Abstract
Array-based comparative genomic hybridization (array-CGH) has good potential for the high-throughput identification of genetic aberrations in cell genomes. In the course of a program to screen a panel of oral squamous-cell carcinoma (OSCC), cell lines for genomic copy-number aberrations by array-CGH using our in-house arrays, we identified a 3-Mb homozygous deletion at 10p12 in 1 of 18 cell lines (5.6%). Among seven genes located within this region, expression of PRTFDC1 mRNA was not detected in 50% (9/18) or decreased in 5.6% (1/18) of OSCC cell lines, but detected in normal oral epithelia and restored in gene-silenced OSCC cells without its homozygous loss after treatment with 5-aza-2′-deoxycytidine. Among 17 cell lines without a homozygous deletion, the hypermethylation of the PRTFDC1 CpG island, which showed promoter activity, was observed in all nine cell lines with no or reduced PRTFDC1 expression (52.9%). Methylation of this CpG island was also observed in primary OSCC tissues (8/47, 17.0%). In addition, restoration of PRTFDC1 in OSCC cells lacking its expression inhibited cell growth in colony-formation assays, whereas knockdown of PRTFDC1 expression in OSCC cells expressing the gene promoted cell growth. These results suggest that epigenetic silencing of PRTFDC1 by hypermethylation of the CpG island leads to a loss of PRTFDC1 function, which might be involved in squamous cell oral carcinogenesis.
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Akanuma D, Uzawa N, Yoshida MA, Negishi A, Amagasa T, Ikeuchi T . (1999). Inactivation patterns of the p16 (INK4a) gene in oral squamous cell carcinoma cell lines. Oral Oncol 35: 476–483.
Baldwin C, Garnis C, Zhang L, Rosin MP, Lam WL . (2005). Multiple microalterations detected at high frequency in oral cancer. Cancer Res 65: 7561–7567.
Brinkman BM, Wong DT . (2006). Disease mechanism and biomarkers of oral squamous cell carcinoma. Curr Opin Oncol 18: 228–233.
Friend SH, Bernards R, Rogelj S, Weinberg RA, Rapaport JM, Albert DM et al. (1986). A human DNA segment with properties of the gene that predisposes to retinoblastoma and osteosarcoma. Nature (London) 323: 643–646.
Fujimoto R, Kamata N, Yokoyama K, Taki M, Tomonari M, Tsutsumi S et al. (2002). Establishment of immortalized human oral keratinocytes by gene transfer of a telomerase component. J Jpn Oral Muco Membr (Japanese) 8: 1–8.
Ha PK, Califano JA . (2006). Promoter methylation and inactivation of tumour-suppressor genes in oral squamous-cell carcinoma. Lancet Oncol 7: 77–82.
Hahn SA, Schutte M, Hoque AT, Moskaluk CA, da Costa LT, Rozenblum E et al. (1996). DPC4, a candidate tumor suppressor gene at human chromosome 18q21.1. Science (Wash. DC) 271: 350–353.
Hermsen MA, Joenje H, Arwert F, Braakhuis BJ, Baak JP, Westerveld A et al. (1997). Assessment of chromosomal gains and losses in oral squamous cell carcinoma by comparative genomic hybridisation. Oral Oncol 33: 414–418.
Imoto I, Izumi H, Yokoi S, Hosoda H, Shibata T, Hosoda F et al. (2006). Frequent silencing of the candidate tumor suppressor PCDH20 by epigenetic mechanism in non-small-cell lung cancers. Cancer Res 66: 4617–4626.
Inazawa J, Inoue J, Imoto I . (2004). Comparative genomic hybridization (CGH)-arrays pave the way for identification of novel cancer-related genes. Cancer Sci 95: 559–563.
Izumi H, Inoue J, Yokoi S, Hosoda H, Shibata T, Sunamori M et al. (2005). Frequent silencing of DBC1 is by genetic or epigenetic mechanisms in non-small cell lung cancers. Hum Mol Genet 14: 997–1007.
Jin C, Jin Y, Wennerberg J, Annertz K, Enoksson J, Mertens F . (2006). Cytogenetic abnormalities in 106 oral squamous cell carcinomas. Cancer Genet Cytogenet 164: 44–53.
Jones PA, Baylin SB . (2002). The fundamental role of epigenetics events in cancer. Nat Rev Genet 3: 415–428.
Kamb A, Gruis NA, Weaver-Feldhaus J, Liu Q, Harshman K, Tavtigian SV et al. (1994). A cell cycle regulator potentially involved in genesis of many tumor types. Science (Wash DC) 264: 436–440.
Kolb A . (2003). The first intron of the murine β-casein gene contains a functional promoter. Biochem Biophys Res Commun 306: 1099–1105.
Kusumoto Y, Hirano H, Saitoh K, Yamada S, Takedachi M, Nozaki T et al. (2004). Human gingival epithelial cells produce chemotactic factors Interleukin-8 and Monocyte Chemoattractant Protein-1 after stimulation with Porphyromonas gingivalis via Toll-Like Receptor 2. J Perioodontl 75: 370–378.
