Hypoxia induces epithelial-mesenchymal transition via activation of SNAI1 by hypoxia-inducible factor -1α in hepatocellular carcinoma

BMC Cancer

Lin Zhang³  Gang Huang¹  Xiaowu Li⁵  Yujun Zhang⁵  Yan Jiang⁵  Junjie Shen⁴  Jia Liu⁴  Qingliang Wang⁴  Jin Zhu⁵  Xiaobin Feng⁵  Jiahong Dong²  Cheng Qian⁴ 

[1] Department of Medical Genetics, College of Basic Medicine, Third Military Medical University, Chongqing, China

[2] Institute of Hepatobiliary Surgery, General Hospital of PLA, Beijing, 100853, PR China

[3] Department of Radiology, Southwest Hospital, Third Military Medical University, Chongqing, China

[4] Institute of Pathology and Southwest Cancer Center, Southwest Hospital, Third Military Medical University, Chongqing, China

[5] Institute of Hepatobiliary Surgery, Southwest Hospital, Third Military Medical University, Chongqing, China

关键词: Hepatocellular carcinoma;    SNAI1;    Hypoxia-inducible factor-1α;    Hypoxia;    Epithelial-mesenchymal transition;   

Others  :  1079882
DOI  :  10.1186/1471-2407-13-108

 received in 2012-09-20, accepted in 2013-02-25,  发布年份 2013

【 摘 要 】

Background

High invasion and metastasis are the primary factors causing poor prognosis of patients with hepatocellular carcinoma (HCC). However, the molecular mechanisms underlying these biological behaviors have not been completely elucidated. In this study, we investigate the molecular mechanism by which hypoxia promotes HCC invasion and metastasis through inducing epithelial-mesenchymal transition (EMT).

Methods

The expression of EMT markers was analyzed by immunohistochemistry. Effect of hypoxia on induction of EMT and ability of cell migration and invasion were performed. Luciferase reporter system was used for evaluation of Snail regulation by hypoxia-inducible factor -1α (HIF-1α).

Results

We found that overexpression of HIF-1α was observed in HCC liver tissues and was related to poor prognosis of HCC patients. HIF-1α expression profile was correlated with the expression levels of SNAI1, E-cadherin, N-cadherin and Vimentin. Hypoxia was able to induce EMT and enhance ability of invasion and migration in HCC cells. The same phenomena were also observed in CoCl2-treated cells. The shRNA-mediated HIF-1α suppression abrogated CoCl2-induced EMT and reduced ability of migration and invasion in HCC cells. Luciferase assay showed that HIF-1α transcriptional regulated the expression of SNAI1 based on two hypoxia response elements (HREs) in SNAI1 promoter.

Conclusions

We demonstrated that hypoxia-stabilized HIF1α promoted EMT through increasing SNAI1 transcription in HCC cells. This data provided a potential therapeutic target for HCC treatment.

