熊果酸诱导人肝癌SMMC-7721细胞株凋亡机制的研究

Mechanism of ursolic acid induced apoptosis in human hepatoma cell line SMMC-7721

目的:研究熊果酸(ursolic acid,UA)对人肝癌SMMC-7721细胞增殖抑制作用及诱导凋亡的相关机制。方法:MTT法测定UA对SMMC-7721细胞的生长抑制作用;Wright-Giemasa染色和Hoechst33258荧光染色法观察细胞的形态学变化;流式细胞仪检测细胞的凋亡率;RT-PCR检测细胞凋亡相关基因p53和Fas的表达。结果:UA对SMMC-7721细胞生长有明显的生长抑制作用,呈明显的时间-效应关系和浓度-效应关系,24 h的半数抑制浓度(IC50)为45.72μmol/L。Wright-Giemasa染色、Hoechst荧光染色证实了UA可以通过凋亡途径诱导SMMC-7721细胞发生死亡。流式细胞仪结果进一步证实了UA可以诱导SMMC-7721细胞凋亡,而且凋亡率增加呈一定的量效和时效关系。RT-PCR结果显示UA能够上调p53和Fas基因的表达。结论:UA在体外对SMMC-7721细胞具有抗增殖和诱导凋亡的作用。其机制可能与p53和Fas基因的变化有关。

AIM： To study the inhibitory effect and apoptosis mechanism of ursolic acid（UA） on human hepa-toma cell line SMMC-7721.METHODS： The growth inhibition of SMMC-7721 cells was assessed by MTT colori-metric assay.Morphological changes of cells were evaluated Wright-Giemasa staining and Hoechst 33258 staining after it was induced by UA.Flow cytometry was used to detect the apoptotic rate.The expression of aposptosis re-lated to genes p53 and Fas was evaluated by RT-PCR.RESULTS： UA strongly inhibited the proliferation of SMMC-7721 cells in a dose-and time-dependent manner.The IC50 in 24 h was 45.72 μmol/L.UA-induced apop-tosis was confirmed by Wright-Giemasa staining and Hoechst 33258 staining.Flow cytometric analyses showed UA could induce SMMC-7721 cells apoptosis and the rate of apoptosis could be increased in a time-and dose-depend-ent manner.The genes expression of p53 and Fas increased in SMMC-772 cells cultured with UA.CONCLU-SION： UA has the inhibitive effect on SMMC-7721 cells proliferation and induction of cell apoptosis related with the expressions of p53 and Fas.