Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

MTSS1, a novel target of DNA methyltransferase 3B, functions as a tumor suppressor in hepatocellular carcinoma

Oncogene volume 31, pages 2298–2308 (2012)Cite this article

Abstract

DNA methyltransferase 3B (DNMT3B) mediates gene silencing via epigenetic mechanisms during hepatocellular carcinoma (HCC) progression. We aimed to identify novel targets of DNMT3B and their potential regulatory mechanisms in HCC. Metastasis suppressor 1 (MTSS1) was one of the DNMT3B targets and selected for further study. DNMT3B overexpression was detected in 81.25% of clinical HCC specimens and was negatively associated with MTSS1 in HCC cells and clinical samples. The underlying mechanism by which DNMT3B silences MTSS1 was studied using a combination of methylation-specific polymerase chain reaction (PCR) and bisulfite genome sequencing, chromatin immunoprecipitation-PCR and luciferase reporter assays. We found that the MTSS1 promoter region was sparsely methylated, and the methylation inhibitors failed to abolish DNMT3B-mediated MTSS1 silencing. DNMT3B protein bound directly to the 5′-flanking region (−865/−645) of the MTSS1 gene to inhibit its transcription. The functional role of MTSS1 was investigated using in vitro and in vivo tumorigenicity assays. As a result, MTSS1 exerted tumor suppressor effects and arrested cells in the G2/M phase, but not the G1/S phase of the cell cycle when it was depleted or overexpressed in HCC cells. Taken together, MTSS1, a novel target of DNMT3B, is repressed by DNMT3B via a DNA methylation-independent mechanism. MTSS1 was further characterized as a novel tumor suppressor gene in HCC. These findings highlight how DNMT3B regulates MTSS1, and such data may be useful for the development of new treatment options for HCC.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

Abbreviations

5-Aza:

5-aza-2′-deoxycytidine

BGS:

bisulfite sequencing

ChIP:

chromatin immunoprecipitation

DNMTs:

DNA methyltransferases

HCC:

hepatocellular carcinoma

MSP:

methylation-specific PCR

PI:

propidium iodide

qPCR:

quantitative real-time PCR

siRNA:

small interfering RNA

TSG:

tumor suppressor gene

References

  • Bachman KE, Rountree MR, Baylin SB . (2001). Dnmt3a and Dnmt3b are transcriptional repressors that exhibit unique localization properties to heterochromatin. J Biol Chem 276: 32282–32287.

    Article  CAS  PubMed  Google Scholar 

  • Baylin SB, Herman JG, Graff JR, Vertino PM, Issa JP . (1998). Alterations in DNA methylation: a fundamental aspect of neoplasia. Adv Cancer Res 72: 141–196.

    Article  CAS  PubMed  Google Scholar 

  • Beaulieu N, Morin S, Chute IC, Robert MF, Nguyen H, MacLeod AR . (2002). An essential role for DNA methyltransferase DNMT3B in cancer cell survival. J Biol Chem 277: 28176–28181.

    Article  CAS  PubMed  Google Scholar 

  • Bestor TH . (2000). The DNA methyltransferases of mammals. Hum Mol Genet 9: 2395–2402.

    Article  CAS  PubMed  Google Scholar 

  • Chang DC, Xu N, Luo KQ . (2003). Degradation of cyclin B is required for the onset of anaphase in mammalian cells. J Biol Chem 278: 37865–37873.

    Article  CAS  PubMed  Google Scholar 

  • Chang HC, Cho CY, Hung WC . (2006). Silencing of the metastasis suppressor RECK by RAS oncogene is mediated by DNA methyltransferase 3b-induced promoter methylation. Cancer Res 66: 8413–8420.

    Article  CAS  PubMed  Google Scholar 

  • Chen L, Chan TH, Yuan YF, Hu L, Huang J, Ma S et al. (2010). CHD1L promotes hepatocellular carcinoma progression and metastasis in mice and is associated with these processes in human patients. J Clin Invest 120: 1178–1191.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Choi MS, Shim YH, Hwa JY, Lee SK, Ro JY, Kim JS et al. (2003). Expression of DNA methyltransferases in multistep hepatocarcinogenesis. Hum Pathol 34: 11–17.

    Article  CAS  PubMed  Google Scholar 

  • Claus R, Lubbert M . (2003). Epigenetic targets in hematopoietic malignancies. Oncogene 22: 6489–6496.

