Antitumor Research Of Transfection With PcDNA3.1-HLA-A～*0201 To Yunnan Gejiu Lung Squamous Carcinoma Cells(YTMLC)

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-A*0201 correlated to lung cancer. In this study, we observed Yunnan Gejiu lung squamous carcinoma cells(YTMLC) transfected with PcDNA3.1-HLA-A*0201 using liposomes to determine whether HLA-A*0201 restricted, CD8-mediated antitumor response could be elicited in vitro. Method: Screening of HLA-A*0201 gene negative expressing lung cancer cells and HLA-A*0201 gene positive expressing health donor by cell culture, invited fluorescence microscope. Polymerase Chain Reaction-Sequence Specific Primers(PCR-SSP), etc. Lung cancer cells transfected with PcDNA3.1-HLA-A*0201 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. LAK cells were induced culturing. Collected these cells and CD8~+Tcells separated by immunomagnetic beads to carry out cytotoxicity test. We observed these results by MTT method, flow cytometer. Results: Acquired HLA-A*0201gene negative expressing lung cancer cells, YTMLC and HLA-A*0201 gene positive activated lymphocytes from healthy donor. Transtected YTMLCwith PcDNA3.1-HLA-A*0201 and screened out the positive cell clone by G418, Flow cytometric analysis of positive cell clone showed a significant...