Overexpression of brachyury contributes to tumor metastasis by inducing epithelial-mesenchymal transition in hepatocellular carcinoma

Journal of Experimental & Clinical Cancer Research

Jingbo Kang¹  Sudong Wu¹  Henghu Fang¹  Xiaoning Lv²  Shanshan Wu¹  Rui Du¹ 

[1] Department of Radiation Oncology and Integrative Oncology, Navy General Hospital, 6# Fu Cheng Road, Beijing 100037, People’s Republic of China;Department of Aviation and Diving, Navy General Hospital, Beijing, People’s Republic of China

关键词: PTEN/Akt/Snail-Dependent Pathway;    Epithelial-Mesenchymal Transition;    Cancer metastasis;    Brachyury;    Hepatocellular carcinoma;   

Others  :  1145481
DOI  :  10.1186/s13046-014-0105-6

 received in 2014-08-12, accepted in 2014-11-24,  发布年份 2014

【 摘 要 】

Aims

Brachyury overexpression has been reported in various human malignant neoplasms, but its expression and function in hepatocellular carcinoma progression and metastasis remains unknown. The present study aimed to evaluate the critical role of Brachyury in HCC metastasis.

Methods

The expression of Brachyury in human HCC (SMMC7721, HepG2, FHCC98, and Hep3B) and control cell lines was analyzed using quantitative reverse-transcriptase polymerase chain reaction and immunoflourence methods. Cancerous tissues collected from patients with HCC (n = 112) were analyzed using immunohistochemical method; a microarray analysis of HCC tissues was performed to explore the clinicopathological variables of HCC. The migratory and invasive capacities of Brachyury-SMMC7721 and Brachyury-HepG2 transfected cells were evaluated using in vitro scratch wound healing and Matrigel invasion assays, respectively. Further, six-week-old male BALB/c nude mice (n = 10) model was used in vivo assay.

Results

Elevated expression of Brachyury was detected in HCCs (62.5%) compared with that in adjacent nontumorous tissues. Clinicopathological analysis revealed a close correlation of Brachyury expression with distant metastasis and poor prognosis of HCC. Overexpression of Brachyury promoted epithelial-mesenchymal transition (EMT) and metastasis of HCC cells in vitro and in vivo. Brachyury overexpression enhanced Akt activation by inhibiting phosphatase and tensin homolog (PTEN), which led to subsequent stabilization of Snail, a critical EMT mediator.

Conclusion

The study findings suggest that elevated Brachyury facilitates HCC metastasis by promoting EMT via PTEN/Akt/Snail-dependent pathway. Brachyury plays a pivotal role in HCC metastasis and may serve as a novel prognostic biomarker and therapeutic target.

