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:

Epigenetic inactivation of paired box gene 5, a novel tumor suppressor gene, through direct upregulation of p53 is associated with prognosis in gastric cancer patients

A Corrigendum to this article was published on 19 July 2012

Abstract

Using genome-wide methylation screening, we identified that paired box gene 5 (PAX5) is involved in human cancer development. However, the function of PAX5 in gastric cancer (GC) development is largely unclear. We analyzed its epigenetic inactivation, biological functions and clinical application in GC. PAX5 was silenced in seven out of eight GC cell lines. A significant downregulation was also detected in paired gastric tumors compared with adjacent non-cancerous tissues. The downregulation of PAX5 was closely linked to the promoter hypermethylation status and could be restored with demethylation treatment. Ectopic expression of PAX5 in silenced GC cell lines (AGS and BGC823) inhibited colony formation and cell viability, arrested cell cycle, induced apoptosis, suppressed cell migration and invasion and repressed tumorigenicity in nude mice. Consistent with the induction of apoptosis by PAX5 in vitro, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) staining showed significantly enhanced apoptotic cells in PAX5-expressed tumors compared with the vector control tumors. On the other hand, knockdown of PAX5 by PAX5–short hairpin RNA increased the cell viability and proliferation. The anti-tumorigenic function of PAX5 was revealed to be mediated by upregulating downstream targets of tumor protein 53 (p53), p21, BCL2-associated X protein, metastasis suppressor 1 and tissue inhibitors of metalloproteinase 1, and downregulating BCL2, cyclin D1, mesenchymal–epithelial transition factor (MET) and matrix metalloproteinase 1. Immunoprecipitation assay demonstrated that PAX5 directly bound to the promoters of p53 and MET. Moreover, PAX5 hypermethylation was detected in 77% (144 of 187) of primary GCs compared with 10.5% (2/19) of normal gastric tissues (P<0.0001). GC patients with PAX5 methylation had a significant poor survival compared with the unmethylated cases as demonstrated by Cox regression model and log-rank test. In conclusion, PAX5 is a novel functional tumor suppressor in gastric carcinogenesis. Detection of methylated PAX5 can be utilized as an independent prognostic factor in GC.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

Abbreviations

5-Aza:

5-aza-2′-deoxycytidine

BAX:

BCL2-associated X protein

BGS:

bisulfite genomic sequencing

ChIP:

chromatin immunoprecipitation

CI:

confidence interval

GC:

gastric cancer

H. pylori :

Helicobacter pylori

MET:

mesenchymal epithelial transition factor

MMP1:

matrix metalloproteinase 1

MSP:

methylation specific PCR

MTSS1:

metastasis suppressor 1

ORF:

open reading frame

p53:

tumor protein 53

PAX5 :

paired box gene 5

PBS:

phosphate buffered saline

PI:

propidium iodide

qPCR:

quantitative polymerase chain reaction

RR:

relative risk

TIMP1:

tissue inhibitors of metalloproteinase 1

TUNEL:

terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labelling

References

  • Adams B, Dorfler P, Aguzzi A, Kozmik Z, Urbanek P, Maurer-Fogy I et al. (1992). Pax-5 encodes the transcription factor BSAP and is expressed in B lymphocytes, the developing CNS, and adult testis. Genes Dev 6: 1589–1607.

    Article  CAS  Google Scholar 

  • Barberis A, Widenhorn K, Vitelli L, Busslinger M . (1990). A novel B-cell lineage-specific transcription factor present at early but not late stages of differentiation. Genes Dev 4: 849–859.

    Article  CAS  Google Scholar 

  • Baumann Kubetzko FB, Di Paolo C, Maag C, Meier R, Schafer BW, Betts DR et al. (2004). The PAX5 oncogene is expressed in N-type neuroblastoma cells and increases tumorigenicity of a S-type cell line. Carcinogenesis 25: 1839–1846.

    Article  CAS  Google Scholar 

  • Benvenuti S, Comoglio PM . (2007). The MET receptor tyrosine kinase in invasion and metastasis. J Cell Physiol 213: 316–325.

