Promoter methylation and expression of TIMP3 gene in gastric cancer

Diagnostic Pathology

DongQiu Dai¹  ShiHui Song²  Jun Zhang²  ZhiYu Guan² 

[1] Department of Oncology, the First Affiliated Hospital of China Medical University, 155 Nanjing Beijie, Heping Section, Shenyang, Liaoning Province 110001, China;Department of Cardiovascular and Thoracic Surgery, TianJin Medical University General Hospital, 154 AnShan Road, HePing Section, TianJin 300052, China

关键词: Methylation;    Tissue Inhibitor of Metalloproteinase 3 (TIMP3);    Gastric Adenocarcinoma;   

Others  :  806308
DOI  :  10.1186/1746-1596-8-110

 received in 2013-03-28, accepted in 2013-06-29,  发布年份 2013

【 摘 要 】

Background

Gastric carcinoma development is a multi-stage process that involves more than one gene. Aberrant changes in DNA methylation are considered as the third mechanism that leads to anti-oncogene inactivation, which plays an essential role in tumor development. In this study, we assessed the relationship among the aberrant methylation of the promoter CpG islands of tissue inhibitor of metalloproteinase 3 (TIMP3) gene, its protein expression, and the clinicopathological features of gastric adenocarcinoma.

Methods

The methylation status of the promoter CpG islands and the protein expression of TIMP3 gene in tumors and adjacent normal mucosal tissues of 78 patients with gastric adenocarcinoma were detected by methylation-specific PCR (MSP) and immunohistochemistry.

Results

The CpG island methylation of TIMP3 was detected in tumor tissues, cancer-adjacent tissues, and lymph nodes with metastasis. In increasing order, the hypermethylation frequency of these tissues were 35.9% (28 of 78 non-neoplastic tissues), 85% (17 of 20 early-stage cases), 89.7% (52 of 58 progressive-stage cases), and 100% (78 of 78 metastatic lymph node). A marked difference was found between tumors and non-neoplastic tissues (P < 0.05), but no difference existed among the subgroups of tumors (P > 0.05). Immunohistochemistry analysis confirmed TIMP3 down-regulation in tumor tissues. The rate of TIMP3 gene expression was 100% in non-neoplastic tissues but apparently decreased to various extents at different stages, i.e., decreased to 30% (6/20) at the early stage, to 3.4% (2/58) at the progressive stage, and to 0% (0/78) in metastatic lymph nodes. Among the 70 tumor tissues with negative TIMP3 expression, 64 (91.4%) were hypermethylated and 6 were unmethylated (8.6%), indicating a significant association between hypermethylation and reduced or negative TIMP3 expression (P < 0.01).

Conclusion

The hypermethylation of the promoter region in CpG islands is the main mechanism of TIMP3 gene expression and may provide evidence for the molecular diagnosis and stage evaluation of gastric cancer.

Virtual slides

The virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1756134016954958 webcite