    Article  CAS  PubMed  Google Scholar 

  • Cross FR, Schroeder L, Kruse M, Chen KC . (2005). Quantitative characterization of a mitotic cyclin threshold regulating exit from mitosis. Mol Biol Cell 16: 2129–2138.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Esteller M . (2008). Epigenetics in cancer. N Engl J Med 358: 1148–1159.

    Article  CAS  PubMed  Google Scholar 

  • Girault I, Tozlu S, Lidereau R, Bieche I . (2003). Expression analysis of DNA methyltransferases 1, 3A, and 3B in sporadic breast carcinomas. Clin Cancer Res 9: 4415–4422.

    CAS  PubMed  Google Scholar 

  • Gowher H, Jeltsch A . (2002). Molecular enzymology of the catalytic domains of the Dnmt3a and Dnmt3b DNA methyltransferases. J Biol Chem 277: 20409–20414.

    Article  CAS  PubMed  Google Scholar 

  • Huang XY, Huang ZL, Xu YH, Zhou J, Ye SL, Tang ZY et al. (2010). Effects of MIM-B gene on invasive and metastatic potentials of human hepatocelluar carcinoma MHCC97H cells. Zhonghua Gan Zang Bing Za Zhi 18: 915–919.

    CAS  PubMed  Google Scholar 

  • Jones PA, Takai D . (2001). The role of DNA methylation in mammalian epigenetics. Science 293: 1068–1070.

    Article  CAS  PubMed  Google Scholar 

  • Kanai Y, Hirohashi S . (2007). Alterations of DNA methylation associated with abnormalities of DNA methyltransferases in human cancers during transition from a precancerous to a malignant state. Carcinogenesis 28: 2434–2442.

    Article  CAS  PubMed  Google Scholar 

  • Kaneda M, Okano M, Hata K, Sado T, Tsujimoto N, Li E et al. (2004). Essential role for de novo DNA methyltransferase Dnmt3a in paternal and maternal imprinting. Nature 429: 900–903.

    Article  CAS  PubMed  Google Scholar 

  • Lee YG, Macoska JA, Korenchuk S, Pienta KJ . (2002). MIM, a potential metastasis suppressor gene in bladder cancer. Neoplasia 4: 291–294.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Linhart HG, Lin H, Yamada Y, Moran E, Steine EJ, Gokhale S et al. (2007). Dnmt3b promotes tumorigenesis in vivo by gene-specific de novo methylation and transcriptional silencing. Genes Dev 21: 3110–3122.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu K, Wang G, Ding H, Chen Y, Yu G, Wang J . (2010). Downregulation of metastasis suppressor 1(MTSS1) is associated with nodal metastasis and poor outcome in Chinese patients with gastric cancer. BMC Cancer 10: 428.

    Article  PubMed  PubMed Central  Google Scholar 

  • Ma S, Guan XY, Lee TK, Chan KW . (2007). Clinicopathological significance of missing in metastasis B expression in hepatocellular carcinoma. Hum Pathol 38: 1201–1206.

    Article  CAS  PubMed  Google Scholar 

  • McGarvey KM, Greene E, Fahrner JA, Jenuwein T, Baylin SB . (2007). DNA methylation and complete transcriptional silencing of cancer genes persist after depletion of EZH2. Cancer Res 67: 5097–5102.

    Article  CAS  PubMed  Google Scholar 

  • Mizuno S, Chijiwa T, Okamura T, Akashi K, Fukumaki Y, Niho Y et al. (2001). Expression of DNA methyltransferases DNMT1, 3A, and 3B in normal hematopoiesis and in acute and chronic myelogenous leukemia. Blood 97: 1172–1179.

    Article  CAS  PubMed  Google Scholar 

  • Nixdorf S, Grimm MO, Loberg R, Marreiros A, Russell PJ, Pienta KJ et al. (2004). Expression and regulation of MIM (Missing In Metastasis), a novel putative metastasis suppressor gene, and MIM-B, in bladder cancer cell lines. Cancer Lett 215: 209–220.

    Article  CAS  PubMed  Google Scholar 

  • Oh BK, Kim H, Park HJ, Shim YH, Choi J, Park C et al. (2007). DNA methyltransferase expression and DNA methylation in human hepatocellular carcinoma and their clinicopathological correlation. Int J Mol Med 20: 65–73.

    CAS  PubMed  Google Scholar 

  • Okano M, Bell DW, Haber DA, Li E . (1999). DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell 99: 247–257.

