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目的:探讨DFO、5-ALA不同浓度和pH值对体外诱导大肠癌细胞原卟啉Ⅸ聚积的影响,为该药物诱导大肠癌组织原卟啉Ⅸ聚积用于大肠早癌的诊断提供依据。方法:pH值分别为5.5、6.0、6.5、7.0、7.5、8;5-ALA浓度分别为0.0、0.5、1.0、1.5、2.0、2.5、3.0moml/mL;DFO浓度分别为0.0、5、10、15、20、30moml/mL的诱导培养液分别培养2小时后终止培养。高效液相色谱-荧光检测仪测定细胞内原卟啉Ⅸ含量。结果:pH值为7.0时原卟啉Ⅸ含量最高,与pH﹤6.5或pH≥7.5各组相比有显著差异(P﹤0.05);DFO浓度为10moml/mL时,5-ALA的最适诱导浓度为1.5～2.0moml/mL;5-ALA浓度为2moml/mL时,DFO的最适诱导浓度为10～20moml/mL。结论:5-ALA的最适浓度为1.5～2.0moml/mL,DFO的最适浓度为10～20moml/mL;诱导药物缓冲液的最适pH值为6.5～7。
OBJECTIVE: To investigate the effect of different concentrations of DFO and 5-ALA and pH value on the accumulation of protoporphyrin Ⅸ in colorectal cancer cells in vitro, and to provide a basis for the drug-induced accumulation of protoporphyrin Ⅸ in colorectal carcinoma for the diagnosis of early colon cancer. Methods: The pH values were 5.5,6.0,6.5,7.0,7.5,8 respectively; the concentrations of 5-ALA were 0.0,0.5,1.0,1.5,2.0,2.5,3.0moml / mL; the concentrations of DFO were 0.0,5,10 , 15,20,30 moml / mL of the culture medium were cultured for 2 hours after termination of culture. Determination of Protoporphyrin IX in Cells by High Performance Liquid Chromatography with Fluorescence Detection. Results: The content of protoporphyrin IX was the highest at pH 7.0, which was significantly different from that at pH 6.5 or pH 7.5 (P <0.05). When DFO was 10moml / mL, the optimal induction of 5-ALA Concentration of 1.5 ~ 2.0moml / mL; 5-ALA concentration of 2moml / mL, DFO optimum induction concentration of 10 ~ 20moml / mL. CONCLUSION: The optimal concentration of 5-ALA is 1.5-2.0moml / mL and the optimum concentration of DFO is 10-20moml / mL. The optimum pH value of the induced drug buffer is 6.5-7.