去铁胺联合5-氨基酮戊酸诱导人结肠腺癌SW480细胞原卟啉Ⅸ积聚的实验研究

Researches on cumulation of protoporphyrin Ⅸ in SW480 cells induced by DFO and 5-ALA

目的探讨去铁胺联合5-氨基酮戊酸体外诱导大肠癌细胞原卟啉Ⅸ积聚的特性。方法SW480细胞用6孔培养板分A、B、C、D4组分别用:含DFO0.1mmol/mL、含0.1mmol/mLDFO+2mmol/mL5-ALA、含5-ALA2mmol/mL、不含DFO、5-ALA的RPMI1640培养液常规培养10h,荧光显微镜观察SW480细胞内原卟啉Ⅸ的荧光分布及强度,高效液像色谱-荧光检测器定量分析SW480细胞内原卟啉Ⅸ的含量。结果B组、C组细胞质内可见不同强度的红色荧光,B组细胞内原卟啉Ⅸ的荧光强度明显强于C组,高强度荧光细胞计数分别占(57.67±2.49)%和(20.07±2.05)%,差异有显著性(P<0.001);4组之间SW480细胞内原卟啉Ⅸ的含量有明显差异:A组<D组<C组<B组。结论去铁胺联合5-氨基酮戊酸体外诱导可有效增加大肠癌细胞内原卟啉IX的积聚。

[Objective] To explore the characteristics of cumulation of protoporphyrin Ⅸ in SW480 cells induced by DFO and 5-ALA. [Method] SW480 cells were respectively cultivated in 4 groups, Group A, B, C, D, using culture solution, RPMI1640 with DFO 0.1 mmol/mL, 0.1 mmol/mL DFO ＋ 2 mmol/mL 5-ALA, 5-ALA 2 mmol/mL, without DFO, 5-ALA for 10 hr. Fluorescence distribution and intensity of protoporphyrin Ⅸ in SW480 cells were observed under fluorescence microscope, the content of protoporphyrin Ⅸ in SW480 cells were quantitatively analyzed by high performance liquid chromatography. [Results] Different intensities of red fluorescence can be observed in SW480 from Group B and Group C. The fluorescence intensity of protoporphyrin Ⅸ from Group B is significantly higher than that from Group C. The count of high intensive fluorescence cells axe respectively （57.67± 2.49）% and （20.07±2.05）%, with significant difference （P 〈0.001）; the content of protoporphyrin Ⅸ in SW480 cells from 4 groups are different, Group A〈 Group D〈 Group C〈 Group B. [Conclusion] It could efficiently increase cumulation of protoporphyrin Ⅸ in SW480 cells induced by DFO and 5-ALA.