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Molecular Cancer

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Open Access 01-12-2010 | Research

Epigenetic silencing of MAL, a putative tumor suppressor gene, can contribute to human epithelium cell carcinoma

Authors: Wei Cao, Zhi-yuan Zhang, Qin Xu, Qiang Sun, Ming Yan, Jun Zhang, Ping Zhang, Ze-guang Han, Wan-tao Chen

Published in: Molecular Cancer | Issue 1/2010

Abstract

Background

To identify new and useful candidate biomarkers in head and neck squamous cell carcinoma (HNSCC), we performed a genome-wide survey and found that Myelin and lymphocyte-associated protein (MAL) was a gene that was markedly down-regulated in HNSCC. Hence, we investigated the mechanism of MAL silencing and the effects of MAL on the proliferation, invasion, and apoptotic potential in HNSCC.

Results

MAL was significantly down-regulated in 91.7% of HNSCC specimens at the mRNA level as compared with adjacent normal tissues (P = 0.0004). Moreover, the relative transcript levels of the MAL gene were remarkably decreased by five-fold in nine HNSCC cell lines as compared with normal head and neck epithelium cells. MAL gene expression was restored in 44%, 67%, and 89% in HNSCC cell lines treated with TSA, 5-Aza-dC, and TSA plus 5-Aza-dC, respectively. Furthermore, bisulfate-treated DNA sequencing demonstrated that the two CpG islands (that is, M₁ and M₂) located in MAL promoter region were completely methylated in the HNSCC cell lines (CpG methylated ratio was more than 90%), and only one CpG island (that is, M₁) was partially methylated in HNSCC tissues (CpG methylated ratio between 20% and 90%). A significant reduction in cell proliferation and a change in the cell cycle profile were also observed in MAL transfectants. Matrigel assay demonstrated that the invasiveness of HNSCC cells significantly decreased. A significant increase in the population of apoptotic cells was observed in MAL transfected cells. The exogenous expression of the MAL gene suppressed malignant phenotypes, while the cell death induced by MAL gene transfer was a result of apoptosis as demonstrated by the induction of cleavage of the poly (that is, ADP-ribose) polymerase. Additionally, tumor growth was suppressed in cells expressing MAL as compared with cells not expressing MAL.

Conclusion

Our data suggest that the epigenetic inactivation of MAL, as a candidate tumor suppressor gene, can contribute to human epithelial cell carcinoma and may be served as a biomarker in HNSCC.

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Mignogna MD, Fedele S, Lo Russo L: The World Cancer Report and the burden of oral cancer. Eur J Cancer Prev. 2004, 13 (2): 139-142. 10.1097/00008469-200404000-00008CrossRefPubMed

Mohr S, Leikauf GD, Keith G, Rihn BH: Microarrays as cancer keys: an array of possibilities. J Clin Oncol. 2002, 20 (14): 3165-3175. 10.1200/JCO.2002.12.073CrossRefPubMed

Alevizos I, Mahadevappa M, Zhang X, Ohyama H, Kohno Y, Posner M, Gallagher GT, Varvares M, Cohen D, Kim Dl: Oral cancer in vivo gene expression profiling assisted by laser capture microdissection and microarray analysis. Oncogene. 2001, 20 (43): 6196-6204. 10.1038/sj.onc.1204685CrossRefPubMed

Belbin TJ, Singh B, Smith RV, Socci ND, Wreesmann VB, Sanchez-Carbayo M, Masterson J, Patel S, Cordon-Cardo C, Prystowsky MB: Molecular profiling of tumor progression in head and neck cancer. Archives of otolaryngology-head & neck surgery. 2005, 131 (1): 10-18.CrossRef

Carles A, Millon R, Cromer A, Ganguli G, Lemaire F, Young J, Wasylyk C, Muller D, Schultz I, Rabouel Y: Head and neck squamous cell carcinoma transcriptome analysis by comprehensive validated differential display. Oncogene. 2006, 25 (12): 1821-1831. 10.1038/sj.onc.1209203CrossRefPubMed

Chin D, Boyle GM, Williams RM, Ferguson K, Pandeya N, Pedley J, Campbell CM, Theile DR, Parsons PG, Coman WB: Novel markers for poor prognosis in head and neck cancer. Int J Cancer. 2005, 113 (5): 789-797. 10.1002/ijc.20608CrossRefPubMed

