Clinical and Experimental Medicine

, Volume 17, Issue 1, pp 121–129 | Cite as

CDKN1A histone acetylation and gene expression relationship in gastric adenocarcinomas

  • Fernanda WisnieskiEmail author
  • Danielle Queiroz Calcagno
  • Mariana Ferreira Leal
  • Leonardo Caires Santos
  • Carolina Oliveira Gigek
  • Elizabeth Suchi Chen
  • Sâmia Demachki
  • Ricardo Artigiani
  • Paulo Pimentel Assumpção
  • Laércio Gomes Lourenço
  • Rommel Rodríguez Burbano
  • Marília Cardoso Smith
Letter to the Editor


CDKN1A is a tumor suppressor gene involved in gastric carcinogenesis and is a potential target for histone deacetylase inhibitor-based therapies. Upregulation of CDKN1A is generally observed in several cell lines after histone deacetylase inhibitor treatment; however, little is known about the histone acetylation status associated with this gene in clinical samples, including gastric tumor tissue samples. Therefore, our goal was to quantify the H3K9 and H4K16 acetylation levels associated with three CDKN1A regions in 21 matched pairs of gastric adenocarcinoma and corresponding adjacent non-tumor samples by chromatin immunoprecipitation and to correlate these data with the gene expression. Our results demonstrated that the −402, −20, and +182 CDKN1A regions showed a significantly increased acetylation level in at least one of the histones evaluated (p < 0.05, for all comparisons), and these levels were positively correlated in gastric tumors. However, an inverse correlation was detected between both H3K9 and H4K16 acetylation at the −402 CDKN1A region and mRNA levels in gastric tumors (r = −0.51, p = 0.02; r = −0.60, p < 0.01, respectively). Furthermore, increased H4K16 acetylation at the −20 CDKN1A region was associated with gastric tumors of patients without lymph node metastasis (p = 0.04). These results highlight the complexity of these processes in gastric adenocarcinoma and contribute to a better understanding of CDKN1A regulation in carcinogenesis.


CDKN1A Histone acetylation Gene expression regulation Gastric cancer 



This study was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP; to MCS, FW, DQC, and MFL), the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; to MCS and RRB), and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES; to COG).