    Article  CAS  Google Scholar 

  • Bottaro DP, Rubin JS, Faletto DL, Chan AM, Kmiecik TE, Vande Woude GF et al. (1991). Identification of the hepatocyte growth factor receptor as the c-met proto-oncogene product. Science 251: 802–804.

    Article  CAS  Google Scholar 

  • Busslinger M, Klix N, Pfeffer P, Graninger PG, Kozmik Z . (1996). Deregulation of PAX-5 by translocation of the Emu enhancer of the IgH locus adjacent to two alternative PAX-5 promoters in a diffuse large-cell lymphoma. Proc Natl Acad Sci USA 93: 6129–6134.

    Article  CAS  Google Scholar 

  • Carotta S, Holmes ML, Pridans C, Nutt SL . (2006). Pax5 maintains cellular identity by repressing gene expression throughout B cell differentiation. Cell Cycle 5: 2452–2456.

    Article  CAS  Google Scholar 

  • Chambers AF, Matrisian LM . (1997). Changing views of the role of matrix metalloproteinases in metastasis. J Natl Cancer Inst 89: 1260–1270.

    Article  CAS  Google Scholar 

  • Cobaleda C, Schebesta A, Delogu A, Busslinger M . (2007). Pax5: the guardian of B cell identity and function. Nat Immunol 8: 463–470.

    Article  CAS  Google Scholar 

  • el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM et al. (1993). WAF1, a potential mediator of p53 tumor suppression. Cell 75: 817–825.

    Article  CAS  Google Scholar 

  • Giordano S, Zhen Z, Medico E, Gaudino G, Galimi F, Comoglio PM . (1993). Transfer of motogenic and invasive response to scatter factor/hepatocyte growth factor by transfection of human MET protooncogene. Proc Natl Acad Sci USA 90: 649–653.

    Article  CAS  Google Scholar 

  • Haldar S, Negrini M, Monne M, Sabbioni S, Croce CM . (1994). Down-regulation of bcl-2 by p53 in breast cancer cells. Cancer Res 54: 2095–2097.

    CAS  PubMed  Google Scholar 

  • He G, Siddik ZH, Huang Z, Wang R, Koomen J, Kobayashi R et al. (2005). Induction of p21 by p53 following DNA damage inhibits both Cdk4 and Cdk2 activities. Oncogene 24: 2929–2943.

    Article  CAS  Google Scholar 

  • Hockenbery DM, Zutter M, Hickey W, Nahm M, Korsmeyer SJ . (1991). BCL2 protein is topographically restricted in tissues characterized by apoptotic cell death. Proc Natl Acad Sci USA 88: 6961–6965.

    Article  CAS  Google Scholar 

  • Holmes ML, Carotta S, Corcoran LM, Nutt SL . (2006). Repression of Flt3 by Pax5 is crucial for B-cell lineage commitment. Genes Dev 20: 933–938.

    Article  CAS  Google Scholar 

  • Jones PL, Veenstra GJ, Wade PA, Vermaak D, Kass SU, Landsberger N et al. (1998). Methylated DNA and MeCP2 recruit histone deacetylase to repress transcription. Nat Genet 19: 187–191.

    Article  CAS  Google Scholar 

  • Kang GH, Lee HJ, Hwang KS, Lee S, Kim JH, Kim JS . (2003a). Aberrant CpG island hypermethylation of chronic gastritis, in relation to aging, gender, intestinal metaplasia, and chronic inflammation. Am J Pathol 163: 1551–1556.

    Article  CAS  Google Scholar 

  • Kang GH, Lee S, Kim JS, Jung HY . (2003b). Profile of aberrant CpG island methylation along multistep gastric carcinogenesis. Lab Invest 83: 519–526.

    Article  CAS  Google Scholar 

  • Kanteti R, Nallasura V, Loganathan S, Tretiakova M, Kroll T, Krishnaswamy S et al. (2009). PAX5 is expressed in small-cell lung cancer and positively regulates c-Met transcription. Lab Invest 89: 301–314.

