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Ann Thorac Surg 2001;71:295-302
© 2001 The Society of Thoracic Surgeons

Original article: general thoracic

Induction of MAGE-3 expression in lung and esophageal cancer cells

Todd S. Weiser, MDa, Galen A. Ohnmacht, MDa, Zong S. Guo, PhDa, Maria R. Fischette, DOa, Gustavio A. Chen, MSa, Julie A. Hong, MSa, Dao M. Nguyen, MDa, David S. Schrump, MDa

a Thoracic Oncology Section, Surgery Branch, National Cancer Institute, Bethesda, Maryland, USA

Address reprint requests to Dr Schrump, Thoracic Oncology Section, Surgery Branch, National Cancer Institute, Building 10, Room 2B-07, 10 Center Dr, Bethesda, MD 20892-1502
e-mail: david_schrump{at}nih.gov

Presented at the Thirty-sixth Annual Meeting of The Society of Thoracic Surgeons, Fort Lauderdale, FL, Jan 31–Feb 2, 2000.

Background. Although MAGE-3 has been detected in approximately 40% of lung and esophageal cancers, expression of this cancer testis antigen appears to be below the threshold for immune recognition in patients with these malignancies. The aim of this study was to determine if the demethylating agent, 5-Aza-2'-deoxycytidine (DAC) and if the histone deacetylase inhibitor Depsipeptide FR901228 (DP) could enhance MAGE-3 expression in lung and esophageal cancer cells.

Methods. Eleven lung and esophageal cancer lines and cultured normal human bronchial epithelial (NHBE) cells were exposed to normal media (NM), DAC, DP, or combination DAC/DP at varying concentrations and exposure durations. MAGE-3 expression was evaluated by quantitative RT-PCR (TaqMan) and immunohistochemistry techniques. Trypan blue exclusion techniques were used to examine the proliferation of cancer cells after drug exposure.

Results. Relative to untreated controls, MAGE-3 expression was enhanced 32-fold (range 3.9 to 110) by DAC alone (0.1 µmol/L x 72 h), 2.1-fold (0.4 to 4.2) by DP alone (25 ng/mL x 6h), and 57-fold (4.6 to 209) by sequential DAC/DP exposure. Increased MAGE-3 mRNA copy numbers coincided with enhanced protein levels in these cells. MAGE-3 expression persisted after drug exposure. Flow cytometry confirmed the presence of functional HLA class I expression in these cells. Sequential DAC/DP treatment mediated pronounced growth inhibition in cancer cells but not NHBE.

Conclusions. Sequential DAC/DP treatment may be a novel strategy to simultaneously augment MAGE-3 expression and induce growth arrest in thoracic malignancies.




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