Compliance with ethical standards

Conflict of interest



  1. 1.
    Stivala LA, Cazzalini O, Prosperi E. The cyclin-dependent kinase inhibitor p21CDKN1A as a target of anti-cancer drugs. Curr Cancer Drug Targets. 2012;12:85–96.CrossRefPubMedGoogle Scholar
  2. 2.
    Dutto I, Tillhon M, Cazzalini O, Stivala LA, Prosperi E. Biology of the cell cycle inhibitor p21(CDKN1A): molecular mechanisms and relevance in chemical toxicology. Arch Toxicol. 2015;89:155–78.CrossRefPubMedGoogle Scholar
  3. 3.
    Do Nascimento Borges B, Burbano RM, Harada ML. Absence of CIP1/KIP1 hypermethylation in gastric cancer patients from Northern Brazil. In Vivo. 2010;24:579–82.PubMedGoogle Scholar
  4. 4.
    Wisnieski F, Calcagno DQ, Leal MF, Chen ES, Gigek CO, Santos LC, Pontes TB, Rasmussen LT, Payao SL, Assumpcao PP, Lourenco LG, Demachki S, Artigiani R, Burbano RR, Smith MC. Differential expression of histone deacetylase and acetyltransferase genes in gastric cancer and their modulation by trichostatin A. Tumour Biol. 2014;35:6373–81.CrossRefPubMedGoogle Scholar
  5. 5.
    Struhl K. Histone acetylation and transcriptional regulatory mechanisms. Genes Dev. 1998;12:599–606.CrossRefPubMedGoogle Scholar
  6. 6.
    Sachweh MC, Drummond CJ, Higgins M, Campbell J, Lain S. Incompatible effects of p53 and HDAC inhibition on p21 expression and cell cycle progression. Cell Death Dis. 2013;4:e533.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Wisnieski F, Calcagno DQ, Leal MF, dos Santos LC, de Gigek CO, Chen ES, Pontes TB, Assumpcao PP, de Assumpcao MB, Demachki S, Burbano RR, de Smith MA. Reference genes for quantitative RT-PCR data in gastric tissues and cell lines. World J Gastroenterol. 2013;19:7121–8.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Huang HS, Matevossian A, Jiang Y, Akbarian S. Chromatin immunoprecipitation in postmortem brain. J Neurosci Methods. 2006;156:284–92.CrossRefPubMedGoogle Scholar
  9. 9.
    Kardassis D, Papakosta P, Pardali K, Moustakas A. c-Jun transactivates the promoter of the human p21(WAF1/Cip1) gene by acting as a superactivator of the ubiquitous transcription factor Sp1. J Biol Chem. 1999;274:29572–81.CrossRefPubMedGoogle Scholar
  10. 10.
    Gomes NP, Bjerke G, Llorente B, Szostek SA, Emerson BM, Espinosa JM. Gene-specific requirement for P-TEFb activity and RNA polymerase II phosphorylation within the p53 transcriptional program. Genes Dev. 2006;20:601–12.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Donner AJ, Szostek S, Hoover JM, Espinosa JM. CDK8 is a stimulus-specific positive coregulator of p53 target genes. Mol Cell. 2007;27:121–33.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Mitani Y, Oue N, Hamai Y, Aung PP, Matsumura S, Nakayama H, Kamata N, Yasui W. Histone H3 acetylation is associated with reduced p21(WAF1/CIP1) expression by gastric carcinoma. J Pathol. 2005;205:65–73.CrossRefPubMedGoogle Scholar
  13. 13.
    Verdone L, Caserta M, Di Mauro E. Role of histone acetylation in the control of gene expression. Biochem Cell Biol. 2005;83:344–53.CrossRefPubMedGoogle Scholar
  14. 14.
    Mulholland NM, Soeth E, Smith CL. Inhibition of MMTV transcription by HDAC inhibitors occurs independent of changes in chromatin remodeling and increased histone acetylation. Oncogene. 2003;22:4807–18.CrossRefPubMedGoogle Scholar
  15. 15.
    Deckert J, Struhl K. Histone acetylation at promoters is differentially affected by specific activators and repressors. Mol Cell Biol. 2001;21:2726–35.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Wang Z, Zang C, Cui K, Schones DE, Barski A, Peng W, Zhao K. Genome-wide mapping of HATs and HDACs reveals distinct functions in active and inactive genes. Cell. 2009;138:1019–31.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Eberharter A, Becker PB. Histone acetylation: a switch between repressive and permissive chromatin. Second in review series on chromatin dynamics. EMBO Rep. 2002;3:224–9.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Di Cerbo V, Schneider R. Cancers with wrong HATs: the impact of acetylation. Brief Funct Genomics. 2013;12:231–43.CrossRefPubMedGoogle Scholar
  19. 19.