Kuriakose MA, Chen WT, He ZM, Sikora AG, Zhang P, Zhang ZY, Qiu WL, Hsu DF, McMunn-Coffran C, Brown SM, Elango EM, Delacure MD, Chen FA: Selection and validation of differentially expressed genes in head and neck cancer. CMLS, Cell Mol Life Sc. 2004, 61 (11): 1372-1383. 10.1007/s00018-004-4069-0.CrossRef

Mimori K, Shiraishi T, Mashino K, Sonoda H, Yamashita K, Yoshinaga K, Masuda T, Utsunomiya T, Alonso MA, Inoue H, Mori M: MAL gene expression in esophageal cancer suppresses motility, invasion and tumorigenicity and enhances apoptosis through the Fas pathway. Oncogene. 2003, 22 (22): 3463-3471. 10.1038/sj.onc.1206378CrossRefPubMed

Lind GE, Ahlquist T, Kolberg M, Berg M, Eknaes M, Alonso MA, Kallioniemi A, Meling GI, Skotheim RI, Rognum TO, Thiis-Evensen E, Lothe RA: Hypermethylated MAL gene - a silent marker of early colon tumorigenesis. J Transl Med. 2008, 6: 13- 10.1186/1479-5876-6-13PubMedCentralCrossRefPubMed

10.

Buffart TE, Overmeer RM, Steenbergen RD, Tijssen M, Van Grieken NC, Snijders PJ, Grabsch HI, Van de Velde CJ, Carvalho B, Meijer GA: MAL promoter hypermethylation as a novel prognostic marker in gastric cancer. Br J Cancer. 2008, 99 (11): 1802-1807. 10.1038/sj.bjc.6604777PubMedCentralCrossRefPubMed

11.

Horne HN, Lee PS, Murphy SK, Alonso MA, Olson JA, Marks JR: Inactivation of the MAL gene in breast cancer is a common event that predicts benefit from adjuvant chemotherapy. Mol Cancer Res. 2009, 7 (2): 199-209. 10.1158/1541-7786.MCR-08-0314PubMedCentralCrossRefPubMed

12.

Hatta M, Nagai H, Okino K, Onda M, Yoneyama K, Ohta Y, Nakayama H, Araki T, Emi M: Down-regulation of members of glycolipid-enriched membrane raft gene family, MAL and BENE, in cervical squamous cell cancers. J Obstet Gynaecol Res. 2004, 30 (1): 53-58. 10.1111/j.1341-8076.2004.00156.xCrossRefPubMed

13.

Maruya S, Kim HW, Weber RS, Lee JJ, Kies M, Luna MA, Batsakis JG, El-Naggar AK: Gene expression screening of salivary gland neoplasms: molecular markers of potential histogenetic and clinical significance. J Mol Diagn. 2004, 6 (3): 180-90.PubMedCentralCrossRefPubMed

14.

Alonso MA, Weissman SM: cDNA cloning and sequence of MAL, a hydrophobic protein associated with human T-cell differentiation. Proc Natl Acad Sic USA. 1987, 84 (7): 1997-2001. 10.1073/pnas.84.7.1997.CrossRef

15.

Kim T, Fiedler K, Madison DL, Krueger WH, Pfeiffer SE: Cloning and characterization of MVP17: a developmentally regulated myelin protein in oligodendrocytes. J Neurosci Res. 1995, 42 (3): 413-422. 10.1002/jnr.490420316CrossRefPubMed

16.

Simons K, Ikonen E: Functional rafts in cell membranes. Nature. 1997, 387 (6633): 569-572. 10.1038/42408CrossRefPubMed

17.

Cheong KH, Zacchetti D, Schneeberger EE, Simons K: VIP17/MAL, a lipid raft-associated protein, is involved in apical transport in MDCK cells. Pro Natl Acad Sci USA. 1999, 96 (11): 6241-6248. 10.1073/pnas.96.11.6241.CrossRef

18.

Puertollano R, Martín-Belmonte F, Millán J, de Marco MC, Albar JP, Kremer L, Alonso MA: The MAL proteolipid is necessary for normal apical transport and accurate sorting of the influenza virus hemagglutinin in Madin-Darby canine kidney cells. J Cell Biol. 1999, 145 (1): 141-151. 10.1083/jcb.145.1.141PubMedCentralCrossRefPubMed

19.

