摘要
目的:探讨全反式维甲酸、18β-甘草次酸和甘草酸对BEL-7402人原发性肝癌细胞诱导分化和凋亡的作用。方法:用此三种药物处理BEL-7402人肝癌细胞,观察细胞增殖,测定核质比例、鸟氨酸氨基甲酰转移酶(OCT)、酪氨酸-α-酮戊二酸转氨酶(TAT)、碱性磷酸酶(ALP)和γ-谷氨酰转肽酶(γ-GT)活性,用RIA法测定甲胎蛋白(AFP)分泌量,用吖啶橙(AO)和溴乙锭(EB)荧光双染法和末端脱氧核苷酸转移酶介导dUTP缺口末端标记法(TUNEL法)检测凋亡细胞。结果:维甲酸、18β-甘草次酸和甘草酸对肝癌细胞增殖均有抑制作用,并有量效关系,其IC_(50)分别为16.29μmol/L、78.23μmol/L和3.94mmol/L。10μmol/L维甲酸、60μmol/L 18β-甘草次酸和2.5mmol/L甘草酸均使肝癌细胞核质比例显著下降(P<0.05和P<0.01),并使代表肝癌细胞分化的酶OCT、TAT和ALP比活力明显升高,使AFP分泌量和γ-GT比活力明显下降。60μmol/L、90μmol/L 18β-甘草次酸、20μmol/L维甲酸和5mmol/L甘草酸均使肝癌细胞凋亡率明显升高(P<0.05和P<0.01)。结论:维甲酸、18β-甘草次酸和甘草酸有抑制肝癌细胞增殖和诱导其分化的作用,且在同等效果下18β-甘草次酸所需的浓度比甘草酸的浓度约低40倍;三种药物均有诱导肝癌细胞凋亡的作用。
Objective: To investigate the effects of all-trans retinoic acid, 18β-glycyrrhetinic acid and glycyrrhizin on the induction of differentiation and apoptosis of BEL-7402 human primary hepatocarcinoma cells. Methods: The BEL-7402 cells were treated with three kinds of the drugs to observe the variations of cell proliferation and to examine the nucleocytoplasm ratio and the specific activities of omithine carbamoyl transferase (OCT), tyrosine-α-ketoglutarate transaminase (TAT), alkaline phosphatase (ALP), and γ-glutamyl transpeptidase (γ-GT) of the cells. The alpha-fetoprotein (AFP) secretary amount of the cells were assayed by radioimmunoassay (RIA). The apoptosis of the cells was detected by the method of acridine orange-ethid-ium bromied fluorescent stain and TUNEL. Results: After treated with retinoic acid, 18β-glycvrrhetinic acid and glycyrrhizin, the proliferations of hepatocarcinoma cells were all inhibited, and the inhibition degrees were related to dose of the drugs. IC50 of the proliferation inhibition of three kinds of the drugs was 16.29 μmol/L, 78.24 μmol/L and 3.94mmol/L respectively. Treated with 10μmol/L retinoic acid, 60 μmol/L 18 β-glycyrrhetinic acid and 2.5mmol/L glycyrrhizin, the nucleocytoplasm ratios of the cells were all reduced significantly (P<0.05 and P<0.01), the specific activities of OCT, TAT and ALP that reflect the differentiation of liver cancer cell were all increased evidently, and the secretory amount of AFP and the specific activities of γ-GT were decreased obviously. The percent of apoptotic cells induced by 60μmol/L and 90μmol/L 18β-glycyrrhetinic acid, 20μ-mol/L retinoic acid and 5mmol/L glycyrrhizin were increased significantly (P<0.05 and P<0.01). Conclusion: Retinoic acid, 18β-glycyrrhetinic acid and glycyrrhizin can inhibit proliferation of BEL-7402 human hepatocarcinoma cells and induce differentiation of the cells. The dosage of 18β-glycyrrhetinic acid is lower about 40 times in the same effects than that of glycyrrhizin. Three kinds of the drugs can also induce apoptosis of the cells.
出处
《中西医结合肝病杂志》
CAS
2003年第3期148-150,共3页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
基金
广东省中医药管理局项目(GD-95-0612)
关键词
全反式维甲酸
18Β-甘草次酸
甘草酸
细胞分化
细胞凋亡
人原发性肝癌
Medical Oncology
Liver Neoplasms, Experimental
Drug Screening Assays, Antitumor
All-trans Retinoic Acid/Pharmacodynainics
18β-glycyrrhetinic Acid/Pharmacodynamics
Glycyrrhizin/Pharmacodynamics
Tumor Cells, Cultured