    Love IM, Sekaric P, Shi D, Grossman SR, Androphy EJ. The histone acetyltransferase PCAF regulates p21 transcription through stress-induced acetylation of histone H3. Cell Cycle. 2012;11:2458–66.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Khayat AS, Guimaraes AC, Calcagno DQ, Seabra AD, Lima EM, Leal MF, Faria MH, Rabenhorst SH, Assumpcao PP, Demachki S, Smith MA, Burbano RR. Interrelationship between TP53 gene deletion, protein expression and chromosome 17 aneusomy in gastric adenocarcinoma. BMC Gastroenterol. 2009;9:55.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Calcagno DQ, Freitas VM, Leal MF, de Souza CR, Demachki S, Montenegro R, Assumpcao PP, Khayat AS, de Smith MA, dos Santos AK, Burbano RR. MYC, FBXW7 and TP53 copy number variation and expression in gastric cancer. BMC Gastroenterol. 2013;13:141.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Silva TC, Leal MF, Calcagno DQ, de Souza CR, Khayat AS, dos Santos NP, Montenegro RC, Rabenhorst SH, Nascimento MQ, Assumpcao PP, de Arruda Cardoso Smith M, Burbano RR. hTERT, MYC and TP53 deregulation in gastric preneoplastic lesions. BMC Gastroenterol. 2012;12:85.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Leal MF, Calcagno DQ, de Borges da Costa JF, Silva TC, Khayat AS, Chen ES, Assumpcao PP, de Arruda Cardoso Smith M, Burbano RR. MYC, TP53, and chromosome 17 copy-number alterations in multiple gastric cancer cell lines and in their parental primary tumors. J Biomed Biotechnol. 2011;2011:631268.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Leal MF, Calcagno DQ, Khayat AS, Silva TC, Muniz JA, Assumpcao PP, de Arruda Cardoso Smith M, Burbano RR. hTERT and TP53 deregulation in intestinal-type gastric carcinogenesis in non-human primates. Clin Exp Med. 2013;13:221–4.CrossRefPubMedGoogle Scholar
  25. 25.
    Abbas T, Dutta A. p21 in cancer: intricate networks and multiple activities. Nat Rev Cancer. 2009;9:400–14.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Leal MF, Martins do Nascimento JL, da Silva CE, Vita Lamarao MF, Calcagno DQ, Khayat AS, Assumpcao PP, Cabral IR, de Arruda Cardoso Smith M, Burbano RR. Establishment and conventional cytogenetic characterization of three gastric cancer cell lines. Cancer Genet Cytogenet. 2009;195:85–91.CrossRefPubMedGoogle Scholar
  27. 27.
    Calcagno DQ, Leal MF, Taken SS, Assumpcao PP, Demachki S, de Smith MA, Burbano RR. Aneuploidy of chromosome 8 and C-MYC amplification in individuals from northern Brazil with gastric adenocarcinoma. Anticancer Res. 2005;25:4069–74.PubMedGoogle Scholar
  28. 28.
    Burbano RR, Assumpcao PP, Leal MF, Calcagno DQ, Guimaraes AC, Khayat AS, Takeno SS, Chen ES, De Arruda Cardoso Smith M. C-MYC locus amplification as metastasis predictor in intestinal-type gastric adenocarcinomas: CGH study in Brazil. Anticancer Res. 2006;26:2909–14.PubMedGoogle Scholar
  29. 29.
    Assumpcao PP, Ishak G, Chen ES, Takeno SS, Leal MF, Guimaraes AC, Calcagno DQ, Khayat AS, Demachki S, de Smith MA, Burbano RR. Numerical aberrations of chromosome 8 detected by conventional cytogenetics and fluorescence in situ hybridization in individuals from northern Brazil with gastric adenocarcinoma. Cancer Genet Cytogenet. 2006;169:45–9.CrossRefPubMedGoogle Scholar
  30. 30.
    Calcagno DQ, Leal MF, Seabra AD, Khayat AS, Chen ES, Demachki S, Assumpcao PP, Faria MH, Rabenhorst SH, Ferreira MV, de Arruda Cardoso Smith M, Burbano RR. Interrelationship between chromosome 8 aneuploidy, C-MYC amplification and increased expression in individuals from northern Brazil with gastric adenocarcinoma. World J Gastroenterol. 2006;12:6207–11.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Calcagno DQ, Leal MF, Assumpcao PP, Smith MA, Burbano RR. MYC and gastric adenocarcinoma carcinogenesis. World J Gastroenterol. 2008;14:5962–8.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Costa Raiol LC, Figueira Silva EC, Mendes da Fonseca D, Leal MF, Guimaraes AC, Calcagno DQ, Khayat AS, Assumpcao PP, de Arruda Cardoso Smith M, Burbano RR. Interrelationship between MYC gene numerical aberrations and protein expression in individuals from northern Brazil with early gastric adenocarcinoma. Cancer Genet Cytogenet. 2008;181:31–5.CrossRefPubMedGoogle Scholar
  33. 33.
