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Objective To investigate the cytotoxicity and radiosensitization of 5-nitroindazole-3-formyliminodiacetic acid on HeLa cells.Methods HeLa cells in exponential growth phase were incubated in culture media with different doses and the survival rate was determined by MTT assay.The survival rate of cells receiving radiation combined with different doses of medicine was compared with that of the control.Results The cytotoxicity of S-nitroindazole-3-formyliminodiacetic acid on HeLa cells was very low.The drug had hypoxia radiosensitizing effect on HeLa cells.At doses of 0,6,12,24,48 and 96 μg/ml under hypoxia ,the survival rate were 0.91 ,0.87,0.84,0.81 ,0.76 and 0.60,respectively.At the dosage of 48 and 96 μg/ml,the survival rate were 0.85 and 0.73 under oxygenous).Conclusions 5-Nitroindazole-3-formyliminodiacetic acid has low cytotoxicity and rediosensitizing effect on HeLa cells.
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Objective To screen and isolate the radioresistance related genes of IRM-2 mice.Methods cDNA library of IRM-2 mice was constructed by SMART technique.Total RNA was isolated from spleens of IRM-2 male mice.The first-strand cDNA was synthesized by using PowerScript reverse transeriptase,and double-strand cDNA was synthesized and amplified by long PCR.The PCR products were purified,digested with restriction enzyme Sfi I.The ds-cDNA fragment lessthan 500 bp was fractionated and ligated to the Sfi I-digested pDNR-LIB vector.The ligation mixture was transformed into E.coil DH5α by electroporution transformation to generate the unamplified cDNA library.The quality of cDNA library was identified by PCR technique.130 clones from cDNA library were sequenced and compared with GenBank database.Results The cDNA library contained 2.25 x 106 independent clones with an average insert size of 1.2 kb.The ratio of recombination and full-length was 95% and 55%,respectively.21 pieces of EST sequences from cDNA library were not the same as the known mice genes and registered into GenBank EST database,with registered number DW474856-DW474876.Conclusions cDNA library of IRM-2 mice has been constructed successfully.21 pieces of EST implies that radioresistance correlative genes may be in IRM-2 mice,which will lay a foundation for isolating and identifying radioresistance related genes in further study.