Lab of Combined Multi-Organ Transplantation, Ministry of Public Health, First Affiliated Hospital, Zhejiang University School of Medicine, HangZhou, China;Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, Zhejiang University School of Medicine, 79 QingChun Road, HangZhou, China

关键词: Transcription factor CP2;    Tight junction protein 1;    Fibronectin 1;    Metastasis;    Hepatocellular carcinoma;   

Others  :  1133645
DOI  :  10.1186/s13046-015-0121-1

 received in 2014-09-05, accepted in 2015-01-04,  发布年份 2015

【 摘 要 】

Background

Transcription factor CP2 (TFCP2) is overexpressed in hepatocellular carcinoma(HCC) and correlated with the progression of the disease. Here we report the use of an integrated systems biology approach to identify genome-wide scale map of TFCP2 targets as well as the molecular function and pathways regulated by TFCP2 in HCC.

Methods

We combined Chromatin immunoprecipitation (ChIP) on chip along with gene expression microarrays to study global transcriptional regulation of TFCP2 in HCC. The biological functions, molecular pathways, and networks associated with TFCP2 were identified using computational approaches. Validation of selected target gene expression and direct binding of TFCP2 to promoters were performed by ChIP -PCR and promoter reporter.

Results

TFCP2 fostered a highly aggressive and metastatic phenotype in different HCC cells. Transcriptome analysis showed that alteration of TFCP2 in HCC cells led to change of genes in biological functions involved in cancer, cellular growth and proliferation, angiogenesis, cell movement and attachment. Pathways related to cell movement and cancer progression were also enriched. A quest for TFCP2-regulated factors contributing to metastasis, by integration of transcriptome and ChIP on chip assay, identified fibronectin 1 (FN1) and tight junction protein 1 (TJP1) as targets of TFCP2, and as key mediators of HCC metastasis. Promoter reporter identified the TFCP2-responsive region, and located the motifs of TFCP2-binding sites in the FN1 promoter, which then was confirmed by ChIP-PCR. We further showed that FN1 inhibition blocks the TFCP2-induced increase in HCC cell aggression, and that overexpression of TFCP2 can rescue the effects of FN1 inhibition. Knock down of TJP1 could also rescue, at least in part, the aggressive effect of TFCP2 knockdown in HCC cells.

Conclusions

The identification of global targets, molecular pathways and networks associated with TFCP2, together with the discovery of the effect of TFCP2 on FN1 and TJP1 that are involved in metastasis, adds to our understanding of the mechanisms that determine a highly aggressive and metastatic phenotype in hepatocarcinogenesis.