Li J, Yen C, Liaw D, Podsypanina K, Bose S, Wang SI et al. (1997). PTEN, a putative protein tyrosine phosphatase gene mutated in human brain, breast, and prostate cancer. Science (Wash. DC) 275: 1943–1947.
Misawa A, Inoue J, Sugino Y, Hosoi H, Sugimoto T, Hosoda F et al. (2005). Methylation-associated silencing of the nuclear receptor 1I2 gene in advanced-type neuroblastomas, identified by bacterial artificial chromosome array-based methylated CpG island amplicon. Cancer Res 65: 10233–10242.
Nagpal JK, Das BR . (2003). Oral cancer: reviewing the present understanding of its molecular mechanism and exploring the future directions for its effective management. Oral Oncol 39: 213–221.
Nakagawachi T, Soejima H, Urano T, Zhao W, Higashimoto K, Satoh Y et al. (2003). Silencing effect of CpG island hypermethylation and histone modifications on O6-methylguanine-DNA methyltransferase (MGMT) gene expression in human cancer. Oncogene 22: 8835–8844.
Nicklas JA . (2006). Pseudogenes of the human HPRT1 gene. Environ Mol Mutagen 47: 212–218.
Parkin DM, Bray F, Ferlay J, Pisani P . (2005). Global cancer statistics, 2002. CA Cancer J Clin 55: 74–108.
Saigusa K, Imoto I, Tanikawa C, Aoyagi M, Ohno K, Nakamura Y et al. (2007). RGC32, a novel p53-inducible gene, is located on centrosomes during mitosis and results in G2/M arrest. Oncogene 26: 1110–1121.
Scully C, Field JK, Tanzawa H . (2000). Genetic aberrations in oral or head and neck squamous cell carcinoma (SCCHN). I. Carcinogen metabolism, DNA repair and cell cycle control. Oral Oncol 36: 256–263.
Shaw R . (2006). The epigenetics of oral cancer. Int J Oral Maxillofac Surg 35: 101–108.
Snijders AM, Nowak N, Segraves R, Blackwood S, Brown N, Conroy J et al. (2001). Assembly of microarrays for genome-wide measurement of DNA copy number. Nat Genet 29: 263–264.
Snijders AM, Schmidt BL, Fridlyand J, Dekker N, Pinkel D, Jordan RC et al. (2005). Rare amplicons implicate frequent deregulation of cell fate specification pathways in oral squamous cell carcinoma. Oncogene 24: 4232–4242.
Sonoda I, Imoto I, Inoue J, Shibata T, Shimada Y, Chin K et al. (2004). Frequent silencing of low density lipoprotein receptor-related protein 1B (LRP1B) expression by genetic and epigenetic mechanisms in esophageal squamous cell carcinoma. Cancer Res 64: 3741–3747.
Takada H, Imoto I, Tsuda H, Nakanishi Y, Ichikura T, Mochizuki H et al. (2005b). ADAM23, a possible tumor suppressor gene, is frequently silenced in gastric cancers by homozygous deletion or aberrant promoter hypermethylation. Oncogene 24: 8051–8060.
Takada H, Imoto I, Tsuda H, Nakanishi Y, Sakakura C, Mitsufuji S et al. (2006). Genomic loss and epigenetic silencing of very-low-density lipoprotein receptor involved in gastric carcinogenesis. Oncogene 25: 6554–6562.
Takada H, Imoto I, Tsuda H, Sonoda I, Ichikura T, Mochizuki H et al. (2005a). Screening of DNA copy-number aberrations in gastric cancer cell lines by array-based comparative genomic hybridization. Cancer Sci 96: 100–110.
Wolff E, Girod S, Liehr T, Vorderwulbecke U, Ries J, Steininger H et al. (1998). Oral squamous cell carcinomas are characterized by a rather uniform pattern of genomic imbalances detected by comparative genomic hybridisation. Oral Oncol 34: 186–190.
Xiong Z, Laird PW . (1997). COBRA: a sensitive and quantitative DNA methylation assay. Nucleic Acids Res 25: 2532–2534.
Acknowledgements
We are grateful to Professor Yusuke Nakamura (Human Genome Center, The Institute of Medical Science, The University of Tokyo) for his continuous encouragement throughout this work. We thank Ai Watanabe, Ayako Takahashi and Rumi Mori for technical assistance. This work was supported by Grants-in-Aid for Scientific Research on Priority Areas (C) and 21st Century Center of Excellence Program for Molecular Destruction and Reconstitution of Tooth and Bone from the Ministry of Education, Culture, Sports, Science and Technology, Japan; and by a Grant-in-Aid from Core Research for Evolutional Science and Technology (CREST) of the Japan Science and Technology Corporation (JST).
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).
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Suzuki, E., Imoto, I., Pimkhaokham, A. et al. PRTFDC1, a possible tumor-suppressor gene, is frequently silenced in oral squamous-cell carcinomas by aberrant promoter hypermethylation. Oncogene 26, 7921–7932 (2007). https://doi.org/10.1038/sj.onc.1210589
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DOI: https://doi.org/10.1038/sj.onc.1210589
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