Wong YF, Cheung TH, Tsao GS, Lo KW, Yim SF, Wang VW, Heung MM, Chan SC, Chan LK, Ho TW, Wong KW, Guo Y, Chung TK, Smith DI: Genome-wide gene expression profiling of cervical cancer in Hong Kong women by oligonucleotide microarray. Int J Cancer. 2006, 118 (10): 2461-2469. 10.1002/ijc.21660CrossRefPubMed

20.

Mori Y, Cai K, Cheng Y, Wang S, Paun B, Hamilton JP, Jin Z, Sato F, Berki AT, Kan T, Ito T, Mantzur C, Abraham JM, Meltzer SJ: A Genome-wide search identifies epigenetic sliencing of somatostain, tachykinin-1 and 5 other genes in colon cancer. Gastroenterology. 2006, 131 (3): 797-808. 10.1053/j.gastro.2006.06.006CrossRefPubMed

21.

Beder LB, Gunduz M, Hotomi M, Fujihara K, Shimada J, Tamura S, Gunduz E, Fukushima K, Yaykasli K, Grenman R, Shimizu K, Yamanaka N: T-lymphocyte maturation-associated protein gene as a candidate metastasis suppressor for head and neck squamous cell carcinomas. Cancer Sci. 2009, 100 (5): 873-880. 10.1111/j.1349-7006.2009.01132.xCrossRefPubMed

22.

Berchuck A, Iversen ES, Luo J, Clarke JP, Horne H, Levine DA, Boyd J, Alonso MA, Secord AA, Bernardini MQ, Barnett JC, Boren T, Murphy SK, Dressman HK, Marks JR, Lancaster JM: Microarray analysis of early stage serous ovarian cancers shows profiles predictive of favorable outcome. Clin Cancer Res. 2009, 15 (7): 2448-2455. 10.1158/1078-0432.CCR-08-2430PubMedCentralCrossRefPubMed

23.

Rancaño C, Rubio T, Alonso MA: Alternative splicing of human T-cell-specific MAL mRNA and its correlation with the exon/intron organization of the gene. Genomics. 1994, 21 (2): 447-450. 10.1006/geno.1994.1294CrossRefPubMed

24.

Herman JG, Graff JR, Myöhänen S, Nelkin BD, Baylin SB: Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc Natl Acad Sci USA. 1996, 93 (18): 9821-9826. 10.1073/pnas.93.18.9821PubMedCentralCrossRefPubMed

25.

Boulares AH, Yakovlev AG, Ivanova V, Stoica BA, Wang G, Iyer S, Smulson M: Role of poly(ADP-ribose) polymerase (PARP) cleavage in apoptosis. Caspase 3-resistant PARP mutant increases rates of apoptosis in transfected cells. J Biol Chem. 1999, 274 (33): 22932-22940. 10.1074/jbc.274.33.22932CrossRefPubMed

26.

Tomioka H, Morita K, Hasegawa S, Omura K: Gene expression analysis by cDNA microarray in oral squamous cell carcinoma. J Oral Pathol Med. 2006, 35 (4): 206-211. 10.1111/j.1600-0714.2006.00410.xCrossRefPubMed

27.

Ziober AF, Patel KR, Alawi F, Gimotty P, Weber RS, Feldman MM, Chalian AA, Weinstein GS, Hunt J, Ziober BL: Identification of a gene signature for rapid screening of oral squamous cell carcinoma. Clin Cancer Res. 2006, 12 (20 Pt 1): 5960-5971. 10.1158/1078-0432.CCR-06-0535CrossRefPubMed

28.

Ye H, Yu T, Temam S, Ziober BL, Wang J, Schwartz JL, Mao L, Wong DT, Zhou X: Transcriptomic dissection of tongue squamous cell carcinoma. BMC Genomics. 2008, 9: 69- 10.1186/1471-2164-9-69PubMedCentralCrossRefPubMed

29.

Choi P, Jordan CD, Mendez E, Houck J, Yueh B, Farwell DG, Futran N, Chen C: Examination of oral cancer biomarkers by tissue microarray analysis. Arch Otolaryngol Head Neck Surg. 2008, 134 (5): 539-546. 10.1001/archotol.134.5.539PubMedCentralCrossRefPubMed

30.

