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《Chinese Journal of Cancer》 2001-10
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Initiated Cancerazation Effect of Reactive Oxygen Species and Its Produ cts of Oxidative DNA Damage in B EP2D Cells

ZHU Mao-xiang * ,YANG Zhi-hua,GONG Yi-fen,LU Ying,CAO Zhen-shan Institute of Radiatio n Medicine,Academy of Military Medical Science,B eijing 100850,P.R.Ch ina  
Objective:To investigate oxidative DNA damage through reactive o xygen species(ROS)produced by 60 Co gamma ray irradiation or 4-(methylnitrosamino)-1-(3-pyridyl) -1-butanone(NNK)treatment in HPV-16immortalized hu man bronchial epithel ial cell(BEP2D),and their effect in initiating cancerazation.Methods :Ethi dium bromide and 2',7'-dichlorofluorescein,fluorescent products of the mem brane-permeable dyes hydroethine and 2' ,7' -dichloroflurescin diacetate,w ere used to monitor the i ntracellular production of hydrogen peroxides(H 2 O 2 )and superoxide anions(O 2 )respectively,by flow cytometry.8-hydroxydeoxygunosine(OH8dG)was e xamined with HPLC-ECD from extr acted DNA.DNA from BEP2D irradiated by 60 Co gamma ray or treated with NNK were t ransferred into receptor C3H10T1 /2cells together with coli-neo plasmid thro ugh liposome,and positive clone of t ransfection was selected with Geneticin(G418).Cell transformation fre quency was a ssayed by counting clone formation i n semisolid agar,and carcinog enesis was evaluated by tumor formation in nude mice.Results:The ROS products and DNA adduct OH8dG in 60 Co gamma ray irradiated or NNK-treated BEP2D cells increased remarkably,a nd showed better dose-response correlation.After 60 Co gamma rays irradiated or NNK-treate d DNA transfection,the receptor C 3H10T1/2cells displayed the characte ristics of transformation,and tumor formatio n was observed in nude mice.Conclusion:Intracellular production of RO S and oxidative DNA damage induced by radi ation and /or chemical carcinogen ma y initiate ce ll cancerazation.
【Fund】: 国家自然科学基金资助项目(批准号:39700030)
【CateGory Index】: R730.2
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