鸦胆子油乳诱导肝癌细胞凋亡及对相关基因表达的影响  

Effects of seminal oil emulsion of Brucea javanica on apoptosis and apoptosis- related genes in human hepatocellular carcinoma cells

作　　者：马力[1] 张月宁[1] 

机构地区：[1]兰州医学院第二附属医院消化内科,甘肃省兰州市730030

出　　处：《世界华人消化杂志》2004年第3期559-562,共4页World Chinese Journal of Digestology

摘　　要：目的:研究鸦胆子油乳体外诱导人肝癌细胞SMMC-7721 凋亡的作用及对细胞周期和凋亡相关基因p53和Bcl-2表达的影响,探讨其抗肿瘤机制. 方法:MTI怯检测鸦胆子油乳不同浓度和作用时间对肝癌细胞的抑制增生作用;透射电镜观察细胞形态学改变;琼脂糖凝胶电泳分析DNA特征;流式细胞仪检测细胞凋亡和细胞周期分布;免疫细胞化学染色检测p53和Bcl-2的表达. 结果:鸦胆子油乳对人肝癌细胞SMMC-7721具有显著的抑制增生作用,且有时间和浓度依赖性.透射电镜和凝胶电泳可观察到凋亡特征性的形态学和生化特征改变.0.10 g/L 鸦胆子油乳作用12,24,48h后,流式细胞仪分析可见典型的亚二倍体峰,细胞周期阻滞于G0/G1期,与对照组相比有显著性差异(P<0.05).p53和Bcl-2在经鸦胆子油乳作用后表达水平均下降,二者呈正相关(r=0.966,P<0.05), p53下降更为明显. 结论:鸦胆子油乳体外对肝癌细胞SMMC-7721有显著的抑制增生作用,能诱导凋亡、阻滞细胞周期于G0/G1期,抑制p53和Bcl-2的表达是其重要机制,其中p53途径起主导作用.AIM: To explore the anti-tumor mechanism of seminal oil emulsion of Brucea javanica by studying in vitro its effects on apoptosis, cell cycle and expression of apoptosis-related genes p53 and Bcl-2 in human hepatocellular carcinoma cell line SMMC-7721. METHODS: Anti-proliferation effect was measured by MTT assay. The morphology of cells was observed under transmission electron microscope. Agarose gel electrophoresis was used to analyze DNA character and the flow cytometry to detect apoptotic rate and cell cycle distribution. The levels of p53 and Bcl-2 protein were examined by immu-nocytochemical staining. RESULTS: The proliferation of hepatocellular carcinoma cell line SMMC-7721 could be remarkably inhibited by seminal oil emulsion of Brucea javanica in a time- and concentration-dependent manner. Morphological and biochemical changes characteristic of apoptosis were observed through electron microscope and agarose gel electrophoresis. After 12, 24, 48 h incubation with 0.10 g/L emulsion of seminal oil of Brucea javanica, the flow cytometry showed typical subdiploid peaks and the cell cycle was arrested at G0/G1 phase(P<0.05). The expression of p53 and Bcl-2 was down-regulated after exposure to the drug, with a positive correlation between them (r =0.966, P<0.05). CONCLUSION: Seminal oil emulsion of Brucea javanica can significantly inhibit the proliferation of human hepatocellular carcinoma cell line SMMC-7721 in vitro through inducing apoptosis and arresting cell cycle at G0/G1 phase, and its underlying mechanism is related to the down-regulating mutant type p53 as well as Bcl-2, with p53 pathway playing a leading role.

关 键 词：鸦胆子油乳 肝癌 细胞凋亡 凋亡相关基因 MTT法 流式细胞仪 p53 Bcl-2 

分 类 号：R735.7[医药卫生—肿瘤]