摘要
背景:薏苡仁提取液目前已广泛应用于临床抗肿瘤治疗,但至今尚未见其单独应用于胰腺癌的报道。目的:观察薏苡仁提取液对人胰腺癌细胞凋亡和超微结构的影响,以探索胰腺癌治疗的新途径。方法:以不同浓度薏苡仁提取液处理人胰腺癌细胞系PaTu-8988后,分别通过光镜观察、Hoechst33258荧光染色、末端脱氧核苷酸转移酶原位标记(TUNEL法)和流式细胞术检测细胞凋亡,应用透射电镜观察细胞超微结构的变化。结果:经10μl/ml和20μl/ml薏苡仁提取液作用72h后,PaTu-8988细胞的增殖能力明显下降,其作用呈剂量依赖性;光镜观察显示凋亡细胞变圆、体积缩小,Hoechst33258荧光强染,TUNEL标记阳性。20μl/ml薏苡仁提取液作用24、48和72h后,流式细胞仪DNA直方图上出现亚二倍体凋亡峰,细胞凋亡率呈时间依赖性,分别为7.1%±0.6%、11.3%±0.3%和15.1%±2.9%,显著高于对照组的2.4%±1.1%(P<0.01)。透射电镜观察显示,凋亡早期主要是线粒体结构的改变,后期则出现典型的凋亡征象,如核固缩、染色质凝集靠近核膜和凋亡小体形成。结论:薏苡仁提取液可诱导人胰腺癌细胞凋亡,其作用呈剂量和时间依赖性,线粒体可能在早期细胞凋亡中起重要作用。
Background: Coix seed extract is now widely used in the treatment of cancers clinically, however, its effect on pancreatic cancer has not been investigated yet. Aims: To observe the effects of coix seed extract on apoptosis and ultrastructure of human pancreatic cancer cells, so as to seek a new approach in treating pancreatic cancer. Methods: Human pancreatic cancer cell line PaTu-8988 was exposed to different concentrations of coix seed extract. The apoptosis was evaluated by light microscopy, Hoechst 33258 fluorescence staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and flow cytometry, respectively. The ultrastructural changes of PaTu-8988 cells were observed by transmission electron microscope. Results: The proliferative activity of PaTu-8988 cells pretreated with coix seed extract (10 μl/ml and 20 μl/ml) for 72 hours decreased in a dose-dependent manner. Light microscopy showed that the apoptotic cells became rounder and smaller, and were positively stained with fluorochrome Hoechst 33258 and TUNEL method. Flow cytometric analysis showed the appearance of a population of cells containing hypodiploid DNA in a time-dependent manner, the apoptosis rates induced by coix seed extract (20 μl/ml) were 7.1%±0.6% at 24 h, 11.3%±0.3% at 48 h and 15.1%±2.9% at 72 h, all higher than that of the control group (2.4%±1.1%, P<0.01). The early apoptotic change under transmission electron microscope was mainly the structural changes of mitochondria, then the classical ultrastructural feature of apoptosis, such as karyopyknosis, chromatic agglutination adjacent to the nuclear membrane and apoptotic body were found in late stage. Conclusions: The coix seed extract can induce apoptosis of human pancreatic cancer cells in a dose- and time-dependent manner, and the mitochondria might play a significant role in the early stage of apoptosis.
出处
《胃肠病学》
2005年第2期75-78,共4页
Chinese Journal of Gastroenterology
