The role of TGFBI in mesothelioma and breast cancer: association with tumor suppression

BMC Cancer

Bingyan Li²  Gengyun Wen²  Yongliang Zhao²  Jian Tong³  Tom K Hei¹ 

[1] Department of Radiation Oncology, College of Physicians and Surgeons, Columbia University, 630 West 168th St, New York, NY, 10032, USA

[2] Center for Radiological Research, College of Physicians & Surgeons, Columbia University, New York, NY, USA

[3] School of Radiation Medicine and Public Health, Soochow University, Suzhou, China

关键词: Proliferation;    Breast tumor;    Mesothelioma;    Tumor suppressor;    TGFBI;   

Others  :  1080368
DOI  :  10.1186/1471-2407-12-239

 received in 2011-11-13, accepted in 2012-05-21,  发布年份 2012

【 摘 要 】

Background

Transforming growth factor β induced (TGFBI) product, an extracellular matrix (ECM) protein, has been implicated as a putative tumor suppressor in recent studies. Our previous findings revealed that expression of TGFBI gene is down-regulated in a variety of cancer cell lines and clinical tissue samples. In this study, ectopic expression of TGFBI was used to ascertain its role as a tumor suppressor and to determine the underlying mechanism of mesothelioma and breast cancer.

Methods

Cells were stably transfected with pRc/CMV2-TGFBI and pRc/CMV2-empty vector with Lipofectamine Plus. Ectopic expression of TGFBI was quantified by using quantitative PCR and Western-blotting. Characterization of cell viability was assessed using growth curve, clonogenic survival and soft agar growth. The potential of tumor formation was evaluated by an in vivo mouse model. Cell cycle was analyzed via flow cytometry. Expressions of p21, p53, p16 and p14 were examined using Western-blotting. Senescent cells were sorted by using a Senescence β-Galactosidase Staining Kit. Telomerase activity was measured using quantitative telomerase detection kit.

Results

In this study, an ectopic expression of TGFBI in two types of cancer cell lines, a mesothelioma cell line NCI-H28 and a breast cancer cell line MDA-MB-231 was found to have reduced the cellular growth, plating efficiency, and anchorage-independent growth. The tumorigenicity of these cancer cell lines as determined by subcutaneous inoculation in nude mice was similarly suppressed by TGFBI expression. Likewise, TGFBI expression reduced the proportion of S-phase while increased the proportion of G1 phase in these cells. The redistribution of cell cycle phase after re-expression of TGFBI was correspondent with transiently elevated expression of p21 and p53. The activities of senescence-associated β-galactosidase and telomerase were enhanced in TGFBI-transfected cells.

Conclusion

Collectively, these results imply that TGFBI plays a suppressive role in the development of mesothelioma and breast cancer cells, possibly through inhibitions of cell proliferation, delaying of G1-S phase transition, and induction of senescence.