【 授权许可】

   
2014 Du et al.; licensee BioMed Central.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Farazi PA, DePinho RA: Hepatocellular carcinoma pathogenesis: from genes to environment. Nat Rev Cancer 2006, 6:674-687.
• [2]Mathurin P, Rixe O, Carbonell N, Bernard B, Cluzel P, Bellin MF, Khayat D, Opolon P, Poynard T: Review article: overview of medical treatments in unresectable hepatocellular carcinoma-an impossible meta-analysis? Aliment Pharmacol Ther 1998, 12:111-126.
• [3]Kawano Y, Sasaki A, Kai S, Endo Y, Iwaki K, Uchida H, Shibata K, Ohta M, Kitano S: Prognosis of patients with intrahepatic recurrence after hepatic resection for hepatocellular carcinoma: a retrospective study. European J Surg Oncol J Eur Soc Surg Oncol Br Assoc Surg Oncol 2009, 35:174-179.
• [4]Liotta LA: Mechanisms of cancer invasion and metastasis.Important advances in oncology 1985:28–41.
• [5]Thiery JP, Acloque H, Huang RY, Nieto MA: Epithelial-mesenchymal transitions in development and disease. Cell 2009, 139:871-890.
• [6]Larue L, Bellacosa A: Epithelial-mesenchymal transition in development and cancer: role of phosphatidylinositol 3' kinase/AKT pathways. Oncogene 2005, 24:7443-7454.
• [7]Christofori G: New signals from the invasive front. Nature 2006, 441:444-450.
• [8]Boivin D, Bilodeau D, Beliveau R: Regulation of cytoskeletal functions by Rho small GTP-binding proteins in normal and cancer cells. Can J Physiol Pharmacol 1996, 74:801-810.
• [9]Niu RF, Zhang L, Xi GM, Wei XY, Yang Y, Shi YR, Hao XS: Up-regulation of Twist induces angiogenesis and correlates with metastasis in hepatocellular carcinoma. J Exp Clin Cancer Res CR 2007, 26:385-394.
• [10]Teng Y, Zeisberg M, Kalluri R: Transcriptional regulation of epithelial-mesenchymal transition. J Clin Invest 2007, 117:304-306.
• [11]Naiche LA, Harrelson Z, Kelly RG, Papaioannou VE: T-box genes in vertebrate development. Annu Rev Genet 2005, 39:219-239.
• [12]Zhou B, Chen H, Wei D, Kuang Y, Zhao X, Li G, Xie J, Chen P: A novel miR-219-SMC4-JAK2/Stat3 regulatory pathway in human hepatocellular carcinoma. J Exp Clin Cancer Res 2014, 33:55. BioMed Central Full Text
• [13]Beddington RS, Rashbass P, Wilson V: Brachyury-a gene affecting mouse gastrulation and early organogenesis.Dev Suppl 1992, 157-165.
• [14]Schulte-Merker S, van Eeden FJ, Halpern ME, Kimmel CB, Nusslein-Volhard C: no tail (ntl) is the zebrafish homologue of the mouse T (Brachyury) gene. Development 1994, 120:1009-1015.
• [15]Wilson V, Manson L, Skarnes WC, Beddington RS: The T gene is necessary for normal mesodermal morphogenetic cell movements during gastrulation. Development 1995, 121:877-886.
• [16]Showell C, Binder O, Conlon FL: T-box genes in early embryogenesis. Dev Dyn Off Publ Am Assoc Anat 2004, 229:201-218.
• [17]Griffin KJP, Kimelman D: One-Eyed Pinhead and Spadetail are essential for heart and somite formation. Nat Cell Biol 2002, 4:821-825.
• [18]Kilic N, Feldhaus S, Kilic E, Tennstedt P, Wicklein D, Wasielewski R, Viebahn C, Kreipe H, Schumacher U: Brachyury expression predicts poor prognosis at early stages of colorectal cancer. Eur J Cancer 2011, 47:1080-1085.
• [19]Yang XR, Ng D, Alcorta DA, Liebsch NJ, Sheridan E, Li S, Goldstein AM, Parry DM, Kelley MJ: T (Brachyury) gene duplication confers major susceptibility to familial chordoma. Nat Genet 2009, 41:1176-1178.
• [20]Fernando RI, Litzinger M, Trono P, Hamilton DH, Schlom J, Palena C: The T-box transcription factor Brachyury promotes epithelial-mesenchymal transition in human tumor cells. J Clin Invest 2010, 120:533-544.
• [21]Huang B, Cohen JR, Fernando RI, Hamilton DH, Litzinger MT, Hodge JW, Palena C: The embryonic transcription factor Brachyury blocks cell cycle progression and mediates tumor resistance to conventional antitumor therapies. Cell Death Dis 2013, 4:e682.
