The online version of this article (doi:10.1186/1476-4598-11-40) contains supplementary material, which is available to authorized users.
The authors declare that they have no competing interests.
AD carried out the experiments, analyzed the results and drafted the manuscript. HC planned and coordinated the study, performed experimental and statistical analysis and revised the manuscript. TM and PK provided clinical information. All authors read and approved the final manuscript.
Hypermethylation of promotor CpG islands is a common mechanism that inactivates tumor suppressor genes in cancer. Genes belonging to the RASSF gene family have frequently been reported as epigenetically silenced by promotor methylation in human cancers. Two members of this gene family, RASSF1A and RASSF5A have been reported as methylated in neuroblastoma. Data from our previously performed genome-wide DNA methylation array analysis indicated that other members of the RASSF gene family are targeted by DNA methylation in neuroblastoma.
In the current study, we found that several of the RASSF family genes (RASSF2, RASSF4, RASSF5, RASSF6, RASSF7, and RASSF10) to various degrees were methylated in neuroblastoma cell lines and primary tumors. In addition, several of the RASSF family genes showed low or absent mRNA expression in neuroblastoma cell lines. RASSF5 and RASSF6 were to various degrees methylated in a large portion of neuroblastoma tumors and RASSF7 was heavily methylated in most tumors. Further, CpG methylation sites in the CpG islands of some RASSF family members could be used to significantly discriminate between biological subgroups of neuroblastoma tumors. For example, RASSF5 methylation highly correlated to MYCN amplification and INRG stage M. Furthermore, high methylation of RASSF6 was correlated to unfavorable outcome, 1p deletion and MYCN amplification in our tumor material.
This study shows that several genes belonging to the RASSF gene family are methylated in neuroblastoma. The genes RASSF5, RASSF6 and RASSF7 stand out as the most promising candidate genes for further investigations in neuroblastoma.
Authors’ original file for figure 112943_2012_1032_MOESM1_ESM.pdf
Authors’ original file for figure 212943_2012_1032_MOESM2_ESM.pdf
Authors’ original file for figure 312943_2012_1032_MOESM3_ESM.pdf
Authors’ original file for figure 412943_2012_1032_MOESM4_ESM.pdf
Authors’ original file for figure 512943_2012_1032_MOESM5_ESM.pdf
Carén H, Kryh H, Nethander M, Sjöberg RM, Träger C, Nilsson S, Abrahamsson J, Kogner P, Martinsson T: High-risk neuroblastoma tumors with 11q-deletion display a poor prognostic, chromosome instability phenotype with later onset. Proc Natl Acad Sci U S A. 2010, 107 (9): 4323-4328. 10.1073/pnas.0910684107 PubMedCentralCrossRefPubMed
Hesson LB, Dunwell TL, Cooper WN, Catchpoole D, Brini AT, Chiaramonte R, Griffiths M, Chalmers AD, Maher ER, Latif F: The novel RASSF6 and RASSF10 candidate tumour suppressor genes are frequently epigenetically inactivated in childhood leukaemias. Mol Cancer. 2009, 8: 42- 10.1186/1476-4598-8-42 PubMedCentralCrossRefPubMed
Kryh H, Carén H, Erichsen J, Sjöberg RM, Abrahamsson J, Kogner P, Martinsson T: Comprehensive SNP array study of frequently used neuroblastoma cell lines; copy neutral loss of heterozygosity is common in the cell lines but uncommon in primary tumors. BMC Genomics. 2011, 12: 443- 10.1186/1471-2164-12-443 PubMedCentralCrossRefPubMed
Hesson LB, Wilson R, Morton D, Adams C, Walker M, Maher ER, Latif F: CpG island promoter hypermethylation of a novel Ras-effector gene RASSF2A is an early event in colon carcinogenesis and correlates inversely with K-ras mutations. Oncogene. 2005, 24 (24): 3987-3994. 10.1038/sj.onc.1208566 CrossRefPubMed
Tommasi S, Dammann R, Zhang Z, Wang Y, Liu L, Tsark WM, Wilczynski SP, Li J, You M, Pfeifer GP: Tumor susceptibility of Rassf1a knockout mice. Cancer Res. 2005, 65 (1): 92-98. PubMed
van der Weyden L, Tachibana KK, Gonzalez MA, Adams DJ, Ng BL, Petty R, Venkitaraman AR, Arends MJ, Bradley A: The RASSF1A isoform of RASSF1 promotes microtubule stability and suppresses tumorigenesis. Mol Cell Biol. 2005, 25 (18): 8356-8367. 10.1128/MCB.25.18.8356-8367.2005 PubMedCentralCrossRefPubMed
Cooper WN, Dickinson RE, Dallol A, Grigorieva EV, Pavlova TV, Hesson LB, Bieche I, Broggini M, Maher ER, Zabarovsky ER: Epigenetic regulation of the ras effector/tumour suppressor RASSF2 in breast and lung cancer. Oncogene. 2008, 27 (12): 1805-1811. 10.1038/sj.onc.1210805 PubMedCentralCrossRefPubMed
Steinmann K, Sandner A, Schagdarsurengin U, Dammann RH: Frequent promoter hypermethylation of tumor-related genes in head and neck squamous cell carcinoma. Oncol Rep. 2009, 22 (6): 1519-1526. PubMed
Macheiner D, Gauglhofer C, Rodgarkia-Dara C, Grusch M, Brachner A, Bichler C, Kandioler D, Sutterluty H, Mikulits W, Schulte-Hermann R: NORE1B is a putative tumor suppressor in hepatocarcinogenesis and may act via RASSF1A. Cancer Res. 2009, 69 (1): 235-242. 10.1158/0008-5472.CAN-08-2144 CrossRefPubMed
Wen Y, Wang Q, Zhou C, Yan D, Qiu G, Yang C, Tang H, Peng Z: Decreased Expression of RASSF6 Is a Novel Independent Prognostic Marker of a Worse Outcome in Gastric Cancer Patients after Curative Surgery. Ann Surg Oncol. 2011 Dec, 18 (13): 3858-67. 10.1245/s10434-011-1668-5. 10.1245/s10434-011-1668-5 CrossRefPubMed
Recino A, Sherwood V, Flaxman A, Cooper WN, Latif F, Ward A, Chalmers AD: Human RASSF7 regulates the microtubule cytoskeleton and is required for spindle formation, Aurora B activation and chromosomal congression during mitosis. Biochem J. 2010, 430 (2): 207-213. 10.1042/BJ20100883 PubMedCentralCrossRefPubMed
Lock FE, Underhill-Day N, Dunwell T, Matallanas D, Cooper W, Hesson L, Recino A, Ward A, Pavlova T, Zabarovsky E: The RASSF8 candidate tumor suppressor inhibits cell growth and regulates the Wnt and NF-kappaB signaling pathways. Oncogene. 2010, 29 (30): 4307-4316. 10.1038/onc.2010.192 CrossRefPubMed
Hill VK, Underhill-Day N, Krex D, Robel K, Sangan CB, Summersgill HR, Morris M, Gentle D, Chalmers AD, Maher ER: Epigenetic inactivation of the RASSF10 candidate tumor suppressor gene is a frequent and an early event in gliomagenesis. Oncogene. 2011, 30 (8): 978-989. 10.1038/onc.2010.471 CrossRefPubMed
- The RASSF gene family members RASSF5, RASSF6 and RASSF7 show frequent DNA methylation in neuroblastoma
- BioMed Central
Neu im Fachgebiet Onkologie
Mail Icon II