【 授权许可】

   
2012 Li et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Gibson MA, Kumaratilake JS, Cleary EG: The protein components of the 12-nanometer microfibrils of elastic and nonelastic tissues. J Biol Chem 1989, 264:4590-4598.
• [2]Skonier J, Neubauer M, Madisen L, Bennett K, Plowman GD, Purchio AF: cDNA cloning and sequence analysis of betaig-h3, a novel gene induced in a human adenocarcinoma cell line after treatment with transforming growth factor-beta. DNA Cell Biol 1992, 11:511-522.
• [3]Skonier J, Bennett K, Rothwell V, Kosowski S, Plowman G, Wallace P, Edelhoff S, Disteche C, Neubauer M, Marquardt H: betaig-h3: a transforming growth factor-beta-responsive gene encoding a secreted protein that inhibits cell attachment in vitro and suppresses the growth of CHO cells in nude mice. DNA Cell Biol 1994, 13:571-584.
• [4]Kim JE, Kim EH, Han EH, Park RW, Park IH, Jun SH, Kim JC, Young MF, Kim IS: A TGF-beta-inducible cell adhesion molecule, betaig-h3 is downregulated in melorheostosis and involved in osteogenesis. J Cell Biochem 2000, 77:169-178.
• [5]Park SW, Bae JS, Kim KS, Park SH, Lee BH, Choi JY, Park JY, Ha SW, Kim YL, Kwon TH, Kim IS, Park RW: Betaig-h3 promotes renal proximal tubular epithelial cell adhesion, migration and proliferation through the interaction with alpha3beta1 integrin. Exp Mol Med 2004, 36:211-219.
• [6]Bae JS, Lee SH, Kim JE, Choi JY, Park RW, Yong Park J, Park HS, Sohn YS, Lee DS, Bae Lee E, Kim IS: Betaig-h3 supports keratinocyte adhesion, migration, and proliferation through alpha3beta1 integrin. Biochem Biophys Res Commun 2002, 294:940-948.
• [7]Kim JE, Kim SJ, Jeong HW, Lee BH, Choi JY, Park RW, Park JY, Kim IS: RGD peptides released from betaig-h3, a TGF-beta-induced cell-adhesive molecule, mediate apoptosis. Oncogene 2003, 22:2045-2053.
• [8]Jeong HW, Kim IS: TGF-beta1 enhances betaig-h3-mediated keratinocyte cell migration through the alpha3beta1 integrin and PI3K. J Cell Biochem 2004, 92:770-780.
• [9]Kim MO, Yun SJ, Kim IS, Sohn S, Lee EH: Transforming growth factor-beta-inducible gene-h3 (beta(ig)-h3) promotes cell adhesion of human astrocytoma cells in vitro: implication of alpha6beta4 integrin. Neurosci Lett 2003, 336:93-96.
• [10]Lee BH, Bae JS, Park RW, Kim JE, Park JY, Kim IS: betaig-h3 triggers signaling pathways mediating adhesion and migration of vascular smooth muscle cells through alphavbeta5 integrin. Exp Mol Med 2006, 38:153-161.
• [11]Nam JO, Kim JE, Jeong HW, Lee SJ, Lee BH, Choi JY: V: Identification of the alphavbeta3 integrin-interacting motif of betaig-h3 and its anti-angiogenic effect. J Biol Chem 2003, 278:25902-25909.
• [12]Romero P, Vogel M, Diaz JM, Romero MP, Herrera L: Anticipation in familial lattice corneal dystrophy type I with R124C mutation in the TGFBI (BIGH3) gene. Mol Vis 2008, 14:829-835.
• [13]Poulaki V, Colby K: Genetics of anterior and stromal corneal dystrophies. Semin Ophthalmol 2008, 23:9-17.
• [14]Zhao Y, Shao G, Piao CQ, Berenguer J, Hei TK: Down-regulation of Betaig-h3 gene is involved in the tumorigenesis in human bronchial epithelial cells induced by heavy-ion radiation. Radiat Res 2004, 162:655-659.
• [15]Shao G, Berenguer J, Borczuk AC, Powell CA, Hei TK, Zhao Y: Epigenetic inactivation of Betaig-h3 gene in human cancer cells. Cancer Res 2006, 66:4566-4573.
• [16]Zhao YL, Piao CQ, Hei TK: Downregulation of Betaig-h3 gene is causally linked to tumorigenic phenotype in asbestos treated immortalized human bronchial epithelial cells. Oncogene 2002, 21:7471-7477.
• [17]Ahmed AA, Mills AD, Ibrahim AE, Temple J, Blenkiron C, Vias M, Massie CE, Iyer NG, McGeoch A, Crawford R, Nicke B, Downward J, et al.: The extracellular matrix protein TGFBI induces microtubule stabilization and sensitizes ovarian cancers to paclitaxel. Cancer Cell 2007, 12:514-527.