【 授权许可】

   
2013 Zhang et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Christofori G: New signals from the invasive front. Nature 2006, 441:444-450.
• [2]Yang MH, Wu MZ, Chiou SH, Chen PM, Chang SY, Liu CJ, Teng SC, Wu KJ: Direct regulation of TWIST by HIF-1alpha promotes metastasis. Nat Cell Biol 2008, 10:295-305.
• [3]Thompson EW, Williams ED: EMT and MET in carcinoma–clinical observations, regulatory pathways and new models. Clin Exp Metastasis 2008, 25:591-592.
• [4]Hill RP, Marie-Egyptienne DT, Hedley DW: Cancer stem cells, hypoxia and metastasis. Semin Radiat Oncol 2009, 19:106-111.
• [5]Kudo-Saito C, Shirako H, Takeuchi T, Kawakami Y: Cancer Metastasis Is Accelerated through Immunosuppression during Snail-induced EMT of Cancer Cells. Cancer Cell 2009, 15:195-206.
• [6]Yang MH, Wu KJ: TWIST activation by hypoxia inducible factor-1 (HIF-1): implications in metastasis and development. Cell Cycle 2008, 7:2090-2096.
• [7]Wang Y, Xue TC, Xie XY, Chen Y, Ye SL, Ren ZG: [Relationship between epithelial-mesenchymal transition and lung metastasis in hepatocellular carcinoma]. Zhonghua Wai Ke Za Zhi 2008, 46:1624-1627.
• [8]Brabletz T, Hlubek F, Spaderna S, Schmalhofer O, Hiendlmeyer E, Jung A, Kirchner T: Invasion and metastasis in colorectal cancer: epithelial-mesenchymal transition, mesenchymal-epithelial transition, stem cells and beta-catenin. Cells Tissues Organs 2005, 179:56-65.
• [9]Wu Y, Zhou BP: Inflammation: A driving force speeds cancer metastasis. Cell Cycle 2009, 8:3267-3273.
• [10]Molloy T, van 't Veer LJ: Recent advances in metastasis research. Curr Opin Genet Dev 2008, 18:35-41.
• [11]Robinson BD, Sica GL, Liu YF, Rohan TE, Gertler FB, Condeelis JS, Jones JG: Tumor microenvironment of metastasis in human breast carcinoma: a potential prognostic marker linked to hematogenous dissemination. Clin Cancer Res 2009, 15:2433-2441.
• [12]Mendoza M, Khanna C: Revisiting the seed and soil in cancer metastasis. Int J Biochem Cell Biol 2009, 41:1452-1462.
• [13]Melnikova VO, Bar-Eli M: Inflammation and melanoma metastasis. Pigment Cell Melanoma Res 2009, 22:257-267.
• [14]Erler JT, Weaver VM: Three-dimensional context regulation of metastasis. Clin Exp Metastasis 2009, 26:35-49.
• [15]Chambers AF: Influence of diet on metastasis and tumor dormancy. Clin Exp Metastasis 2009, 26:61-66.
• [16]Wikman H, Vessella R, Pantel K: Cancer micrometastasis and tumour dormancy. APMIS 2008, 116:754-770.
• [17]Taylor J, Hickson J, Lotan T, Yamada DS, Rinker-Schaeffer C: Using metastasis suppressor proteins to dissect interactions among cancer cells and their microenvironment. Cancer Metastasis Rev 2008, 27:67-73.
• [18]Tse JC, Kalluri R: Mechanisms of metastasis: epithelial-to-mesenchymal transition and contribution of tumor microenvironment. J Cell Biochem 2007, 101:816-829.
• [19]Lorusso G, Ruegg C: The tumor microenvironment and its contribution to tumor evolution toward metastasis. Histochem Cell Biol 2008, 130:1091-1103.
• [20]Gout S, Huot J: Role of cancer microenvironment in metastasis: focus on colon cancer. Cancer Microenviron 2008, 1:69-83.
• [21]Blouin S, Basle MF, Chappard D: Interactions between microenvironment and cancer cells in two animal models of bone metastasis. Br J Cancer 2008, 98:809-815.
• [22]Bidard FC, Pierga JY, Vincent-Salomon A, Poupon MF: A "class action" against the microenvironment: do cancer cells cooperate in metastasis? Cancer Metastasis Rev 2008, 27:5-10.
• [23]Albini A, Mirisola V, Pfeffer U: Metastasis signatures: genes regulating tumor-microenvironment interactions predict metastatic behavior. Cancer Metastasis Rev 2008, 27:75-83.
• [24]Harrison L, Blackwell K: Hypoxia and anemia: factors in decreased sensitivity to radiation therapy and chemotherapy? Oncologist 2004, 9:31-40.
• [25]Chaudary N, Hill RP: Increased expression of metastasis-related genes in hypoxic cells sorted from cervical and lymph nodal xenograft tumors. Lab Invest 2009, 89:587-596.
• [26]Hu XB, Feng F, Wang YC, Wang L, He F, Dou GR, Liang L, Zhang HW, Liang YM, Han H: Blockade of Notch signaling in tumor-bearing mice may lead to tumor regression, progression, or metastasis, depending on tumor cell types. Neoplasia 2009, 11:32-38.
• [27]Wu H, Liang X, Fang Y, Qin X, Zhang Y, Liu J: Resveratrol inhibits hypoxia-induced metastasis potential enhancement by restricting hypoxia-induced factor-1 alpha expression in colon carcinoma cells. Biomed Pharmacother 2008, 62:613-621.
• [28]Semenza GL: Hydroxylation of HIF-1: oxygen sensing at the molecular level. Physiology (Bethesda) 2004, 19:176-182.
• [29]Miyake K, Yoshizumi T, Imura S, Sugimoto K, Batmunkh E, Kanemura H, Morine Y, Shimada M: Expression of hypoxia-inducible factor-1alpha, histone deacetylase 1, and metastasis-associated protein 1 in pancreatic carcinoma: correlation with poor prognosis with possible regulation. Pancreas 2008, 36:e1-e9.
