Abstract
Gene expression changes in rat asbestos-induced malignant mesothelioma (MM) cells were investigated by differential mRNA display. A mRNA transcript identified by this approach was abundant in normal rat mesothelial cells but not expressed in rat MM cell lines. Northern blot analysis confirmed that this transcript is uniformly silenced in rat MM cell lines and primary tumors. Nucleotide sequence analysis revealed that this transcript is encoded by the rat glypican 3 gene (GPC3), whose human homolog is mutated in the Simpson-Golabi-Behmel overgrowth syndrome. Allelic loss at the GPC3 locus was infrequent (6.9%) in MM cell lines, and no mutations were found. GPC3 transcript levels were markedly decreased in 16 of 18 primary tumors and 17 of 22 human MM cell lines. Most of the cell lines were shown to have aberrant methylation of the GPC3 promoter region. In two of four human MM cell lines tested, GPC3 expression was restored after 2-deoxy 5-azacytidine (DAC)-mediated demethylation of its promoter region. Ectopic expression of GPC3 inhibited in vitro colony formation of human MM cells. Collectively, these data suggest that down-regulation of GPC3 is a common occurrence in MM and that GPC3, an X-linked recessive overgrowth gene, may encode a negative regulator of mesothelial cell growth.
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Baker SJ, Markowitz S, Fearon ER, Willson JK and Vogelstein B. . 1990 Science 249: 912–915.
Bianchi AB, Mitsunaga SI, Cheng JQ, Klein WM, Jhanwar SC, Seizinger B, Kley N, Klein-Szanto AJP and Testa JR. . 1995 Proc. Natl. Acad. Sci. USA 92: 10854–10858.
Bonneh-Barkay D, Shlissel M, Berman B, Shaoul E, Admon A, Vlodavsky I, Carey DJ, Asundi VK, Reich-Slotky R and Ron D. . 1997 J. Biol. Chem. 272: 12415–12421.
Cameron EE, Bachman KE, Myohanen S, Herman JG and Baylin SB. . 1999 Nature Genet. 21: 103–107.
Cheng JQ, Jhanwar SC, Klein WM, Bell DW, Lee W-C, Altomare DA, Nobori T, Olopade OI, Buckler AJ and Testa JR. . 1994 Cancer Res. 54: 5547–5551.
Craighead JE. . 1987 Hum. Pathol. 18: 544–557.
Craighead JE, Akley NJ, Gould LB and Libbus BL. . 1987 Am. J. Pathol. 129: 448–462.
David G. . 1993 FASEB J. 7: 1023–1030.
Davis JMG. . 1974 J. Natl. Cancer Inst. 52: 1823–1829.
Filmus J, Church JG and Buick RN. . 1988 Mol. Cell Biol. 8: 4243–4249.
Filmus J, Shi W, Wong ZM and Wong M. . 1995 Biochem. J. 331: 561–565.
Funaki K, Everitt J, Bermudez E and Walker C. . 1991 Cancer Res. 51: 4059–4066.
Gonzalez AD, Kaya M, Shi W, Song H, Testa JR, Penn LZ and Filmus J. . 1998 J. Cell Biol. 141: 1407–1414.
Herman JG, Latif F, Weng Y, Lerman MI, Zbar B, Liu S, Samid D, Duan D-SR, Gnarra JR, Linehan WM and Baylin SB. . 1994 Proc. Natl. Acad. Sci. USA 91: 9700–9704.
Hill RJ, Edwards RE and Carthew P. . 1990 J. Exp. Pathol. 71: 105–118.
Huber R, Hansen RS, Strazzullo M, Pengue G, Mazzarella R, D'Urso M, Schlessinger D, Pilia G, Gartler SM and D'Esposito M. . 1999 Proc. Natl. Acad. Sci. USA 96: 616–621.
Janssen YM, Heintz NH and Mossman BT. . 1995 Cancer Res. 55: 2085–2089.
Liang P and Pardee AB. . 1992 Science 257: 967–971.
Libbus BL and Craighead JE. . 1988 Cancer Res. 48: 6455–6461.
Lin H, Huber R, Schlessinger D and Morin PJ. . 1999 Cancer Res. 59: 807–810.
Mossman BT and Gee JBL. . 1989 N. Engl. J. Med. 320: 1721–1730.
Pilia G, Hughes-Benzie RM, MacKenzie A, Baybayan P, Chen EY, Huber R, Neri G, Cao A, Forabosco A and Schlessinger D. . 1996 Nature Genet. 12: 241–247.
Sekido Y, Pass HI, Bader S, Mew DJ, Christman MF, Gazdar AF and Minna JD. . 1995 Cancer Res. 55: 1227–1231.
Song HH, Shi W and Filmus J. . 1997 J. Biol. Chem. 272: 7572–7574.
Steinfeld R, VanDen Berghe H and David G. . 1996 J. Cell Biol. 133: 405–416.
Taguchi T, Jhanwar SC, Siegfried JM, Keller SM and Testa JR. . 1993 Cancer Res. 53: 4349–4355.
Wagner JC, Berry G and Timbrell V. . 1973 Br. J. Cancer 28: 173–185.
Wagner JC, Sleggs CA and Marchland P. . 1960 Br. J. Ind. Med. 17: 260–271.
Zou Z, Anisowicz A, Hendrix MJC, Thor A, Neveu M, Sheng S, Rafidi K, Seftor E and Sager R. . 1994 Science 263: 526–529.
Acknowledgements
This report is dedicated to the memory of Ronald N Buick, Ph.D. (1948–1996) who provided reagents and valuable insights. We thank Drs Alfred Knudson, Alfonso Bellacosa and Christos Patriotis for helpful comments. Rat MM cell lines T08 and T10 were kindly provided by Dr David Coffin (USEPA, Research Triangle Park, NC, USA), and normal human mesothelial cells were a gift from Dr Edward Gabrielson (Johns Hopkins University, Baltimore, MD, USA). This study was supported by National Cancer Institute Grants CA-45745 and CA-06927, by a gift from the Local #14 Mesothelioma Fund of the International Association of Heat and Frost Insulators & Asbestos Workers in memory of Hank Vaughan and Alice Haas, and by an appropriation from the Commonwealth of Pennsylvania.
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Murthy, S., Shen, T., De Rienzo, A. et al. Expression of GPC3, an X-linked recessive overgrowth gene, is silenced in malignant mesothelioma. Oncogene 19, 410–416 (2000). https://doi.org/10.1038/sj.onc.1203322
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DOI: https://doi.org/10.1038/sj.onc.1203322
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