The Immological Rejection Activated by Human Heptocarcinoma Transferred with Murine H-2K~b Gene / 中国肿瘤生物治疗杂志

ABSTRACT

Objective: To strengthen the immunogenicity of hepatocarcinoma cells and activate immnological cells recognizing and killing the tumor cells.Methods:The murine MHC-I gene H-2Kb which can express immunologial rejection antigens was transfected into human hepatocarcinoma cells HepG2 by liposome DNA mediated gene gene transfer.The transfection of H-2Kb gene were detected by molecular hybridization techniues.The exogeous antigens expressed on the membrane of trans- fected tumor cells were detected with ABC immunohistochemical method and flow cytometer. [3H] release assays were used to detect the recognizing and killing effects of lymphocytes to HepG2 cells transferred with murine H-2Kb gene. The nude mice ex- periment was used to further verify CTL cells killing active.Results:Southern blot hybridization showed that the H-2Kb gene was integrated into the chromosome of HepG2 cells. The RNA dot blot hybridization showed that there was transcription of H-2Kb DNA in the transfected tumor cells.ABC immunohistochemical method and flow cytometer detection showed that the murine H-2Kb antigens were expressed on the membrane of HepG2 cells. [3H] release assays showed that the cytotoalcyty to HepG2 cells fected with H-2Kb gene was obviously higher than that to control cells.The results demonstrated that the growth of hepatocarcino- ma cells which were transferred with H-2Kb gene was obviously inhibited.Conclusion:The murine MHC-I gene H-2Kb could be transferred into the human hepatocinoma cells and expressed on the membrane of transferred cells.The HepG2 cells transferred with H-2Kb gene could induce human effective lymphocytes to recognize and kill these transferred tumor cells.