• [30]Liao D, Corle C, Seagroves TN, Johnson RS: Hypoxia-inducible factor-1 alpha is a key regulator of metastasis in a transgenic model of cancer initiation and progression. Cancer Res 2007, 67:563-572.
• [31]Chen HHW, Su WC, Lin PW, Guo HR, Lee WY: Hypoxia-inducible factor-1 alpha correlates with MET and metastasis in node-negative breast cancer. Breast Cancer Res Treat 2007, 103:167-175.
• [32]Barnhart BC, Simon MC: Metastasis and stem cell pathways. Cancer Metastasis Rev 2007, 26:261-271.
• [33]Schindl M, Schoppmann SF, Samonigg H, Hausmaninger H, Kwasny W, Gnant M, Jakesz R, Kubista E, Birner P, Oberhuber G: Overexpression of hypoxia-inducible factor 1alpha is associated with an unfavorable prognosis in lymph node-positive breast cancer. Clin Cancer Res 2002, 8:1831-1837.
• [34]Bos R, van der Groep P, Greijer AE, Shvarts A, Meijer S, Pinedo HM, Semenza GL, van Diest PJ, van der Wall E: Levels of hypoxia-inducible factor-1alpha independently predict prognosis in patients with lymph node negative breast carcinoma. Cancer 2003, 97:1573-1581.
• [35]Hanahan D, Weinberg RA: The hallmarks of cancer. Cell 2000, 100:57-70.
• [36]Evans AJ, Russell RC, Roche O, Burry TN, Fish JE, Chow VW, Kim WY, Saravanan A, Maynard MA, Gervais ML: VHL promotes E2 box-dependent E-cadherin transcription by HIF-mediated regulation of SIP1 and snail. Mol Cell Biol 2007, 27:157-169.
• [37]Chen HH, Su WC, Lin PW, Guo HR, Lee WY: Hypoxia-inducible factor-1alpha correlates with MET and metastasis in node-negative breast cancer. Breast Cancer Res Treat 2007, 103:167-175.
• [38]Postovit LM, Abbott DE, Payne SL, Wheaton WW, Margaryan NV, Sullivan R, Jansen MK, Csiszar K, Hendrix MJ, Kirschmann DA: Hypoxia/reoxygenation: a dynamic regulator of lysyl oxidase-facilitated breast cancer migration. J Cell Biochem 2008, 103:1369-1378.
• [39]Haase VH: Oxygen regulates epithelial-to-mesenchymal transition: insights into molecular mechanisms and relevance to disease. Kidney Int 2009, 76:492-499.
• [40]Huang K, Sneed PK, Kunwar S, Kragten A, Larson DA, Berger MS, Chan A, Pouliot J, McDermott MW: Surgical resection and permanent iodine-125 brachytherapy for brain metastases. J Neurooncol 2009, 91:83-93.
• [41]Pignol JP, Rakovitch E, Keller BM, Sankreacha R, Chartier C: Tolerance and acceptance results of a palladium-103 permanent breast seed implant Phase I/II study. Int J Radiat Oncol Biol Phys 2009, 73:1482-1488.
• [42]Roodink I, van der Laak J, Kusters B, Wesseling P, Verrijp K, de Waal R, Leenders W: Development of the tumor vascular bed in response to hypoxia-induced VEGF-A differs from that in tumors with constitutive VEGF-A expression. Int J Cancer 2006, 119:2054-2062.
• [43]Staller P, Sulitkova J, Lisztwan J, Moch H, Oakeley EJ, Krek W: Chemokine receptor CXCR4 downregulated by von Hippel-Lindau tumour suppressor pVHL. Nature 2003, 425:307-311.
• [44]Semenza GL, Agani F, Feldser D, Iyer N, Kotch L, Laughner E, Yu A: Hypoxia, HIF-1, and the pathophysiology of common human diseases. Adv Exp Med Biol 2000, 475:123-130.
• [45]Erler JT, Bennewith KL, Nicolau M, Dornhofer N, Kong C, Le QT, Chi JT, Jeffrey SS, Giaccia AJ: Lysyl oxidase is essential for hypoxia-induced metastasis. Nature 2006, 440:1222-1226.
• [46]Boyer B, Thiery JP: Epithelium-mesenchyme interconversion as example of epithelial plasticity. Apmis 1993, 101:257-268.
• [47]Zavadil J, Bitzer M, Liang D, Yang YC, Massimi A, Kneitz S, Piek E, Bottinger EP: Genetic programs of epithelial cell plasticity directed by transforming growth factor-beta. Proc Natl Acad Sci U S A 2001, 98:6686-6691.
• [48]Janda E, Lehmann K, Killisch I, Jechlinger M, Herzig M, Downward J, Beug H, Grunert S: Ras and TGF[beta] cooperatively regulate epithelial cell plasticity and metastasis: dissection of Ras signaling pathways. J Cell Biol 2002, 156:299-313.
• [49]He X, Liu J, Yang C, Su C, Zhou C, Zhang Q, Li L, Wu H, Liu X, Wu M, Qian Q: 5/35 fiber-modified conditionally replicative adenovirus armed with p53 shows increased tumor-suppressing capacity to breast cancer cells. Hum Gene Ther 2011, 22:283-292.
• [50]Yang MH, Chen CL, Chau GY, Chiou SH, Su CW, Chou TY, Peng WL, Wu JC: Comprehensive analysis of the independent effect of twist and snail in promoting metastasis of hepatocellular carcinoma. Hepatology 2009, 50:1464-1474.
• [51]Dang H, Ding W, Emerson D, Rountree CB: Snail1 induces epithelial-to-mesenchymal transition and tumor initiating stem cell characteristics. BMC Cancer 2011, 11:396. BioMed Central Full Text
• [52]Copple BL: Hypoxia stimulates hepatocyte epithelial to mesenchymal transition by hypoxia-inducible factor and transforming growth factor-beta-dependent mechanisms. Liver Int 2010, 30:669-682.
• [53]Luo D, Wang J, Li J, Post M: Mouse snail is a target gene for HIF. Mol Cancer Res 2011, 9:234-245.
• [54]Liu S, Kumar SM, Martin JS, Yang R, Xu X: Snail1 mediates hypoxia-induced melanoma progression. Am J Pathol 2011, 179:3020-3031.