Marte B, Eccleston A, Nath D: Molecular cancer diagnostics. Nature. 2008, 452 (7187): 547- 10.1038/452547aCrossRefPubMed

31.

Lallemant B, Evrard A, Combescure C, Chapuis H, Chambon G, Raynal C, Reynaud C, Sabra O, Joubert D, Hollande F, Lallemant JG, Lumbroso S, Brouillet JP: Clinical relevance of nine transcriptional molecular markers for the diagnosis of head and neck squamous cell carcinoma in tissue and saliva rinse. BMC Cancer. 2009, 9: 370- 10.1186/1471-2407-9-370PubMedCentralCrossRefPubMed

32.

Martín-Belmonte F, Arvan P, Alonso MA: MAL mediates apical transport of secretory proteins in polarized epithelial Madin-Darby canine kidney cells. J Biol Chem. 2001, 276 (52): 49337-49342. 10.1074/jbc.M106882200CrossRefPubMed

33.

Martín-Belmonte F, Puertollano R, Millán J, Alonso MA: The MAL proteolipid is necessary for the overall apical delivery of membrane proteins in the polarized epithelial Madin-Darby canine kidney and fischer rat thyroid cell lines. Mol Biol Cell. 2000, 11 (6): 2033-2045.PubMedCentralCrossRefPubMed

34.

O'Regan EM, Toner ME, Finn SP, Fan CY, Ring M, Hagmar B, Timon C, Smyth P, Cahill S, Flavin R, Sheils OM, O'Leary JJ: p16(INK4A) genetic and epigenetic profiles differ in relation to age and site in head and neck squamous cell carcinomas. Hum Pathol. 2008, 39 (3): 452-458. 10.1016/j.humpath.2007.08.004CrossRefPubMed

35.

Konrad M, Saunier S, Heidet L, Silbermann F, Benessy F, Calado J, Le Paslier D, Broyer M, Gubler MC, Antignac C: Large homozygous deletions of the 2q13 region are a major cause of juvenile nephronophthisis. Hum Mol Genet. 1996, 5 (3): 367-371. 10.1093/hmg/5.3.367CrossRefPubMed

36.

Overmeer RM, Henken FE, Bierkens M, Wilting SM, Timmerman I, Meijer CJ, Snijder PJ, Steenbergen RD: Repression of MAL tumour suppressor activity by promoter methylation during cervical carcinogenesis. J Pathol. 2009, 219 (3): 327-336. 10.1002/path.2598CrossRefPubMed

37.

Lee PS, Teaberry VS, Bland AE, Huang Z, Whitaker RS, Baba T, Fujii S, Secord AA, Berchuck A, Murphy SK: Elevated MAL expression is accompanied by promoter hypomethylation and platinum resistance in epithelial ovarian cancer. Int J Cancer. 2010, 126 (6): 1378-1389.PubMed

38.

Ahlquist T, Lind GE, Costa VL, Meling GI, Vatn M, Hoff GS, Rognum TO, Skotheim RI, Thiis-Evensen E, Lothe RA: Gene methylation profiles of normal mucosa, and benign and malignant colorectal tumors identify early onset markers. Mol Cancer. 2008, 7: 94- 10.1186/1476-4598-7-94PubMedCentralCrossRefPubMed

39.

Sobin LH, Fleming ID: TNM Classification of Malignant Tumors, fifth edition (1997). Union Internationale Contre le Cancer and the American Joint Committee on Cancer. Cancer. 1997, 80 (9): 1803-1804. 10.1002/(SICI)1097-0142(19971101)80:9<1803::AID-CNCR16>3.0.CO;2-9CrossRefPubMed

40.

Loiaconi G, Berth AF: Current aspects of histological typing of carcinomas of the oral cavity. Riv Ital Stomatol. 1982, 51 (5): 415-421.PubMed

Title: Epigenetic silencing of MAL, a putative tumor suppressor gene, can contribute to human epithelium cell carcinoma
Authors: Wei Cao
Zhi-yuan Zhang
Qin Xu
Qiang Sun
Ming Yan
Jun Zhang
Ping Zhang
Ze-guang Han
Wan-tao Chen
Publication date: 01-12-2010
Publisher: BioMed Central
Published in: Molecular Cancer / Issue 1/2010
Electronic ISSN: 1476-4598
DOI: https://doi.org/10.1186/1476-4598-9-296

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