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Analysis of 5-aza-2'-deoxycytidine-induced Gene Expression in Lung Cancer Cell Lines  

김창수 (고신대학교 의과대학 흉부외과학교실)
이해영 (고신대학교 의과대학 흉부외과학교실)
김종인 (고신대학교 의과대학 흉부외과학교실)
장희경 (고신대학교 의과대학 병리학교실)
박종욱 (계명대학교 의과대학 면역학교실)
조성래 (고신대학교 의과대학 흉부외과학교실)
Publication Information
Journal of Chest Surgery / v.37, no.12, 2004 , pp. 967-977 More about this Journal
Abstract
Background: DNA methylation is one of the important gene expression mechanisms of the cell. When cytosine of CpG dinucleotide in promotor is hypomethylated, expression of some genes that is controlled by this promoter is altered. In this study, the author investigated the effect of DNA demethylating agent, 5-aza-2'-deoxycytidine (ADC), on the expressions of cancer antigen genes, MHC and B7 in 4 lung cancer cell lines, NCIH1703, NCIH522, MRC-5, and A549. Material and Method: After treatment of cell lines, NCIH1703, NCIH522, MRC-5 and A549 with ADC (1 uM) for 48 hours, RT-PCR was performed by using the primers of MAGE, GAGE, NY-ESO-1, PSMA, CEA, and SCC antigen gene. In order to find the optimal ADC treatment condition for induction of cancer antigen, we studied the effect of ADC treatment time and dose on the cancer antigen gene expression. To know the effect of ADC on the expression of MHC or B7 and cell growth, cells were treated with 1 uM of ADC for 72 hours for FACS analysis or cells were treated with 0.2, 1 or 5 uM of ADC for 96 hours for cell counting. Result: After treatment of ADC (1 uM) for 48 hours, the expressions of MAGE, GAGE, NY-ESO-1, and PSMA genes increased in some cell lines. Among 6 MAGE isotypes tested, and gene expression of MAGE-1, -2, -3, -4 and -6 could be induced by ADC treatment. However, CEA gene expression did not change and SCC gene expression was decreased by ADC treatment. Gene expression was generally induced 24 - 28 hours after ADC treatment and expression of MAGE, GAGE, and NY-ESO-1 was maintained at least 14 days after ADC ADC teatment, and expression of MAGE, GAGE, and NY-ESO-1 was maintained at least 14 days after ADC teatment in ADC-Free medium. Most gene expression could be induced at 0.2 uM of ADC, but gene expression increased dependently on ADC treatment dose. The expression of MHC and B7 was not increased by ADC treatment in all four cell lines, and the growth rate of 4 cell lines decreased significantly with the increase of ADC concentrations. Conclusion: Treatment of lung cancer cell lines with ADC increases the gene expression MAGE, GAGE and NY-ESO-1 that are capable of induction of cytotoxic T lymphocyte response. We suggest that treatment with 1 uM of ADC for 48 hours and then culturing in ADC-free medium is optimal condition for induction of cancer antigen. However, ADC has no effect on MHC and B7 induction, additional modification for increase of expression of MHC, B7 and cytokine will be needed for production of efficient cancer cell vaccine.
Keywords
Neopplasm marker; Gene expression; Concinoma; Lung neoplasms;
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1 Knuth A, Jager D, Jager E. Cancer immunotherapy in clinical oncology. Cancer Chemother Pharmacol 2000;46: Suppl: S46-51   DOI
2 Gong J, Nikrui N, Chen D, et al. Fusions of human ovarian carcinoma cells with autologous or allogeneic dendritic cells induce antitumor immunity. J Immunol 2000;165:1705-11   DOI   PUBMED
3 Levitsky HI, Lazenby A, Hayashi RJ, Pardoll DM. In vivo priming of two distinct antitumor effector populations: the role of MHC class I expression. J Exp Med 1994;179: 1215-24.   DOI   ScienceOn
4 Wachsman JT. DNA methylation and the association between genetic and epigenetic changes: relation to carcinogenesis. Mutat Res 1997;375:1-8.   DOI   PUBMED
5 Tureci O, Sahin U, Zwick C, Koslowski M, Seitz G, Pfreundschuh M. Identification of a meiosis-specific protein as a member of the class of cancer/testis antigens. Proc Natl Acad Sci USA 1998;95:5211-6   DOI   ScienceOn
6 Picardo AL, Torres AJ, Maestro M, et al. Quantitative analysis of carcinoembryonic antigen, squamous cell carcinoma antigen, CA 125, and CA 50 cytosolic content in non-small cell lung cancer. Cancer 1994;73:2305-11.   DOI   ScienceOn