    Article  CAS  Google Scholar 

  • Kishimoto Y, Murakami Y, Shiraishi M, Hayashi K, Sekiya T . (1992). Aberrations of the p53 tumor suppressor gene in human non-small cell carcinomas of the lung. Cancer Res 52: 4799–4804.

    CAS  PubMed  Google Scholar 

  • Kovac CR, Emelyanov A, Singh M, Ashouian N, Birshtein BK . (2000). BSAP (Pax5)-importin alpha 1 (Rch1) interaction identifies a nuclear localization sequence. J Biol Chem 275: 16752–16757.

    Article  CAS  Google Scholar 

  • Kozmik Z, Wang S, Dorfler P, Adams B, Busslinger M . (1992). The promoter of the CD19 gene is a target for the B-cell-specific transcription factor BSAP. Mol Cell Biol 12: 2662–2672.

    Article  CAS  Google Scholar 

  • Lazzi S, Bellan C, Onnis A, De Falco G, Sayed S, Kostopoulos I et al. (2009). Rare lymphoid neoplasms coexpressing B- and T-cell antigens. The role of PAX-5 gene methylation in their pathogenesis. Hum Pathol 40: 1252–1261.

    Article  CAS  Google Scholar 

  • Lee BL, Kim WH, Jung J, Cho SJ, Park JW, Kim J et al. (2008). A hypoxia-independent up-regulation of hypoxia-inducible factor-1 by AKT contributes to angiogenesis in human gastric cancer. Carcinogenesis 29: 44–51.

    Article  CAS  Google Scholar 

  • Lee JH, Park SJ, Abraham SC, Seo JS, Nam JH, Choi C et al. (2004). Frequent CpG island methylation in precursor lesions and early gastric adenocarcinomas. Oncogene 23: 4646–4654.

    Article  CAS  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  Google Scholar 

  • Levine AJ . (1989). The p53 tumor suppressor gene and gene product. Princess Takamatsu Symp 20: 221–230.

    CAS  PubMed  Google Scholar 

  • Liu W, Li X, Chu E, Go M, Xu L, Zhao G et al. (2011). Paired box gene 5 is a novel tumor suppressor in hepatocellular carcinoma through interaction with p53 signaling pathway. Hepatology 53: 843–853.

    Article  CAS  Google Scholar 

  • Malkin D, Sexsmith E, Yeger H, Williams BR, Coppes MJ . (1994). Mutations of the p53 tumor suppressor gene occur infrequently in Wilms’ tumor. Cancer Res 54: 2077–2079.

    CAS  PubMed  Google Scholar 

  • Meza JL, Anderson J, Pappo AS, Meyer WH . (2006). Analysis of prognostic factors in patients with nonmetastatic rhabdomyosarcoma treated on intergroup rhabdomyosarcoma studies III and IV: the Children's Oncology Group. J Clin Oncol 24: 3844–3851.

    Article  Google Scholar 

  • Mhawech-Fauceglia P, Saxena R, Zhang S, Terracciano L, Sauter G, Chadhuri A et al. (2007). Pax-5 immunoexpression in various types of benign and malignant tumours: a high-throughput tissue microarray analysis. J Clin Pathol 60: 709–714.

    Article  CAS  Google Scholar 

  • Miyashita T, Reed JC . (1995). Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 80: 293–299.

    Article  CAS  Google Scholar 

  • Musgrove EA, Lee CS, Buckley MF, Sutherland RL . (1994). Cyclin D1 induction in breast cancer cells shortens G1 and is sufficient for cells arrested in G1 to complete the cell cycle. Proc Natl Acad Sci USA 91: 8022–8026.

    Article  CAS  Google Scholar 

  • Nutt SL, Urbanek P, Rolink A, Busslinger M . (1997). Essential functions of Pax5 (BSAP) in pro-B cell development: difference between fetal and adult B lymphopoiesis and reduced V-to-DJ recombination at the IgH locus. Genes Dev 11: 476–491.

