甘肃猫儿眼甲基酯类化合物对人LGC-7910细胞毒性作用

Cytotoxic effects of methyl ester from mao er yan of Gansu origin on proliferation of human LGC-7910 strain cells

目的:观察甘肃猫儿眼甲基酯类化合物对人LGC-7910细胞毒性作用及其对细胞增殖的影响,为甘肃猫儿眼甲基酯类化合物的应用和开发提供实验依据。方法:实验于2006-06在甘肃政法学院公安分院实验中心完成。①体外培养人LGC-7910细胞48h,分为3组:甘肃猫儿眼甲基酯类化合物组:给予不同浓度(0.2,0.8和3.2mg/L)甘肃猫儿眼甲基酯类化合物;5-氟尿嘧啶组:给予10mg/L5-氟尿嘧啶;对照组:加等量的磷酸盐缓冲液。②用MTT法检测细胞生长抑制情况,用流式细胞仪测定细胞分裂周期中各期细胞数及凋亡率,用电镜扫描法观察细胞形态结构。③采用DUNNETT单因素方差分析对数据进行统计处理。结果:①MTT法测定甘肃猫儿眼甲基酯类化合物组(0.2,0.8,3.2mg/L)和5-氟尿嘧啶组对人LGC-7910细胞增殖抑制率分别为32.6%,65.7%,80.7%,46.5%,与对照组比较,差异有显著性(P<0.01),5-氟尿嘧啶组、3.2mg/L甘肃猫儿眼甲基酯类化合物组与0.8mg/L甘肃猫儿眼甲基酯类化合物组比较,差异有显著性(P<0.05)。②流式细胞仪测定,甘肃猫儿眼甲基酯类化合物组(0.2,0.8,3.2mg/L)和5-氟尿嘧啶组有61.6%,66.3%,70.6%,61.2%,滞留于G0G1期,在G2M期的细胞有11.9%,7.5%,4.8%,15.4%;凋亡率分别为11.6%,15.7%,26.2%,13.4%。③电镜扫描显示,对照组细胞膜完整,表面有很多微绒毛,甘肃猫儿眼甲基酯类化合物组细胞表面微绒毛断裂,出现孔洞,膜破裂,线粒体结构损伤,细胞出现凋亡、死亡。结论:甘肃猫儿眼甲基酯类化合物对人LGC-7910细胞具有明显的细胞毒作用,其机制与抑制分裂,破坏细胞形态和线粒体结构,诱导细胞凋亡有关。

AIM： To observe the eytotoxie effects and mechanism of methyl ester from mao er yan of Gansu origin on human LGC-7910 strain cells, so as to provide experimental evidence for the application and development of methyl ester from mao er yon of Gansu origin. METHDOS： The experiment was conducted at the Experimental Center of Public Security, Gansu Political Science and Law Institute in June 2006. （1）LGC-7910 strain cells were cultured in vitro for 48 hours and divided into 3 groups： methyl ester group, which was given methyl ester at different at doses （0.2, 0.8 and 3.2 mg/L）; 5-fluorouraeil （5-FU） group, which was given 10 mg/L 5-FU; control group, which was given matching phosphate buffer. （2）The cell growth inhibition was detected with MTT method; the number of cells and apoptotic rate were measured by flow eytometry. （3） DUNNETT ANOVA was used to handle the data. RESULTS： （1）MTT detection showed that the inhibitory rate of the methyl ester group （0.2, 0,8 and 3.2 mg/L） and 5-FU group were 32.6%,65.7%, 80.7% and 46.5%, which significantly differed from the control group （P 〈 0.01）; there were significant differences among the 5-FU group, 3.2 mg/L methyl ester group and 0.8 mg/L methyl ester group （P 〈 0.05）. （2）Flow cytometry results showed that the cells of G0G1 phase in the the methyl ester group （0.2,0.8,3.2 mg/L） and 5-FU group were 61.6%,66.3% 70.6% and 61.2%,those of G2M phase were 11.9%,7.5%,4.8% and 15.4%,the apoptosls rate was 11.6% and 15.7%,26.2% and 13.4%. （3） Electron microscope showed that the cells of the control group had complete cell membrane with a great amount of microvilli; the microvilli of the cells in the methyl ester group were collapsed with holes, the membrane was broke; the mitochondria was injured, and cell apoptosis and death were found. CONCLUSION： Methyl ester from mao er yon of Gansu origin has significant cytotoxic effects on human LGC-7910 strain cells and the mechanism may relate to inhibiting cell division, destroying the structure of cell membrane and mitochondria, and inducing cell apoptosis.