喜树碱及其衍生物对肝癌细胞HepG2增殖抑制与凋亡的研究

摘要

Objective To explore the effect of camptothecin ( CPT) and its derivatives 10-hydroxy camptothe-cin (HCPT),7-ethyl camptothecin (SN22),7-ethyl-10-hydroxy camptothecin (SN38) on inhibition of hepatoma cell line HepG2 and induction of its apoptosis. Methods The vegetative HepG2 cells of logarithmic phase were assigned as control group and experimental group. Four drugs were made to different concentrations,which were 0. 01,0. 1,1,10, 100 μmol/L. Different concentrations of drug solutions were effected on HepG2 cells in 48 h,then MTT method was used to detect the proliferation of HepG2 cell and cell apoptosis was analyzed by flow cytometry ( FCM) using Anexin V-PI staining. Results The results by MTT method showed four drugs had inhibitory effect on proliferation of HepG2 cells,moreover,the proliferation inhibition of HepG2 cells was enhanced gradually with the increase of drug concentra-tions in a certain scope drug concentrations,and the proliferation inhibition was dependent on drug concentration. The sequences of proliferation inhibition were SN38,HCPT,SN22,and CPT. With different concentrations of camptothecin and derivatives with HepG2 liver cancer cells,there were significant differences compared with control group,and as drug concentration changed,cell apoptosis also presents gradient change;structure modification of SN38 was stronger than SN22,CPT and HCPT function by FCM using AnexinV-PI staining. Conclusion Camptothecin and its derivatives can inhibit HepG2 cells and induce cell apoptosis.%目的：探讨喜树碱(CPT)及其衍生物10-羟基喜树碱(HCPT)、7-乙基喜树碱(SN22)、7-乙基-10-羟基喜树碱(SN38)在体外对肝癌细胞HepG2增殖抑制与凋亡的影响。方法取对数期生长的HepG2细胞,分为对照组以及实验组,将4种药物配成浓度梯度0．01、0．1、1、10、100μmol/L的药物溶液。将不同浓度的药物溶液作用于肝癌细胞48 h,MTT法测定HepG2细胞的增殖情况,Anexin V-PI染色,流式细胞仪检测细胞凋亡情况。结果 MTT法显示,4种药物对肝癌细胞 HepG2的增殖具有抑制作用,在一定范围内随着药物浓度的增加,对HepG2细胞增殖的抑制作用逐渐增强,呈量效依赖关系,其中7-乙基-10-羟基喜树碱抑制效果最好,10-羟基喜树碱、7-乙基喜树碱次之,喜树碱抑制效果最差。不同浓度的喜树碱及衍生物处理肝癌细胞HepG2,与对照组比较差异有统计学意义,随着药物浓度的变化,细胞凋亡也呈现梯度变化,结构修饰后的SN38比SN22、HCPT及CPT作用更强。结论喜树碱及衍生物对肝癌细胞HepG2有抑制作用,并且能够诱导细胞凋亡。