• [18]Robinson BW, Lake RA: Advances in malignant mesothelioma. N Engl J Med 2005, 353:1591-1603.
• [19]Peto J, Decarli A, La Vecchia C, Levi F, Negri E: The European mesothelioma epidemic. Br J Cancer 1999, 79:666-672.
• [20]Ries L, Melbert D, Krapcho M, Mariotto A, Miller BA, Feuer EJ, Clegg L, Horner MJ, Howlader N, Eisner MP, Reichman M, Edwards BK: SEER Cancer Statistics Review. National Cancer Institute, Bethesda; 2007:1975-2004.
• [21]Grushko TA, Anne Blackwood MA, Schumm PL, Hagos FG, Adeyanju MO, Feldman Michael D, Sanders MO, Weber BL, Olufunmilayo I, Olopade OI: Molecular-cytogenetic analysis of HER-2/neu Gene in BRCA1-associated breast cancers. Cancer Res 2002, 62:1481-1488.
• [22]Ellsworth RE, Deyarmin B, Hooke JA, Shriver CD: Clinical implications of the molecular relationship between HER2 and BRCA1. 2007 ASCO Annual Meeting Proceedings (Post-Meeting Edition). J Clin Oncol 2007, 25:10536.
• [23]Romero Q, Bendah PO, Klintman M, Loman N, Ingvar C, Rydén L, Rose C, Grabau D, Borgquist S: Ki67 proliferation in core biopsies versus surgical samples - a model for neo-adjuvant breast cancer studies. BMC Cancer 2011, 11:341-353. BioMed Central Full Text
• [24]Braig M, Lee S, Loddenkemper C, Rudolph C, Peters AH, Schlegelberger B, Stein H, Dörken B, Jenuwein T, Schmitt CA: Oncogene-induced senescence as an initial barrier in lymphoma development. Nature 2005, 7051:660-665.
• [25]Chen Z, Trotman LC, Shaffer D, Lin HK, Dotan ZA, Niki M, Koutcher JA, Scher HI, Ludwig T, Gerald W, Cordon-Cardo C, Pandolfi PP: Crucial role of p53-dependent cellular senescence in suppression of Pten-deficient tumorigenesis. Nature 2005, 7051:725-630.
• [26]Collado M, Gil J, Efeyan A, Guerra C, Schuhmacher AJ, Barradas M, Benguría A, Zaballos A, Flores JM, Barbacid M, Beach D, Serrano M: Tumour biology: senescence in premalignant tumours. Nature 2005, 7051:642.
• [27]Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereira-Smith O, et al.: A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Nartl Acad Sci U S A 1995, 92:9363-9367.
• [28]Muntoni A, Reddel RR: The first molecular details of ALT in human tumor cells. Hum Mol Genet 2005, 14:R191-R196.
• [29]Zhao Y, El-Gabry M, Hei TK: Loss of Betaig-h3 protein is frequent in primary lung carcinoma and related to tumorigenic phenotype in lung cancer cells. Mol Carcinog 2006, 45:84-92.
• [30]Genini M, Schwalbe P, Scholl FA, Schafer BW: Isolation of genes differentially expressed in human primary myoblasts and embryonal rhabdomyosarcoma. Int J Cancer 1996, 66:571-577.
• [31]Nabokikh A, Ilhan A, Bilban M, Gartner W, Vila G, Niederle B, Nielsen JH, Wagner O, Base W, Luger A, Wagner L: Reduced TGF-beta1 expression and its target genes in human insulinomas. Exp Clin Endocrinol Diabetes 2007, 115:674-682.
• [32]Calaf GM, Echiburu-Chau C, Zhao YL, Hei TK: BigH3 protein expression as a marker for breast cancer. Int J Mol Med 2008, 21:561-568.
• [33]Buckhaults P, Rago C, St Croix B, Romans KE, Saha S, Zhang L, Vogelstein B, Kinzler KW: Secreted and cell surface genes expressed in benign and malignant colorectal tumors. Cancer Res 2001, 61:6996-7001.
• [34]Schorderet DF, Menasche M, Morand S, Bonnel S, Buchillier V, Marchant D, Auderset K, Bonny C, Abitbol M, Munier FL: Genomic characterization and embryonic expression of the mouse Bigh3 (Tgfbi) gene. Biochem Biophys Res Commun 2000, 274:267-274.
• [35]Billings PC, Herrick DJ, Kucich U, Engelsberg BN, Abrams WR, Macarak EJ, Rosenbloom J, Howard PS: Extracellular matrix and nuclear localization of betaig-h3 in human bladder smooth muscle and fibroblast cells. J Cell Biochem 2000, 79:261-273.
• [36]Nam JO, Jeong HW, Lee BH, Park RW, Kim IS: Regulation of tumor angiogenesis by fastatin, the fourth FAS1 domain of betaig-h3, via alphavbeta3 integrin. Cancer Res 2005, 65:4153-4161.