【 授权许可】

   
2013 Guan et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Fang JY, Lu YY: Effects of histone acetylation and DNA methylation on p21(WAF1) regulation. World J Gastroenterol 2002, 8:400-405.
• [2]Kumar P, Xi Y: MicroRNA, epigenetic machinery and lung cancer. Thoracic Cancer 2011, 2:35-44.
• [3]Kimura N, Nagasaka T, Murakami J, Sasamoto H, Murakami M, Tanaka N, Matsubara N: Methylation profiles of genes utilizing newly developed CpG island methylation microarray on colorectal cancer patients. Nucleic Acids Res 2005, 33:e46.
• [4]Lee S, Hwang KS, Lee HJ, Kim JS, Kang GH: Aberrant CpG island hypermethylation of multiple genes in colorectal neoplasia. Lab Invest 2004, 84:884-893.
• [5]Kang GH, Lee HJ, Hwang KS, Lee S, Kim JH, Kim JS: Aberrant CpG island hyperme- thylation of chronic gastritis, in relation to aging, gender, intestinal metaplasia, and chronic inflammation. Am J Pathol 2003, 163:1551-1556.
• [6]Oue N, Matsumura S, Nakayama H, Kitadai Y, Taniyama K, Matsusaki K, Yasui W: Reduced expression of the TSP1 gene and its association with promoter hypermethylation in gastric carcinoma. Oncology 2003, 64:423-429.
• [7]Xue WC, Chan KY, Feng HC, Chiu PM, Ngan HY, Tsao SW, Cheung AN: Promoter hypermethylation of multiple genes in hydatidiform mole and choriocarcinoma. Mol Diagn 2004, 6:326-334.
• [8]Kim H, Kim YH, Kim SE, Kim NG, Noh SH, Kim H: Concerted promoter hypermethy- lation of hMLH1, p16INK4A, and E-cadherin in gastric carcinomas with microsatellite instability. J Pathol 2003, 200:23-31.
• [9]Lindsey JC, Lusher ME, Anderton JA, Bailey S, Gilbertson RJ, Pearson AD, Ellison DW, Clifford SC: Identification of tumour-specific epigenetic events in medullo- blastoma development by hypermethylation profiling. Carcinogenesis 2004, 25:661-668.
• [10]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.
• [11]Liu X, Xiong H, Li J, He Y, Yuan X: Correlation of hK6 expression with tumor recurrence and prognosis in advanced gastric cancer. Diagn Pathol 2013, 8:62. BioMed Central Full Text
• [12]Dong X, Wang G, Zhang G, Ni Z, Suo J, Cui J, et al.: The endothelial lipase protein is promising urinary biomarker for diagnosis of gastric cancer. Diagn Pathol 2013, 8:45. BioMed Central Full Text
• [13]Herman JG, Graff JR, Myohanen S, Nelkin BD, Baylin SB: Methylation-specific PCR: A novel PCR assay for methylation status of CpG island. Proc Natl Acad Sci USA 1996, 93:9821-9826.
• [14]Shimizu M, Saitoh Y, Itoh H: Immunohistochemical staining of Ha-ras oncogene product in normal, benign, and malignant human pancreatic tissue. Hum Pathol 1990, 21:607-612.
• [15]Vogelauer M, Wu J, Suka N, Grunstein M: Global histone acetylation and deacetylation in yeast. Nature 2000, 408:495-498.
• [16]Sadiq A, Mansour KA: Esophageal cancer: recent advances. Thoracic Cancer 2011, 2:75-83.
• [17]Suter CM, Norrie M, Ku SL, Cheong KF, Tomlinson I, Ward RL: CpG island methylation is a common finding in colorectal cancer cell lines. Br J Cancer 2003, 88(3):413-419.
• [18]Baylin SB, Herman JG: DNA hypermethylation in tumorigenesis: epigenetics joins genetics. Trends Genet 2000, 16:168-174.
• [19]Colot V, Maloisel L, Rossignol JL: DNA repeats and homologous recombination: a probable role for DNA methylation in genome stability of eukaryotic cells. J Soc Biol 1999, 193:29-34.
• [20]Kaneda A, Kaminishi M, Yanagihara K, Sugimura T, Ushijima T: Identification of silencing of nine genes in human gastric cancer. Cancer Res 2002, 62:6645-6650.
• [21]Leco KJ, Waterhouse P, Sanchez OH, Gowing KL, Poole AR, Wakeham A, Mak TW, Khokha R: Spontaneous air space enlargement in the lungs of mice lacking tissue inhibitor of metalloproteinases-3 (TIMP-3). J Clin Invest 2001, 108:817-829.
• [22]Mannello F, Gazzanelli G: Tissue inhibitors of metalloproteinases and programmed cell death: conundrums, controversies and potential implications. Apoptosis 2001, 6:479-482.
• [23]Smith MR, Kung H, Durum SK, Colburn NH, Sun Y: TIMP-3 induces cell death by stabilizing TNF-alpha receptors on the surface of human colon carcinoma cells. Cytokine 1997, 9:770-780.
• [24]Gagnon J, Shaker S, Primeau M, Hurtubise A, Momparler RL: Interaction of 5-aza-2′- deoxycytidine and depsipeptide on antineoplastic activity and activation of 14-3-3sigma, E-cadherin and tissue inhibitor of metalloproteinase 3 expression in human breast carcinoma cells. Anticancer Drugs 2003, 14:193-202.
• [25]Feng H, Cheung AN, Xue WC, Wang Y, Wang X, Fu S, Wang Q, Ngan HY, Tsao SW: Down-regulation and promoter methylation of tissue inhibitor of metalloproteinase 3 in choriocarcinoma. Gynecol Oncol 2004, 94:375-382.
• [26]Youn HG, An JY, Choi MG, Noh JH, Sohn TS, Kim S: Recurrence after curative resection of early gastric cancer. Ann Surg Oncol 2010, 17:448-454.
• [27]Jin J, Jin T, Quan M, Piao Y, Lin Z: Ezrin overexpression predicts the poor prognosis of gastric adenocarcinoma. Diagn Pathol 2012, 7:135. BioMed Central Full Text
• [28]Fan LL, Chen DF, Lan CH, Liu KY, Fang DC: Knockdown of ezrin via RNA interference suppresses Helicobacter pylori-enhanced invasion of gastric cancer cells. Cancer Biol Ther 2011, 11:746-752.
• [29]Kerbel RS: Growth dominance of the metastatic cancer cell: cellular and molecular aspects. Adv Cancer Res 1990, 55:87-131.
• [30]Yegnasubramanian S, Kowalski J, Gonzalgo ML, Zahurak M, Piantadosi S, Walsh PC, Bova GS, De Marzo AM, Isaacs WB, Nelson WG: Hypermethylation of CpG islands in primary and metastatic human prostate cancer. Cancer Res 2004, 64:1975-1986.
• [31]Lee S, Lee HJ, Kim JH, Lee HS, Jang JJ, Kang GH: Aberrant CpG island hypermethylation along multistep hepatocarcinogenesis. Am J Pathol 2003, 163:1371-1378.
• [32]Galm O, Rountree MR, Bachman KE, Jair KW, Baylin SB, Herman JG: Enzymatic regional methylation assay:a novel method quantify regional CpG methylation densuty. Genome Res 2002, 12:153-157.
• [33]Kang GH, Lee S, Shim YH, Kim JC, Ro JY, Shim Y-H: Profile of methylated CpG sites of hMLH1 promoter in primary gastric carcinoma with microsatellite instability. Cancer Res 2001, 61:2847-2851.