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Báo cáo khoa học: "Biological characteristics of Chinese hamster ovary cells transfected with bovine Prnp"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Biological characteristics of Chinese hamster ovary cells transfected with bovine Prnp | J. Vet. Set. 2007 8 2 131-137 JOURNAL OF Veterinary Science Biological characteristics of Chinese hamster ovary cells transfected with bovine Prnp Sang-Gyun Kang Deog-Yong Lee Mi Lan Kang Han Sang Yoo Department of Infectious Diseases KRF Zoonotic Disease Priority Research Institute and BK21 Program for Veterinary Science College of Veterinary Medicine Seoul National University Seoul 151-742 Korea A normal prion protein PrPc is converted to a proteaseresistant isoform by an apparent self-propagating activity in transmissible spongiform encephalopathy a neuro-degenerative disease. The cDNA encoding open reading frame ORF of the bovine prion protein gene Prnp was cloned from Korean cattle by PCR and was transfected into Chinese hamster ovary CHO-K1 cells using lipofectamine. The gene expression of the cloned cDNA was confirmed by RT-PCR and Western blotting with the monoclonal antibody 6114. Cellular changes in the transfected CHO-K1 cells were investigated using parameters such as MTT lactate dehydrogenase LDH and superoxide dismutase SOD activities as well as nitric oxide NO production and an apoptosis assay. In the MTT and LDH assays the bovine Pr iP-transfectant showed a lower proliferation rate than the wild-type p 0.05 . Production of NO after LPS or ConA stimulation was not detected in either transfectants or CHO-K1 cells. In SOD assay under ConA stimulation the SOD activity of transfectants was 10 times higher than that of CHO-K1 cells at 6 h after treatment Ip 0.05 . The genomic DNA of both the transfectants and control cells began to be fragmented at 611 after treatment with cyclohexamide. Caspase-3 activity was reduced by transfection with the bovine Prop p 0.05 . Conclusively the viability of transfectants expressing exogenous bovine Prop was decreased while the capacities for cellular protection against antioxidative stress and apoptosis were increased. Key words BSE CHO-K1 cells Korean cattle prion Prnp Introduction Bovine spongiform encephalopathy BSE