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目的探讨一种新齐墩果酸衍生物(3β-hydroxyolea-12-en-28-oic acid-3,5,6-trimethylpyrazin-2-methyl ester,简称为受试物)对人肝细胞癌细胞株SMMC-7721细胞的抗肿瘤作用机制。方法应用四甲基偶氮唑蓝(MTT)法及细胞形态学观察新齐墩果酸衍生物对SMMC-7721细胞增殖及形态的影响;Annexin V联合PI染色法检测新齐墩果酸衍生物对细胞膜以及用JC-1染色法检测线粒体膜电位的损伤程度;Western blotting检测新齐墩果酸衍生物对凋亡相关蛋白Bcl-2,Bax,caspase-9 and caspase-3的表达。结果新齐墩果酸衍生物能抑制人SMMC-7721细胞增殖力,与浓度和时间呈正相关,作用细胞48 h后,细胞出现核固缩,细胞裂解,产生凋亡小体等形态学改变,细胞膜完整性被破坏,线粒体膜电位(Δψm)被降低,Bax、pro-caspase-9、cleaved caspase-9、pro-caspase-3以及cleaved caspase-3蛋白表达水平被上调,Bcl-2蛋白表达水平被下调。结论这种新齐墩果酸衍生物在人SMMC-7721细胞中发挥抗癌作用可能与其破坏了线粒体功能有关。
Objective To investigate the effect of 3β-hydroxyolea-12-en-28-oic acid-3,5,6-trimethylpyrazin-2-methyl ester (abbreviated as test substance) on human hepatocellular carcinoma Antitumor mechanism of strain SMMC-7721 cells. Methods MTT assay and cell morphology were used to observe the effects of the new oleanolic acid derivatives on proliferation and morphology of SMMC-7721 cells. Annexin V and PI staining were used to detect the effects of the new oleanolic acid derivatives The damage of mitochondrial membrane potential was detected by JC-1 staining, and the expression of Bcl-2, Bax, caspase-9 and caspase-3 were detected by Western blotting. Results The new oleanolic acid derivatives could inhibit the proliferation of human SMMC-7721 cells in a dose-dependent manner with concentration and time. After 48 h, the cells exhibited nuclear pyknosis, cell lysis and morphological changes such as apoptotic bodies, The membrane integrity was destroyed, the mitochondrial membrane potential (Δψm) was decreased, the protein expression of Bax, pro-caspase-9, cleaved caspase-9, pro-caspase-3 and cleaved caspase- It is lowered. Conclusion The novel oleanolic acid derivatives play an anti-cancer role in human SMMC-7721 cells may be related to their destruction of mitochondrial function.