• [22]Imajyo I, Sugiura T, Kobayashi Y, Shimoda M, Ishii K, Akimoto N, Yoshihama N, Kobayashi I, Mori Y: T-box transcription factor Brachyury expression is correlated with epithelial-mesenchymal transition and lymph node metastasis in oral squamous cell carcinoma. Int J Oncol 2012, 41:1985-1995.
• [23]Wong N, Lai P, Lee SW, Fan S, Pang E, Liew CT, Sheng Z, Lau JW, Johnson PJ: Assessment of genetic changes in hepatocellular carcinoma by comparative genomic hybridization analysis: relationship to disease stage, tumor size, and cirrhosis. Am J Pathol 1999, 154:37-43.
• [24]Weiss-Steider B, Soto-Cruz I, Martinez-Campos CA, Mendoza-Rincon JF: Expression of MICA, MICB and NKG2D in human leukemic myelomonocytic and cervical cancer cells. J Exp Clin Cancer Res 2011, 30:37. BioMed Central Full Text
• [25]Chen L, Chan TH, Yuan YF, Hu L, Huang J, Ma S, Wang J, Dong SS, Tang KH, Xie D, Li Y, Guan XY: CHD1L promotes hepatocellular carcinoma progression and metastasis in mice and is associated with these processes in human patients. CHD1L promotes hepatocellular carcinoma progression and metastasis in mice and is associated with these processes in human patients. J Clin Invest 2010, 120:1178-1191.
• [26]Thiery JP, Sleeman JP: Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol 2006, 7:131-142.
• [27]Ning J, Liu W, Zhang J, Lang Y, Xu S: Ran GTPase Induces EMT and Enhances Invasion in Non-Small Cell Lung Cancer Cells Through Activation of PI3K-AKT Pathway. Oncol Res Featuring Preclin Clin Cancer Ther 2014, 21:67-72.
• [28]Ginnebaugh KR, Ahmad A, Sarkar FH: The therapeutic potential of targeting the epithelial-mesenchymal transition in cancer. Expert Opin Ther Targets 2014, 18:731-745.
• [29]Tian H, Ge C, Li H, Zhao F, Hou H, Chen T, Jiang G, Xie H, Cui Y, Yao M, Li J: Ribonucleotide reductase M2B inhibits cell migration and spreading by early growth response protein 1-mediated phosphatase and tensin homolog/Akt1 pathway in hepatocellular carcinoma. Hepatology 2014, 59:1459-1470.
• [30]Chen KC, Chen CY, Lin CJ, Yang TY, Chen TH, Wu LC, Wu CC: Luteolin attenuates TGF-beta1-induced epithelial-mesenchymal transition of lung cancer cells by interfering in the PI3K/Akt-NF-kappaB-Snail pathway. Life Sci 2013, 93:924-933.
• [31]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.
• [32]Roselli M, Fernando RI, Guadagni F, Spila A, Alessandroni J, Palmirotta R, Costarelli L, Litzinger M, Hamilton D, Huang B, Tucker J, Tsang KY, Schlom J, Palena C: Brachyury, a driver of the epithelial-mesenchymal transition, is overexpressed in human lung tumors: an opportunity for novel interventions against lung cancer. Clin Cancer Res 2012, 18:3868-3879.
• [33]Haro A, Yano T, Kohno M, Yoshida T, Koga T, Okamoto T, Takenoyama M, Maehara Y: Expression of Brachyury gene is a significant prognostic factor for primary lung carcinoma. Ann Surg Oncol 2013, 20(Suppl 3):S509-S516.
• [34]Palena C, Roselli M, Litzinger MT, Ferroni P, Costarelli L, Spila A, Cavaliere F, Huang B, Fernando RI, Hamilton DH, Jochems C, Tsang KY, Cheng Q, Kim Lyerly H, Schlom J, Guadagni F: Overexpression of the EMT driver brachyury in breast carcinomas: association with poor prognosis. J Natl Cancer Inst 2014, 9:106(5).
• [35]Pinto F, Pértega-Gomes N, Pereira MS, Vizcaíno JR, Monteiro P, Henrique RM, Baltazar F, Andrade RP, Reis RM: T-box transcription factor brachyury is associated with prostate cancer progression and aggressiveness. Clin Cancer Res 2014, 20:4949-4961.
• [36]Lim J, Thiery JP: Epithelial-mesenchymal transitions: insights from development. Development 2012, 139:3471-3486.
• [37]Thiery JP: Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer 2002, 2:442-454.
• [38]Larocca C, Cohen JR, Fernando RI, Huang B, Hamilton DH, Palena C: An Autocrine Loop between TGF-β1 and the Transcription Factor Brachyury Controls the Transition of Human Carcinoma Cells into a Mesenchymal Phenotype. Mol Cancer Ther 2013, 12:1805-1815.
• [39]Leslie NR, Yang X, Downes CP, Weijer CJ: PtdIns(3,4,5)P3-Dependent and -Independent Roles for PTEN in the Control of Cell Migration. Curr Biol 2007, 17:115-125.