    Article  CAS  Google Scholar 

  • Oltvai ZN, Milliman CL, Korsmeyer SJ . (1993). Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell 74: 609–619.

    Article  CAS  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  Google Scholar 

  • Quelle DE, Ashmun RA, Shurtleff SA, Kato JY, Bar-Sagi D, Roussel MF et al. (1993). Overexpression of mouse D-type cyclins accelerates G1 phase in rodent fibroblasts. Genes Dev 7: 1559–1571.

    Article  CAS  Google Scholar 

  • Rocha S, Martin AM, Meek DW, Perkins ND . (2003). p53 represses cyclin D1 transcription through down regulation of Bcl-3 and inducing increased association of the p52 NF-kappaB subunit with histone deacetylase 1. Mol Cell Biol 23: 4713–4727.

    Article  CAS  Google Scholar 

  • Ruvolo PP, Deng X, May WS . (2001). Phosphorylation of Bcl2 and regulation of apoptosis. Leukemia 15: 515–522.

    Article  CAS  Google Scholar 

  • Sauter W, Rosenberger A, Beckmann L, Kropp S, Mittelstrass K, Timofeeva M et al. (2008). Matrix metalloproteinase 1 (MMP1) is associated with early-onset lung cancer. Cancer Epidemiol Biomarkers Prev 17: 1127–1135.

    Article  CAS  Google Scholar 

  • Souabni A, Cobaleda C, Schebesta M, Busslinger M . (2002). Pax5 promotes B lymphopoiesis and blocks T cell development by repressing Notch1. Immunity 17: 781–793.

    Article  CAS  Google Scholar 

  • Stapleton P, Weith A, Urbanek P, Kozmik Z, Busslinger M . (1993). Chromosomal localization of seven PAX genes and cloning of a novel family member, PAX-9. Nat Genet 3: 292–298.

    Article  CAS  Google Scholar 

  • Stuart ET, Haffner R, Oren M, Gruss P . (1995). Loss of p53 function through PAX-mediated transcriptional repression. EMBO J 14: 5638–5645.

    Article  CAS  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  Google Scholar 

  • Visse R, Nagase H . (2003). Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry. Circ Res 92: 827–839.

    Article  CAS  Google Scholar 

  • Wang Y, Liu J, Smith E, Zhou K, Liao J, Yang GY et al. (2007). Downregulation of missing in metastasis gene (MIM) is associated with the progression of bladder transitional carcinomas. Cancer Invest 25: 79–86.

    Article  Google Scholar 

  • Ying J, Li H, Seng TJ, Langford C, Srivastava G, Tsao SW et al. (2006). Functional epigenetics identifies a protocadherin PCDH10 as a candidate tumor suppressor for nasopharyngeal, esophageal and multiple other carcinomas with frequent methylation. Oncogene 25: 1070–1080.

    Article  CAS  Google Scholar 

  • Yu J, Chu ES, Wang R, Wang S, Wu CW, Wong VW et al. (2010). Heme oxygenase-1 protects against steatohepatitis in both cultured hepatocytes and mice. Gastroenterology 138: 694–704.

    Article  CAS  Google Scholar 

  • Zwollo P, Desiderio S . (1994). Specific recognition of the blk promoter by the B-lymphoid transcription factor B-cell-specific activator protein. J Biol Chem 269: 15310–15317.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported by research grants of National Basic Research Program of China (973 Program, 2010CB529305), Hong Kong General Research Fund (473008), RFCID (11100022, 10090942), CUHK Group Research Scheme (3110043) and Scheme C.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to J Yu.

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

Li, X., Cheung, K., Ma, X. et al. Epigenetic inactivation of paired box gene 5, a novel tumor suppressor gene, through direct upregulation of p53 is associated with prognosis in gastric cancer patients. Oncogene 31, 3419–3430 (2012). https://doi.org/10.1038/onc.2011.511

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2011.511

Keywords

This article is cited by

Search

Quick links