Anti-tumor Research Of Transfection With PcDNA3.1-HLA-A*0201 To Lewis Carcinoma Cells

【Objective】:HLA plays an important role in human immune system,espec ially in tumor occurrence and development.We chose the high frequent ge ne locus,HLA-A*0201 which is correlated to lung cancer.In this study,w e observe HLA-A2 negative-expressing Lewis lung cancer cells,which are t -ransfected with PcDNA3.1-HLA-A*0201 by using the liposomes method. The transfected cells were injected subcutaneouly into the C57BL/6 mice t o determine whether HLA-A*0201 restricted,CD8~+-mediated anti-tumor resp onse could be elicited in vivo.As a result,we were likely to provide the evidence for the tumor-immunotherapy mediated by HLA-restricted,CD8-me diated anti-tumor response.【Method】:Lewis lung cancer cells are cultured;gene-transfection mediated by liposomes is performed;the HLA-A2 positive-expressing cells are inje cted subcutaneouly into the mice;then the effect of anti-tumor effect was o bserved;flow cytometry(FCM)is used to detect the immune function in th e peripheral blood of each group.【Results】:We choosede HLA-A*0201 gene negative Lewis lung cancer cells.We transtected Lewis lung cancer cells with PcDNA3.1-HLA-A*0201 and acquired the positive cell clone hy G418,and injected them subcutaneouly into the C57BL/6 mice,then performed the cytotoxicity test in vivo.As a result,we showed that CD8-mediated cytotoxicity to Lewis lung cancer cells transfected with HLA-A~*0201 was significantly increased compared with nontransfected and GFP transfected groups.【Conclusion】:In vivo,transfed PcDNA3.1-HLA-A~*0201 into HLA-A~*0201 negative tumor cells could induce HLA-A~*0201 restricted,CD8-mediated anti-tumor response.