    Article  CAS  PubMed  Google Scholar 

  • Okano M, Xie S, Li E . (1998). Cloning and characterization of a family of novel mammalian DNA (cytosine-5) methyltransferases. Nat Genet 19: 219–220.

    Article  CAS  PubMed  Google Scholar 

  • Parr C, Jiang WG . (2009). Metastasis suppressor 1 (MTSS1) demonstrates prognostic value and anti-metastatic properties in breast cancer. Eur J Cancer 45: 1673–1683.

    Article  CAS  PubMed  Google Scholar 

  • Parry DH, O'Farrell PH . (2001). The schedule of destruction of three mitotic cyclins can dictate the timing of events during exit from mitosis. Curr Biol 11: 671–683.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Potapova TA, Daum JR, Pittman BD, Hudson JR, Jones TN, Satinover DL et al. (2006). The reversibility of mitotic exit in vertebrate cells. Nature 440: 954–958.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rhee I, Bachman KE, Park BH, Jair KW, Yen RW, Schuebel KE et al. (2002). DNMT1 and DNMT3b cooperate to silence genes in human cancer cells. Nature 416: 552–556.

    Article  CAS  PubMed  Google Scholar 

  • Robertson KD . (2001). DNA methylation, methyltransferases, and cancer. Oncogene 20: 3139–3155.

    Article  CAS  PubMed  Google Scholar 

  • Robertson KD, Ait-Si-Ali S, Yokochi T, Wade PA, Jones PL, Wolffe AP . (2000). DNMT1 forms a complex with Rb, E2F1 and HDAC1 and represses transcription from E2F-responsive promoters. Nat Genet 25: 338–342.

    Article  CAS  PubMed  Google Scholar 

  • Robertson KD, Jones PA . (2000). DNA methylation: past, present and future directions. Carcinogenesis 21: 461–467.

    Article  CAS  PubMed  Google Scholar 

  • Robertson KD, Keyomarsi K, Gonzales FA, Velicescu M, Jones PA . (2000). Differential mRNA expression of the human DNA methyltransferases (DNMTs) 1, 3a and 3b during the G(0)/G(1) to S phase transition in normal and tumor cells. Nucleic Acids Res 28: 2108–2113.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Roll JD, Rivenbark AG, Jones WD, Coleman WB . (2008). DNMT3b overexpression contributes to a hypermethylator phenotype in human breast cancer cell lines. Mol Cancer 7: 15.

    Article  PubMed  PubMed Central  Google Scholar 

  • Rountree MR, Bachman KE, Baylin SB . (2000). DNMT1 binds HDAC2 and a new co-repressor, DMAP1, to form a complex at replication foci. Nat Genet 25: 269–277.

    Article  CAS  PubMed  Google Scholar 

  • Saito Y, Kanai Y, Sakamoto M, Saito H, Ishii H, Hirohashi S . (2001). Expression of mRNA for DNA methyltransferases and methyl-CpG-binding proteins and DNA methylation status on CpG islands and pericentromeric satellite regions during human hepatocarcinogenesis. Hepatology 33: 561–568.

    Article  CAS  PubMed  Google Scholar 

  • Saito Y, Kanai Y, Sakamoto M, Saito H, Ishii H, Hirohashi S . (2002). Overexpression of a splice variant of DNA methyltransferase 3b, DNMT3b4, associated with DNA hypomethylation on pericentromeric satellite regions during human hepatocarcinogenesis. Proc Natl Acad Sci USA 99: 10060–10065.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shafiei F, Rahnama F, Pawella L, Mitchell MD, Gluckman PD, Lobie PE . (2008). DNMT3A and DNMT3B mediate autocrine hGH repression of plakoglobin gene transcription and consequent phenotypic conversion of mammary carcinoma cells. Oncogene 27: 2602–2612.

    Article  CAS  PubMed  Google Scholar 

  • Sigrist S, Jacobs H, Stratmann R, Lehner CF . (1995). Exit from mitosis is regulated by Drosophila fizzy and the sequential destruction of cyclins A, B and B3. EMBO J 14: 4827–4838.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Simao Tde A, Simoes GL, Ribeiro FS, Cidade DA, Andreollo NA, Lopes LR et al. (2006). Lower expression of p14ARF and p16INK4a correlates with higher DNMT3B expression in human oesophageal squamous cell carcinomas. Hum Exp Toxicol 25: 515–522.