【 授权许可】

   
2015 Xu et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Bruix J, Sherman M: Management of hepatocellular carcinoma: an update. Hepatology 2011, 53(3):1020-2. doi:10.1002/hep.24199
• [2]Miki D, Ochi H, Hayes CN, Aikata H, Chayama K: Hepatocellular carcinoma: towards personalized medicine. Cancer Sci 2012, 103(5):846-50. doi:10.1111/j.1349-7006.2012.02242.x
• [3]Schutte K, Bornschein J, Malfertheiner P: Hepatocellular carcinoma–epidemiological trends and risk factors. Dig Dis 2009, 27(2):80-92. doi:10.1159/000218339
• [4]Trinchet JC, Nahon P, Ganne-Carrie N: [Screening of hepatocellular carcinoma and surveillance of patients with cirrhosis: a public health issue]. Rev Prat 2013, 63(2):222-5.
• [5]Kaufhold S, Bonavida B: Central role of Snail1 in the regulation of EMT and resistance in cancer: a target for therapeutic intervention. J Exp Clin Cancer Res 2014, 33:62. doi:10.1186/s13046-014-0062-0 BioMed Central Full Text
• [6]Dong S, Kong J, Kong F, Gao J, Ke S, Wang S, et al.: Insufficient radiofrequency ablation promotes epithelial-mesenchymal transition of hepatocellular carcinoma cells through Akt and ERK signaling pathways. J Transl Med. 2013, 11:273. doi:10.1186/1479-5876-11-273 BioMed Central Full Text
• [7]Kachroo P, Lee MH, Zhang L, Baratelli F, Lee G, Srivastava MK, et al.: IL-27 inhibits epithelial-mesenchymal transition and angiogenic factor production in a STAT1-dominant pathway in human non-small cell lung cancer. J Exp Clin Cancer Res 2013, 32:97. doi:10.1186/1756-9966-32-97 BioMed Central Full Text
• [8]Zhu L, Qin H, Li PY, Xu SN, Pang HF, Zhao HZ, et al.: Response gene to complement-32 enhances metastatic phenotype by mediating transforming growth factor beta-induced epithelial-mesenchymal transition in human pancreatic cancer cell line BxPC-3. J Exp Clin Cancer Res 2012, 31:29. doi:10.1186/1756-9966-31-29 BioMed Central Full Text
• [9]Kim CH, Heath C, Bertuch A, Hansen U: Specific stimulation of simian virus 40 late transcription in vitro by a cellular factor binding the simian virus 40 21-base-pair repeat promoter element. Proc Natl Acad Sci U S A 1987, 84(17):6025-9.
• [10]Veljkovic J, Hansen U: Lineage-specific and ubiquitous biological roles of the mammalian transcription factor LSF. Gene 2004, 343(1):23-40. doi:10.1016/j.gene.2004.08.010
• [11]Hansen U, Owens L, Saxena UH: Transcription factors LSF and E2Fs: tandem cyclists driving G0 to S? Cell Cycle 2009, 8(14):2146-51.
• [12]Saxena UH, Powell CM, Fecko JK, Cacioppo R, Chou HS, Cooper GM, et al.: Phosphorylation by cyclin C/cyclin-dependent kinase 2 following mitogenic stimulation of murine fibroblasts inhibits transcriptional activity of LSF during G1 progression. Mol Cell Biol 2009, 29(9):2335-45. doi:10.1128/mcb. 00687-08
• [13]Porta-de-la-Riva M, Stanisavljevic J, Curto J, Franci C, Diaz VM, Garcia de Herreros A, et al.: TFCP2c/LSF/LBP-1c is required for Snail1-induced fibronectin gene expression. Biochem J 2011, 435(3):563-8. doi:10.1042/bj20102057
• [14]Yoo BK, Emdad L, Gredler R, Fuller C, Dumur CI, Jones KH, et al.: Transcription factor Late SV40 Factor (LSF) functions as an oncogene in hepatocellular carcinoma. Proc Natl Acad Sci U S A 2010, 107(18):8357-62. doi:10.1073/pnas.1000374107
• [15]Santhekadur PK, Gredler R, Chen D, Siddiq A, Shen XN, Das SK, et al.: Late SV40 factor (LSF) enhances angiogenesis by transcriptionally up-regulating matrix metalloproteinase-9 (MMP-9). J Biol Chem 2012, 287(5):3425-32. doi:10.1074/jbc.M111.298976
• [16]Powell CM, Rudge TL, Zhu Q, Johnson LF, Hansen U: Inhibition of the mammalian transcription factor LSF induces S-phase-dependent apoptosis by downregulating thymidylate synthase expression. EMBO J 2000, 19(17):4665-75. doi:10.1093/emboj/19.17.4665
• [17]Yoo BK, Gredler R, Vozhilla N, Su ZZ, Chen D, Forcier T, et al.: Identification of genes conferring resistance to 5-fluorouracil. Proc Natl Acad Sci U S A 2009, 106(31):12938-43. doi:10.1073/pnas.0901451106
• [18]Cheng J, Xie HY, Xu X, Wu J, Wei X, Su R, et al.: NDRG1 as a biomarker for metastasis, recurrence and of poor prognosis in hepatocellular carcinoma. Cancer Lett 2011, 310(1):35-45. doi:10.1016/j.canlet.2011.06.001
• [19]Xu X, Wei X, Ling Q, Cheng J, Zhou B, Xie H, et al.: Identification of two portal vein tumor thrombosis associated proteins in hepatocellular carcinoma: protein disulfide-isomerase A6 and apolipoprotein A-I. J Gastroenterol Hepatol 2011, 26(12):1787-94. doi:10.1111/j.1440-1746.2011.06796.x
• [20]Wang F, Song G, Liu M, Li X, Tang H: miRNA-1 targets fibronectin1 and suppresses the migration and invasion of the HEp2 laryngeal squamous carcinoma cell line. FEBS Lett 2011, 585(20):3263-9. doi:10.1016/j.febslet.2011.08.052
• [21]Qian P, Zuo Z, Wu Z, Meng X, Li G, Zhang W, et al.: Pivotal role of reduced let-7 g expression in breast cancer invasion and metastasis. Cancer Res 2011, 71(20):6463-74. doi:10.1158/0008-5472.can-11-1322
• [22]Murata T, Nitta M, Yasuda K: Transcription factor CP2 is essential for lens-specific expression of the chicken alphaA-crystallin gene. Genes Cells 1998, 3(7):443-57.
• [23]Chae JH, Lee YH, Kim CG: Transcription factor CP2 is crucial in hemoglobin synthesis during erythroid terminal differentiation in vitro. Biochem Biophys Res Commun 1999, 263(2):580-3. doi:10.1006/bbrc.1999.1408
• [24]Yoo BK, Gredler R, Chen D, Santhekadur PK, Fisher PB, Sarkar D: c-Met activation through a novel pathway involving osteopontin mediates oncogenesis by the transcription factor LSF. J Hepatol 2011, 55(6):1317-24. doi:10.1016/j.jhep.2011.02.036
• [25]Chen J, Wang T, Zhou YC, Gao F, Zhang ZH, Xu H, et al.: Aquaporin 3 promotes epithelial-mesenchymal transition in gastric cancer. J Exp Clin Cancer Res 2014, 33:38. doi:10.1186/1756-9966-33-38 BioMed Central Full Text
• [26]Lida R, Yasuda T, Kishi K: Identification of novel fibronectin fragments detected specifically in juvenile urine. Febs J 2007, 274(15):3939-47. doi:10.1111/j.1742-4658.2007.05926.x
• [27]Meier C, Spitschak A, Abshagen K, Gupta S, Mor JM, Wolkenhauer O, et al.: Association of RHAMM with E2F1 promotes tumour cell extravasation by transcriptional up-regulation of fibronectin. J Pathol 2014, 234(3):351-64.
• [28]Howe EN, Cochrane DR, Richer JK: Targets of miR-200c mediate suppression of cell motility and anoikis resistance. Breast Cancer Res 2011, 13(2):R45. doi:10.1186/bcr2867 BioMed Central Full Text