RASSF10 is an epigenetically silenced tumor suppressor in gastric cancer

Li,Zhenhua; Chang,Xiaojing; Dai,Dongqiu; Deng,Peng; Sun,Qiang

doi:10.3892/or.2014.3039

RASSF10 is an epigenetically silenced tumor suppressor in gastric cancer

Authors:
- Zhenhua Li
- Xiaojing Chang
- Dongqiu Dai
- Peng Deng
- Qiang Sun
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Affiliations: Department of Gastrointestinal Surgery, the Fourth Affiliated Hospital, China Medical University, Shenyang, Liaoning 110032, P.R. China
Published online on: February 20, 2014 https://doi.org/10.3892/or.2014.3039
Pages: 1661-1668

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Abstract

To better understand the role of the N-Terminal Ras association domain family (RASSF) genes in the development of gastric cancer, we examined the expression of RASSF7 and RASSF10 and RASSF10 methylation in gastric cancer. We found that RASSF10 expression was lost in six gastric cancer cell lines, and was rescued by a DNA demethylating agent and a histone deacetylase inhibitor. However, RASSF7 expression was strong in four cancer cell lines as well as in 87% of primary gastric cancer tissues. In contrast, RASSF7 expression was moderate in the GES-1 cell line and negative in 33.3% of the corresponding non-cancerous tissues. Analysis of RASSF10 methylation by methylation-specific PCR (MSP) and sequencing revealed that the methylation frequency in primary gastric carcinoma tissues was significantly higher compared to that in adjacent non-carcinoma tissues (61.6 vs. 38.4%; p<0.01). The methylation frequency in the tumor with invasion depth at T3 and T4 was significantly higher compared to that with invasion depth at T1 and T2 (67.1 vs. 37.5%; p<0.05). Hypermethylation of RASSF10 was found in the patients with lymph node metastasis, compared to those with unaffected lymph nodes (68.8 vs. 40.9%; p<0.05). Among the 4 gross types of the Borrmann classification, i.e. EGC, Borrmann Ⅰ, Borrmann Ⅱ, Borrmann Ⅲ and Borrmann Ⅳ, the last one was more frequently methylated (85.7 vs. 56.9%; p<0.05). The present study revealed that RASSF10 is an epigenetically silenced gene involved in tumor invasion and metastasis in gastric cancer, suggesting that the methylation status of RASSF10 may be a useful indicator to predict the malignant degree of gastric cancer.