    Article  CAS  Google Scholar 

  • Takeshima H, Suetake I, Shimahara H, Ura K, Tate S, Tajima S . (2006). Distinct DNA methylation activity of Dnmt3a and Dnmt3b towards naked and nucleosomal DNA. J Biochem 139: 503–515.

    Article  CAS  PubMed  Google Scholar 

  • Tao Q, Huang H, Geiman TM, Lim CY, Fu L, Qiu GH et al. (2002). Defective de novo methylation of viral and cellular DNA sequences in ICF syndrome cells. Hum Mol Genet 11: 2091–2102.

    Article  CAS  PubMed  Google Scholar 

  • Utikal J, Gratchev A, Muller-Molinet I, Oerther S, Kzhyshkowska J, Arens N et al. (2006). The expression of metastasis suppressor MIM/MTSS1 is regulated by DNA methylation. Int J Cancer 119: 2287–2293.

    Article  CAS  PubMed  Google Scholar 

  • Wang J, Bhutani M, Pathak AK, Lang W, Ren H, Jelinek J et al. (2007). Delta DNMT3B variants regulate DNA methylation in a promoter-specific manner. Cancer Res 67: 10647–10652.

    Article  CAS  PubMed  Google Scholar 

  • Weisenberger DJ, Velicescu M, Cheng JC, Gonzales FA, Liang G, Jones PA . (2004). Role of the DNA methyltransferase variant DNMT3b3 in DNA methylation. Mol Cancer Res 2: 62–72.

    CAS  PubMed  Google Scholar 

  • Xu J, Fan H, Zhao ZJ, Zhang JQ, Xie W . (2005). Identification of potential genes regulated by DNA methyltransferase 3B in a hepatocellular carcinoma cell line by RNA interference and microarray analysis. Yi Chuan Xue Bao 32: 1115–1127.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work was supported by The National Natural Science Foundation of China, No. 30971605, and in part by the Dr Cheng Yu Tung Fellowships 2007/08 under the HKU Li Ka Shing Faculty of Medicine. We are grateful to Professor Dianqing WU in UConn Health Center USA for providing siRNA expression vector. We are also grateful to Dr Stephanie Ma in Department of Pathology in University of Hong Kong for helping in MTSS1 antibody.

Author information

Author notes

  1. H Fan and L Chen: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medical Genetics and Developmental Biology, The Key Laboratory of Developmental Genes and Human Diseases, Ministry of Education, Medical School of Southeast University, Nanjing, China

    H Fan, F Zhang, Y Quan, X Su & X Qiu

  2. Department of Clinical Oncology, The University of Hong Kong, Pokfulam, Hong Kong, China

    L Chen, Z Zhao, K L Kong, S Dong, Y Song, T H M Chan & X-Y Guan

  3. State Key Laboratory for Liver Cancer, The University of Hong Kong, Pokfulam, Hong Kong, China

    L Chen, T H M Chan & X-Y Guan

  4. Department of Clinical Oncology, State Key Laboratory of Oncology in Southern China, Sun Yat-Sen University Cancer Center, Sun Yat-sen University, Guangzhou, China

    L Chen & X-Y Guan

Authors
  1. H Fan
    View author publications

    Search author on:PubMed Google Scholar

  2. L Chen
    View author publications

    Search author on:PubMed Google Scholar

  3. F Zhang
    View author publications

    Search author on:PubMed Google Scholar

  4. Y Quan
    View author publications

    Search author on:PubMed Google Scholar

  5. X Su
    View author publications

    Search author on:PubMed Google Scholar

  6. X Qiu
    View author publications

    Search author on:PubMed Google Scholar

  7. Z Zhao
    View author publications

    Search author on:PubMed Google Scholar

  8. K L Kong
    View author publications

    Search author on:PubMed Google Scholar

  9. S Dong
    View author publications

    Search author on:PubMed Google Scholar

  10. Y Song
    View author publications

    Search author on:PubMed Google Scholar

  11. T H M Chan
    View author publications

    Search author on:PubMed Google Scholar

  12. X-Y Guan
    View author publications

    Search author on:PubMed Google Scholar

Corresponding authors

Correspondence to H Fan or X-Y Guan.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies the paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fan, H., Chen, L., Zhang, F. et al. MTSS1, a novel target of DNA methyltransferase 3B, functions as a tumor suppressor in hepatocellular carcinoma. Oncogene 31, 2298–2308 (2012). https://doi.org/10.1038/onc.2011.411

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/onc.2011.411

Keywords

This article is cited by

Search

Advanced search

Quick links