Antitumor Research Of Transfection With PcDNA3.1-HLA-A2 To Yunnan Gejiu Lung Adenocarcinoma Cells (GLC-82)

Objective: HLA plays an important role in human immune system, especially in tumor occurrence and development. Data about HLA class I alleles and related diseases were limited. To remedy the lack of these data, we analysed HLA gene polymorphism of Yunnan lung cancer cells and chose the high frequent gene locus, HLA-A2 correlated to lung cancer. In this study, we observed Yunnan Gejiu lung adenocarcinoma cells(GLC-82) transfected with PcDNA3.1-HLA-A2 using liposomes to determine whether HLA-A2 resticted, CD8-mediated antitumor response could be elicited in vitro. Method: Screening of HLA-A2 gene negative expressing lung cancer cells by cell culture, inverted fluorescence microscope, Polymerase Chain Reaction-Sequence Specific Primers(PCR-SSP), etc. Lung cancer cells transfected with PcDNA3.1-HLA-A2 using catholyte liposomes vector and aquired stable positive transfected cell clone by G418. Flow cytometry analysed the transfected efficiency and observed positive transfected cells' morphology, cycle change. Collected these cells and CD8+ T cells separated by immunomagnetic beads to carry out cytotoxicity test. Lactate dehydrogenase(LDH) assay was employed to eveluate the cytotoxic effect. Results: Acquired HLA-A2 gene negative expressing lung cancer cells,GLC-82 and HLA-A2 gene positive lymphocytes from healthy donor. Transfected GLC-82 with PcDNA3.1-HLA-A2 and screened out the positive cellclone by G418 , Flow cytometric analysis of positive cell clone showed a significant change from common GLC-82. Cytotoxicity test in vitro showed that CD8-mediated cytotoxic effect to GLC-82 transfected with PcDNA3.1-HLA-A2 signifiantly increased. Conclusion: HLA-A2 gene deletion or no expressing is an important reason for tumor cells' escaping from human immune surveillance. In vitro, transfection with PcDNA3.1-HLA-A2 to HLA-A2 negative expressing tumor cells can induce HLA-A2 restricted, CD8-mediated antitumor response.