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Archive > Volume 30 Issue 8 > 2023,30(8):701-706. DOI:10.3872/j.issn.1007-385X.2023.08.008 Prev Next
Clinical Research
Killing effects of 5-Aza-CdR combined with EBNA1-DC vaccine-induced lymphocytes on nasopharyngeal carcinoma C666-1 cells
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Abstract:
Objective: To investigate the killing effects of Epstein-Barr virus nuclear antigen 1 (EBNA1) mRNA-modified DC (EBNA1-DC) -induced lymphocytes combined with methylation inhibitor 5-Aza-CdR on nasopharyngeal carcinoma C666-1 cells.Methods: EBNA1-pCDNA3.1 plasmid was used as a template, and EBNA1 mRNA was obtained by in vitro transcription.Subsequently, EBNA1 mRNA was transfected into dendritic cells derived from peripheral blood from healthy donors by liposome toconstruct EBNA1-DC vaccine. Flow cytometry was used to detect the transfected DC phenotype and the apoptosis of C666-1 cells after 5-Aza-CdR treatment. Real-time cell analysis was used to detect the specific antitumor activity of EBNA1-DC vaccine-induced lymphocytes combined with 5-Aza-CdR. Results: The positive rate of EBNA1on the surface of EBNA1-DC after transfection with EBNA1 mRNA was (59.3±5.85) %. The expression of HLA-DR was significantly higher than that of untransfected DC ([84.9±5.5]% vs [68.0±5.8]% , P=0.026). The expression of CD80 was significantly improved from (88.2±3.9)% to (61.1±4.4)% (P=0.015). The apoptosis of C666-1 cells treated with low-dose 5-Aza-CdR was not significantly different from that of untreated cells. The expression of IRF7 gene in C666-1 cells pretreated with low-dos 5-Aza-CdR was significantly higher than that in untreated cells (P=0.000 1).Lymphocytes induced by EBNA1-DC had stronger specific killing activity against EBV+C666-1 cells compared with untransfected DC (P=0.049). C666-1 cells pretreated with low dose 5-Aza-CdR were more sensitive to specific immune killing induced by EBNA1-DC (P=0.019). Conclusion: The combination of 5-Aza-CdR and EBNA1-DC vaccine can significantly enhance the specific immune killing effect on C666-1 cells. This study provides the preliminary research basis for the development of the mRNA-DC vaccine and its application in clinical comprehensive therapy.
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