5-aza-2'-deoxycytidine (DAC) treatment induces the MAGE-A10 expression and improves the cytotoxicity of MAGE-A10-specific CTLs in lung cancer cells.

Abstract

BackgroundMAGE-A10 is a subtype of the Melanoma-associated antigen A (MAGE-A), a class of tumor antigens that are extensively expressed in various histological types of tumors and represents an attractive target for tumor immunotherapy. Epigenetic-modifying drugs can enhance the expression of tumor antigens and improve the cytotoxicity of antigen-specific T cells. 5-aza-2’-deoxycytidine (DAC), a DNA methyltransferase inhibitor (DNMTI) considered an epigenetic-modifying drug, could enhance the expression of MAGE-A10 in cancer cells.MethodsHuman lung cancer cell lines (H1975 and A549) and primary lung cancer cells (L228, L329 and L419) were used. 5-aza-2’-deoxycytidine was used to induce the expression of MAGE-A10 in tumor cells. MAGE-A10 antigenic peptide (sequence: SLLKFLAKV) was used to induce differentiation of MAGE-A10-specific cytotoxic T lymphocytes (CTLs). Interferon-γ release assay was used to detect the capacity of MAGE-A10 peptide to induce CTLs. Cell Counting Kit-8 (CCK-8) analysis was performed to detect the cytotoxicity of MAGE-A10-specific CTLs. Real-time PCR and western blot analysis was used to detect the mRNA and protein levels, respectively. Immunohistochemistry was performed to detect the protein expression in cancer and adjacent normal tissue. Kaplan-Meier plotter online database was used to analyze the overall survival (OS), post-progression survival (PPS), and first progression (FP).ResultsThe lysis rate of MAGE-A10-specific CTLs in L419 and H1975 were found to be 65.9% and 80.5%, respectively. Both L419 and H1975 showed significantly higher lysis rate in group 1 than in group 3 (6.7, 26.7%), group 2 (0, 0%) and group 4 (0, 0%) (P=0.0003, P≤0.0001, P≤0.0001, respectively). Online data mining using Kaplan-Meier plotter suggested that high expression of MAGE-A10 was significantly and negatively associated with OS (Plogrank =2.1e-05) and PPS (Plogrank =0.0057), and FP (Plogrank =3.2e-12).ConclusionsHigh-level expression of MAGE-A10 improved the anti-tumor immune cytotoxicity of MAGE-A10-specific CTLs in lung cancer cell lines and primary lung cancer cells. However, MAGE-A10 gene expression was negatively associated with prognosis according to the survival analysis. Thus, we hypothesize that high-level of MAGE-A10 expression in vivo may inhibit the differentiation of MAGE-A10-specific CTLs.