• [37]Wen G, Hong M, Li B, Liao W, Cheng SK, Burong Hu, Calaf GM, Ping Lu, Partrodge MA, Tong J, Hei TK: Transforming growth factor-β-induced protein (TGFBI) suppresses mesothelioma progression through the Akt/mTOR pathway. Int J Oncol 2011, 39:1001-1009.
• [38]Ji P, Agrawal S, Diederichs S, Bäumer N, Becker A, Cauvet T, Kowski S, Beger C, Welte K, Berdel WE, Serve H, Müller-Tidow C: Cyclin A1, the alternative A-type cyclin, contributes to G1/S cell cycle progression in somatic cells. Oncogene 2005, 24:2739-2744.
• [39]Zhang Y, Wen G, Shao G, Wang C, Lin C, Fang H, Balajee AS, Bhagat G, Hei TK, Zhao Y: TGFBI deficiency predisposes mice to spontaneous tumor development. Cancer Res 2009, 69:37-44.
• [40]Macieira-Coelho A: Markers of 'cell senescence'. Mech Ageing Dev 1998, 104:207-211.
• [41]Collado M, Blasco MA, Serrano M: Cellular senescence in cancer and aging. Cell 2007, 130:223-233.
• [42]Hunter T, Pines J: Cyclins and cancer. II: Cyclin D and CDK inhibitors come of age. Cell 1994, 79:573-582.
• [43]Bodnar AG, Ouellete M, Frolkis M, Holt SE, Chiu C, Morin GB, Harley CB, Shay JW, Lichtsteiner S, Wright WE: Extension of life-span by introduction of telomerase into normal human cells. Science 1998, 279:349-352.
• [44]Greenberg RA, Chin L, Femino A, Lee KH, Gottlieb GJ, Singer RH, Greider CW, DePinho RA: Short dysfunctional telomeres impair tumorigenesis in the INK4aD2/3 cancer-prone mouse. Cell 1999, 97:515-525.
• [45]Halvorsen TL, Leibowitz G, Levine F: Telomerase activity is sufficient to allow transformed cells to escape from crisis. Mol Cell Biol 1999, 19:1864-1870.
• [46]Cheng JQ, Jhanwar SC, Klein WM, Bell DW, Lee WC, Altomare DA, et al.: p16 alterations and deletion mapping of 9p21-p22 in malignant mesothelioma. Cancer Res 1994, 54:5547-5551.
• [47]Usami N, Sekido Y, Maeda O, Yamamoto K, Minna JD, Hasegawa Y, Nobori T, Olopade OI, Buckler AJ, Testa JR: Beta-catenin inhibits cell growth of a malignant mesothelioma cell line, NCI-H28, with a 3p21.3 homozygous deletion. Oncogene 2003, 22:7923-7930.
• [48]Petroulakis E, Parsyan A, Dowling RJ, LeBacquer O, Martineau Y, Bidinosti M, Larsson O, Alain T, Rong L, Mamane Y, Paquet M, Furic L, Topisirovic I, Shahbazian D, Livingstone M, Costa-Mattioli M, Teodoro JG, Sonenberg N: p53-dependent translational control of senescence and transformation via 4E-BPs. Cancer Cell 2009, 16:439-446.
• [49]Noda A, Ning Y, Venable SF, Pereira-Smith OM, Smith JR: Cloning of senescent cell-derived inhibitors of DNA synthesis using an expression screen. Exp Cell Res 1994, 211:90-98.
• [50]Urruticoechea A, Smith IE, Dowsett M: Proliferation marker Ki-67 in early breast cancer. J Clin Oncol 2005, 23:7212-7220.
• [51]Ma C, Rong Y, Radiloff DR, Datto MB, Centeno B, Bao S, Cheng AW, Lin F, Jiang S, Yeatman TJ, Wang XF: Extracellular matrix protein betaig-h3/TGFBI promotes metastasis of colon cancer by enhancing cell extravasation. Genes Dev 2008, 22:308-321.
• [52]Ween MP, Lokman NA, Hoffmann P, Rodgers RJ, Ricciardelli C, Oehler MK: Transforming growth factor-beta-induced protein secreted by peritoneal cells increases the metastatic potential of ovarian cancer cells. Intl J Cancer 2011, 128:1570-1584.
• [53]Schneider D, Kleeff J, Berberat PO, Zhu Z, Korc M, Friess H, Buchler MW: Induction and expression of BigH3 in pancreatic cancer cells. Biochem. Biophys. ACTA 2007, 1588:1-6.
• [54]Yamanaka M, Kimura F, Kagata Y, Kondoh N, Asano T, Yamamoto M, Hayakawa M: BigH3 is overexpressed in clear cell renal cell carcinoma. Oncology Report 2008, 19:865-874.
• [55]Lin B, Madan A, Yoon JG, Fang X, Yan X, Kim TK, Hwang D, Hood L, Foltz G: Massively parallel signature sequencing and bioinformatics analysis identifies up-regulation of TGFBI and SOX4 in human glioblastoma. PLoS One 2010, 4:e10210.