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1	Jemal A, Bray F, Center MM, Farley J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar
2	Jones PA and Baylin SB: The fundamental role of epigenetic events in cancer. Nat Rev Genet. 3:415–428. 2002.PubMed/NCBI
3	Esteller M: Epigenetics in cancer. N Engl J Med. 358:1148–1159. 2008. View Article : Google Scholar
4	Jones PA and Baylin SB: The epigenomics of cancer. Cell. 128:683–692. 2007. View Article : Google Scholar : PubMed/NCBI
5	Fleisher AS, Esteller M, Wang S, et al: Hypermethylation of the hMLH1 gene promoter in human gastric cancers with microsatellite instability. Cancer Res. 59:1090–1095. 1999.
6	Lee YY, Kang SH, Seo JY, et al: Alterations of p16INK4A and p15INK4B genes in gastric carcinomas. Cancer. 80:1889–1896. 1997. View Article : Google Scholar : PubMed/NCBI
7	Oue N, Shigeishi H, Kuniyasu H, et al: promoter hypermethylation of MGMT is associated with protein loss in gastric carcinoma. Int J Cancer. 93:805–809. 2001. View Article : Google Scholar : PubMed/NCBI
8	Tsuchiya T, Tamura G, Sato K, et al: Distinct methylation patterns of two APC gene promoters in normal and cancerous gastric epithelia. Oncogene. 19:3642–3646. 2000.PubMed/NCBI
9	Byun DS, Lee MG, Chae KS, Ryu BG and Chi SG: Frequent epigenetic inactivation of RASSF1A by aberrant promoter hypermethylation in human gastric adenocarcinoma. Cancer Res. 61:7034–7038. 2001.PubMed/NCBI
10	Richter AM, Pfeifer GP and Dammann RH: The RASSF proteins in cancer; from epigenetic silencing to functional characterization. Biochim Biophys Acta. 1796:114–128. 2009.PubMed/NCBI
11	Sherwood V, Manbodh R, Sheppard C and Chalmers AD: RASSF7 is a member of a new family of RAS association domain-containing proteins and is required for completing mitosis. Mol Biol Cell. 19:1772–1782. 2008. View Article : Google Scholar : PubMed/NCBI
12	Sherwood V, Recino A, Jeffries A, Ward A and Chalmers AD: The N-terminal RASSF family: a new group of Ras-association-domain-containing proteins, with emerging links to cancer formation. Biochem J. 425:303–311. 2009. View Article : Google Scholar : PubMed/NCBI
13	Recino A, Sherwood V, Flaxman A, et al: Human RASSF7 regulates the microtubule cytoskeleton and is required for spindle formation, Aurora B activation and chromosomal congression during mitosis. Biochem J. 430:207–213. 2010. View Article : Google Scholar : PubMed/NCBI
14	Mutter GL, Baak JP, Fitzgerald JT, et al: Global expression changes of constitutive and hormonally regulated genes during endometrial neoplastic transformation. Gynecol Oncol. 83:177–185. 2001. View Article : Google Scholar
15	Logsdon CD, Simeone DM, Binkley C, et al: Molecular profiling of pancreatic adenocancer and chronic pancreatitis identifies multiple genes differentially regulated in pancreatic cancer. Cancer Res. 63:2649–2657. 2003.
16	Lowe AW, Olsen M, Hao Y, et al: Gene expression patterns in pancreatic tumors, cells and tissues. PLoS One. 2:e3232007. View Article : Google Scholar : PubMed/NCBI
17	Camps C, Buffa FM, Colella S, et al: hsa-miR-210 is induced by hypoxia and is an independent prognostic factor in breast cancer. Clin Cancer Res. 14:1340–1348. 2008. View Article : Google Scholar : PubMed/NCBI
18	Liang GP, Su YY, Chen J, Yang ZC, Liu YS and Luo XD: Analysis of the early adaptive response of endothelial cells to hypoxia via a long serial analysis of gene expression. Biochem Biophys Res Commun. 384:415–419. 2009. View Article : Google Scholar : PubMed/NCBI
19	Takahashi S, Ebihara A, Kajiho H, Kontani K, Nishina H and Katada T: RASSF7 negatively regulates pro-apoptotic JNK signaling by inhibiting the activity of phosphorylated-MKK7. Cell Death Differ. 18:645–655. 2011. View Article : Google Scholar : PubMed/NCBI
20	Falvella FS, Manenti G, Spinola M, et al: Identification of RASSF8 as a candidate lung tumor suppressor gene. Oncogene. 25:3934–3938. 2006. View Article : Google Scholar : PubMed/NCBI
21	Korkola JE, Houldsworth J, Chadalavada RS, et al: Down-regulation of stem cell genes, including those in a 200-kb gene cluster at 12p13.31, is associated with in vivo differentiation of human male germ cell tumors. Cancer Res. 66:820–827. 2006. View Article : Google Scholar : PubMed/NCBI
22	Lock FE, Underhill-Day N, Dunwell T, et al: The RASSF8 candidate tumor suppressor inhibits cell growth and regulates the Wnt and NF-κB signaling pathways. Oncogene. 29:4307–4316. 2010.PubMed/NCBI
23	Hitomi J, Christofferson DE, Ng A, et al: Identification of a molecular signaling network that regulates a cellular necrotic cell death pathway. Cell. 135:1311–1323. 2008. View Article : Google Scholar : PubMed/NCBI
24	Hesson LB, Dunwell TL, Cooper WN, et al: The novel RASSF6 and RASSF10 candidate tumour suppressor genes are frequently epigenetically inactivated in childhood leukaemias. Mol Cancer. 8:422009. View Article : Google Scholar : PubMed/NCBI
25	Schagdarsurengin U, Richter AM, Wöhler C and Dammann RH: Frequent epigenetic inactivation of RASSF10 in thyroid cancer. Epigenetics. 4:571–576. 2009. View Article : Google Scholar : PubMed/NCBI
26	Hill VK, Underhill-Day N, Krex D, et al: Epigenetic inactivation of the RASSF10 candidate tumor suppressor gene is a frequent and an early event in gliomagenesis. Oncogene. 30:978–989. 2011. View Article : Google Scholar : PubMed/NCBI
27	Helmbold P, Richter AM, Walesch S, et al: RASSF10 promoter hypermethylation is frequent in malignant melanoma of the skin but uncommon in nevus cell nevi. J Invest Dermatol. 132:687–694. 2012. View Article : Google Scholar : PubMed/NCBI
28	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001.
29	Choi IS and Wu TT: Epigenetic alterations in gastric carcinogenesis. Cell Res. 15:247–254. 2005. View Article : Google Scholar : PubMed/NCBI
30	Kang GH, Lee S, Kim JS and Jung HY: Profile of aberrant CpG island methylation along multistep gastric carcinogenesis. Lab Invest. 83:519–526. 2003. View Article : Google Scholar : PubMed/NCBI
31	Sun Y, Deng D, You WC, et al: Methylation of p16 CpG islands associated with malignant transformation of gastric dysplasia in a population-based study. Clin Cancer Res. 10:5087–5093. 2004. View Article : Google Scholar : PubMed/NCBI