7 van der Bruggen P, Bastin J, Gajewski T, et al. A peptide encoded by human gene MAGE-3 and presented by HLA-A2 induces cytolytic T lymphocytes that recognize tumor cells expressing MAGE-3. Eur J Immunol 1994;24:3038-43.   DOI   ScienceOn
8 Mendiratta SK, Quezada A, Matar M, et al. Intratumoral delivery of IL-12 gene by polyvinyl polymeric vector system to murine renal and colon carcinoma results in potent antitumor immunity. Gene Ther 1999;6:833-9.   DOI   ScienceOn
9 Coral S, Sigalotti L, Gasparollo A, et al. Prolonged upregulation of the expression of HLA class I antigens and costimulatory molecules on melanoma cells treated with 5-aza-2'- deoxycytidine (5-AZA-CdR). J Immunother 1999;22:16-24.   DOI   PUBMED
10 Celis E, Fikes J, Wentworth P, et al. Identification of potential CTL epitopes of tumor-associated antigen MAGE-1 for five common HLA-A alleles. Mol Immunol 1994;31:1423-30.   DOI   ScienceOn
11 De Backer O, Arden KC, Boretti M, et al. Characterization of the GAGE genes that are expressed in various human cancers and in normal testis. Cancer Res 1999;59:3157-65.   PUBMED
12 Timmerman JM, Levy R. Dendritic cell vaccines for cancer immunotherapy. Annu Rev Med 1999;50:507-29.   DOI   ScienceOn
13 Lee SG, Heo DS, Yoon SJ, et al. Effect of GM-CSF and IL-2 co-expression on the anti-tumor immune response. Anticancer Res 2000;20:2681-6.   PUBMED
14 Russo V, Traversari C, Verrecchia A, Mottolese M, Natali PG, Bordignon C. Expression of MAGE gene family in primary and metastatic human breast cancer: implications for tumor-specific immunotherapy. Int J Cancer 1995;64:216-21   DOI   ScienceOn
15 Russo V, Dalerba P, Ricci A, et al. MAGE BAGE and GAGE genes expression in fresh epithelial ovarian carcinomas. Int J Cancel 1996;67:457-60.   DOI   ScienceOn
16 Ockert D, Schmitz M, Hampl M, Rieber EP. Advances in cancer immunotherapy. Immunol Today 2000;20:63-5.   DOI   ScienceOn
17 Momparler RL, Bovenzi V. DNA methylation and cancer. J Cell Physiol 2000;183:145-54.   DOI   ScienceOn
18 Mori M, Inoue H, Mimori K, et al. Expression of MAGE genes in human colorectal carcinoma. Ann Surg 1996;224: 183-8.   DOI   ScienceOn
19 Gonzalgo ML, Jones PA. Mutagenic and epigenetic effects of DNA methylation. Mutat Res 1997;386:107-18.   DOI   ScienceOn
20 Zeuthen J, Dzhandzhugazyan K, Hansen MR, Kirkin AF. The immunogenic properties of human melanomas and melanoma-associated antigens recognized by cytotoxic T lymphocytes. Bratisl Lek Listy 1998;99:426-34   PUBMED
21 Schadendorf D, Paschen A, Sun Y. Autologous, allogeneic tumor cells or genetically engineered cells as cancer vaccine against melanoma. Immunol Lett 2000;74:67-74   DOI   ScienceOn
22 De Backer O, Arden KC, Boretti M, et al. Characterization of the GAGE genes that are expressed in various human cancers and in normal testis. Cancer Res 1999;59:3157-65.   PUBMED
23 Coral S, Sigalotti L, Gasparollo A, et al. Prolonged upregulation of the expression of HLA class I antigens and costimulatory molecules on melanoma cells treated with 5-aza- 2'-deoxycytidine (5-AZA-CdR). J Immunother 1999;22:16-24.   DOI   PUBMED
24 Chen JL, Dunbar PR, Gileadi U, et al. Identification of NY- ESO-1 peptide analogues capable of improved stimulation of tumor-reactive CTL. J Immunol 2000;165:948-55.   DOI   PUBMED
25 Chen ME, Lin SH, Chung LW, Sikes RA. Isolation and characterization of PAGE-1 and GAGE-7. New genes expressed in the LNCaP prostate cancer progression model that share homology with melanoma-associated antigens. J Biol Chem 1998;273:17618-25.   DOI   ScienceOn
26 Lee YC, Yang PC, Kuo SH, Luh KT. Tissue polypeptide antigen and carcinoembryonic antigen as tumor markers in lung cancer. J Formos Med Assoc 1991;90:631-6.   PUBMED
27 Fujie T, Mori M, Ueo H, Sugimachi K, Akiyoshi T. Expression of MAGE and BAGE genes in Japanese breast cancers. Ann Oncol 1997;8:369-72.   DOI   ScienceOn
28 Ockert D, Schmitz M, Hampl M, Rieber EP. Advances in cancer immunotherapy. Immunol Today 2000;20:63-5.   DOI   ScienceOn
29 Li J, Yang Y, Fujie T, et al. Expression of the MAGE gene family in human gastric carcinoma. Anticancer Res 1997; 17:3559-63   PUBMED
30 Redondo M, Ruiz-Cabello F, Concha A, et al. Differential expression of MHC class II genes in lung tumour cell lines. Eur J Immunogenet 25:385-91.