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《Chinese Journal of Difficult and Complicated Cases》 2017-04
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Dual effects of nitric oxide on radiation induced cellular apoptosis and DNA double strand breaks in tumor cells

SU Xiaoming;CUI Di;REN Ye;CHEN Fang;MA Huizhen;DAI Zhuojie;WANG Zongye;Department of Radiation Oncology,306th Hospital of PLA;  
Objective To investigate the effects of different concentrations of nitric oxide(NO) on apoptosis and DNA double strand breaks in tumor cells with different p53 gene status.Methods The experiment was carried out in the laboratory of radiation medicine,The Fourth Military Medical University from June 2009 to June 2010.The p53 of human nonsmall cell lung cancer H1299 cells,stably transfected with wild-type or mutant p53(wtp53) p53(mp53) gene,H1299/wtp53 or H1299/mp53 cells.Analysis by RT-PCR and restriction fragment length polymorphism expression vectors in the cells was stable(RFLP) detection method.The cells were cultured in medium containing different concentrations of isosorbide dinitrate(ISDN),and then received X-ray irradiation.After irradiation,apoptosis and gamma H2AX were measured by Hoechst33342 staining and flow cytometry,respectively.The expression of apoptosis related protein was detected by Western blot.Results In H1299/wtp53 and H1299/mp53 cells,observed a clear band of 110 bp RT products can be PCR in exon 7of the p53 gene.Exon 7 of p53 gene with MspI or BsrI restriction endonuclease digestion specificity,corresponding in H1299/wtp53 or H1299/mp53 cells,can clearly observe the band of 70 bp.In H1299/wtp53 cells,the low concentration of ISDN(5 μmol/L) can significantly reduce the X-ray radiation induced apoptosis and DNA double strand breaks,whereas high concentrations of ISDN(500 μmol/L) was significantly increased apoptosis and DNA X-ray radiation induced double strand breaks.Furthermore,nitric oxide has bidirectional effects on radiation-induced Caspase 3 and Bax mediated apoptosis.However,for these concentrations of ISDN,none of these phenomena were observed in H1299/mp53 cells.Conclusion In tumor cells,changes of nitric oxide at different concentrations can be dual effects of radiation induced Bax and Caspase 3 mediated apoptosis and DNA double strand breaks,and nitric oxide through the regulation of p53 gene expression to affect cell fate.
【Fund】: 国家自然科学基金资助项目(81402577)
【CateGory Index】: R730.5
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