【摘要】 目的观察柴胡皂甙d(SSd)对肝癌SMMC-7721细胞环氧合酶-2(COX-2)蛋白表达和细胞内Ca2+变化的影响,探讨其抗癌作用机制。方法肝癌SMMC-7721细胞株经不同终浓度的SSd(2.5、5.0、10.0和20.0 mg/L)作用48 h后,MTT比色法检测细胞增殖抑制率,荧光染色法观察其凋亡状况,Western blotting检测COX-2表达,放射免疫法检测前列腺素E2含量,免疫荧光分光光度计法检测细胞内钙离子浓度。结果 SSd对肝癌SMMC-7721细胞生长、增殖有明显抑制作用,并诱导其凋亡。同时,SSd可以降低肝癌细胞COX-2蛋白表达和前列腺素E2含量水平,提高胞内钙离子浓度。在2.5～10.0 mg/L作用终浓度范围内,上述作用呈现一定的量效关系。结论提高细胞内钙离子浓度,可能是SSd抑制COX-2活性,诱导人肝癌细胞凋亡的机制之一。更多还原

【Abstract】 【Objectives】 To observe the effect of Saikosaponins-d(SSd) on COX-2 expression and intracellular calcium in human hepatoma carcinoma cell line SMMC-7721 and to investigate the mechanism of antitumous effect.【Methods】 The human hepatoma carcinoma cell line SMMC-7721 was cultured in vitro and then treated with SSd of various concentrations(2.5,5.0,10.0,20.0 mg/L) for 48 hrs,and the inhibitory rate of cell proliferation was measured by MTT.The cell apoptosis was observed by staining with Hoechst 33342,and the expression of Cyclooxygenase-2(COX-2) protein in the cells was determined by Western bolt.Prostaglandin E2(PGE2) in medium was detected by radioimmunoassay,and the concentration of intracellular calcium was tested with spectrofluorometer.【Results】 SSd could obviously inhibit the growth and proliferation of SMMC-7721 cells,and induced their apoptosis.Meanwhile,SSd could depress the expression of hepatoma carcinoma cell COX-2 protein,cut down PGE2 level and increase intracellular calcium concentration.There was a dose-effect relationship when the SSd concentration was in the range of 2.5 to 10.0 mg/L.【Conclusion】 Improve intracellular calcium concentration might be one of the mechanisms of inhibiting COX-2 activity and inducing SMMC-7721 apoptosis by SSd.更多还原