    Calcagno DQ, Guimaraes AC, Leal MF, Seabra AD, Khayat AS, Pontes TB, Assumpcao PP, De Arruda Cardoso Smith M, Burbano RR. MYC insertions in diffuse-type gastric adenocarcinoma. Anticancer Res. 2009;29:2479–83.PubMedGoogle Scholar
  34. 34.
    Calcagno DQ, Leal MF, Demachki S, Araujo MT, Freitas FW, Oliveira e Souza D, Assumpcao PP, Ishak G, de Arruda Cardoso Smith M, Burbano RR. MYC in gastric carcinoma and intestinal metaplasia of young adults. Cancer Genet Cytogenet. 2010;202:63–6.CrossRefPubMedGoogle Scholar
  35. 35.
    de Souza CR, Leal MF, Calcagno DQ, Costa Sozinho EK, do Borges BN, Montenegro RC, Dos Santos AK, Dos Santos SE, Ribeiro HF, Assumpcao PP, Smith M, Burbano RR. MYC deregulation in gastric cancer and its clinicopathological implications. PLoS ONE. 2013;8:e64420.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Ribeiro HF, Alcantara DF, Matos LA, Sousa JM, Leal MF, Smith MA, Burbano RR, Bahia MO. Cytogenetic characterization and evaluation of c-MYC gene amplification in PG100, a new Brazilian gastric cancer cell line. Braz J Med Biol Res. 2010;43:717–21.CrossRefPubMedGoogle Scholar
  37. 37.
    da Costa JF, Leal MF, Silva TC, Andrade Junior EF, Rezende AP, Muniz JA, Lacreta Junior AC, Assumpcao PP, Calcagno DQ, Demachki S, Rabenhorst SH, de Smith MA, Burbano RR. Experimental gastric carcinogenesis in Cebus apella nonhuman primates. PLoS ONE. 2011;6:e21988.CrossRefGoogle Scholar
  38. 38.
    Li H, Wu X. Histone deacetylase inhibitor, Trichostatin A, activates p21WAF1/CIP1 expression through downregulation of c-myc and release of the repression of c-myc from the promoter in human cervical cancer cells. Biochem Biophys Res Commun. 2004;324:860–7.CrossRefPubMedGoogle Scholar
  39. 39.
    Ellis DJ, Lawman ZK, Bonham K. Histone acetylation is not an accurate predictor of gene expression following treatment with histone deacetylase inhibitors. Biochem Biophys Res Commun. 2008;367:656–62.CrossRefPubMedGoogle Scholar
  40. 40.
    Child ES, Mann DJ. The intricacies of p21 phosphorylation: protein/protein interactions, subcellular localization and stability. Cell Cycle. 2006;5:1313–9.CrossRefPubMedGoogle Scholar
  41. 41.
    Bornstein G, Bloom J, Sitry-Shevah D, Nakayama K, Pagano M, Hershko A. Role of the SCFSkp2 ubiquitin ligase in the degradation of p21Cip1 in S phase. J Biol Chem. 2003;278:25752–7.CrossRefPubMedGoogle Scholar
  42. 42.
    Nakano K, Mizuno T, Sowa Y, Orita T, Yoshino T, Okuyama Y, Fujita T, Ohtani-Fujita N, Matsukawa Y, Tokino T, Yamagishi H, Oka T, Nomura H, Sakai T. Butyrate activates the WAF1/Cip1 gene promoter through Sp1 sites in a p53-negative human colon cancer cell line. J Biol Chem. 1997;272:22199–206.CrossRefPubMedGoogle Scholar
  43. 43.
    Lauren P. The two histological main types of gastric carcinoma: diffuse and so-called intestinal-type carcinoma. An attempt at a histo-clinical classification. Acta Pathol Microbiol Scand. 1965;64:31–49.PubMedGoogle Scholar
  44. 44.
    Washington K. 7th edition of the AJCC cancer staging manual: stomach. Ann Surg Oncol. 2010;17:3077–9.CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Fernanda Wisnieski
    • 1
    Email author
  • Danielle Queiroz Calcagno
    • 2
  • Mariana Ferreira Leal
    • 1
    • 3
  • Leonardo Caires Santos
    • 1
  • Carolina Oliveira Gigek
    • 1
  • Elizabeth Suchi Chen
    • 1
  • Sâmia Demachki
    • 2
  • Ricardo Artigiani
    • 4
  • Paulo Pimentel Assumpção
    • 2
  • Laércio Gomes Lourenço
    • 5
  • Rommel Rodríguez Burbano
    • 6
  • Marília Cardoso Smith
    • 1
  1. 1.Disciplina de Genética, Departamento de Morfologia e GenéticaUniversidade Federal de São PauloSão PauloBrazil
  2. 2.Núcleo de Pesquisas em Oncologia, Hospital João de Barros BarretoUniversidade Federal do ParáBelémBrazil
  3. 3.Departamento de Ortopedia e TraumatologiaUniversidade Federal de São PauloSão PauloBrazil
  4. 4.Departamento de PatologiaUniversidade Federal de São PauloSão PauloBrazil
  5. 5.Disciplina de Gastroenterologia Cirúrgica, Departamento de CirurgiaUniversidade Federal de São PauloSão PauloBrazil
  6. 6.Laboratório de Citogenética Humana, Instituto de Ciências BiológicasUniversidade Federal do ParáBelémBrazil

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