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Objective:To study the variation and significance of Caspase-3 activity in the fructus bruceae emulsion inducing human leukemic cell (U937) apoptosis.Method:Inhibition of proliferation was measured by MTT assay. The cell apoptotic were examined by microscope, DNA fragmentation electrophoresis and flow cytometry (FCM). The activity of caspase-3 was detected by enzyme-linked immunosorbent assay.Result:The proliferation of U937 cells were significantly inhibited by 31.25~1000 μg/ml of fructus bruceae emulsion...

Objective:To study the variation and significance of Caspase-3 activity in the fructus bruceae emulsion inducing human leukemic cell (U937) apoptosis.Method:Inhibition of proliferation was measured by MTT assay. The cell apoptotic were examined by microscope, DNA fragmentation electrophoresis and flow cytometry (FCM). The activity of caspase-3 was detected by enzyme-linked immunosorbent assay.Result:The proliferation of U937 cells were significantly inhibited by 31.25~1000 μg/ml of fructus bruceae emulsion and showed a dose and time-dependent relationships. The IC50 value was 214 μg/ml. Typical morphological changes and DNA ladder were observed. while treated with fructus bruceae emulsion in125,250 and 500 μg/ml for 24 h,48 h and 72 h, the apoptotic ratios observed by FCM showed significant differences comparing with the control respectively (P< 0.01).The activity of caspase-3 was significant higher in the apoptosis than in control cells.Conclusion:Fructus bruceae emulsion could induce apoptosis of U937 cells in vitro, which may be one of the anticancer mechanisms.Caspase-3 activation mar participate in the processes of the apoptosis of U937 cells induced by fructus bruceae emulsion.

目的 :研究鸦胆子油乳诱导白血病细胞 (U937细胞 )凋亡及凋亡过程中caspase 3的变化及其意义。方法 :MTT法检测鸦胆子油乳对U937细胞增殖的抑制率 ;通过细胞形态、DNA片段化电泳、流式细胞术 (FCM )检测细胞凋亡 ;应用酶联反应法检测caspase 3活性。 结果 :在 31.2 5~ 1 0 0 0 .0 0mg/L浓度下鸦胆子油乳抑制U937细胞的增殖 ,IC5 0值为 2 14mg/L ,并呈现剂量 时间 效应关系 ;5 0 0mg/L鸦胆子油乳处理U937细胞 ,出现典型的细胞凋亡形态特征和DNA梯形带。流式细胞术检测显示 12 5、2 5 0、5 0 0mg/L鸦胆子油乳诱导U937细胞凋亡 ;在诱导细胞凋亡过程中Caspase 3活性明显增高 ,且随作用时间延长 ,活性增加。 结论 :鸦胆子油乳在体外能够诱导U937细胞凋亡 ,诱导细胞凋亡可能是鸦胆子油乳抗白血病作用的机制之一 ,caspase 3活化参与了鸦胆子油乳诱导U937细胞凋亡的调控。

Objective To explore the effect of Norcantharidin(NCTD)on growth and invasion of GBC-SD cell line of human gallhladder carcinoma.Methods GBC-SD cell line of human gallbladder carcinoma was cultivated in vivo.Six NCTD groups of different concentrations(6 wells in each group)were divided and a control group(n=6)was set up.The killer inhibitory rote,invasion motion capability,cell cycle and apoptosis rate of NCTD on GBC-SD cells were determined by MTr method,Matrigel experiment,the crossing-river experiment,and...

Objective To explore the effect of Norcantharidin(NCTD)on growth and invasion of GBC-SD cell line of human gallhladder carcinoma.Methods GBC-SD cell line of human gallbladder carcinoma was cultivated in vivo.Six NCTD groups of different concentrations(6 wells in each group)were divided and a control group(n=6)was set up.The killer inhibitory rote,invasion motion capability,cell cycle and apoptosis rate of NCTD on GBC-SD cells were determined by MTr method,Matrigel experiment,the crossing-river experiment,and FCM test.Results NCTD had obvious inhibitory effect on GBC-SD growth with the 50% inhibition concentration(IC_(50))being 56.18μg/ml.It inhibited GBC-SD invasion and migrated motion capability at the concentration of 5μg/ml.The inhibitory effect of NCTD increased,crossed cells of GBC-SD reduced,the dead crossed cells increased,and the crossing-river time obviously prolonged along with the increase of concentration(P<0.01),showing certain dose-time effect relationship.FCM analysis showed cells in G_2+M stage ob- viously increased in NCTD group,while they decreased in S stage with increased cell apoptosis.Conclusion NCTD not only inhibited the GBC-SD cell growth of human gallblader carcinoma,but also could inhibit its invasive capability under lower concentration.Its mechanism might be related to the inhibition of GBC-SD cell proliferation,interference growth cy- cle,inhibiting DNA synthesis,inducing cell apoptosis,and directly inhibiting the GBC-SD migration.

目的探讨去甲斑螯素(NCTD)对人胆囊癌 GBC-SD 细胞系生长、侵袭的影响。方法体外培养人胆囊癌 GBC-SD 细胞;实验分 NCTD 组(6个浓度梯度组,每组6孔)和对照组(n=6),分别用四唑盐比色法(MTT)、Matrigel 实验及过河实验、FCM 检测 NCTD 对 GBC-SD 细胞的杀伤抑制率、侵袭运动能力、细胞周期和凋亡率。结果 NCTD 对 GBC-SD 细胞生长有明显抑制作用,半数抑制浓度(IC_(50))为56.18μg/ml;在5μg/ml时即抑制 GBC-SD 细胞的体外侵袭和迁移运动能力;随浓度升高,NCTD 抑制作用增强,GBC-SD 过膜细胞减少、过膜细胞死亡增多,过河时间明显延长(P<0.01),呈剂量-时间效应关系。FCM 分析显示 NCTD 组 G_2+M 期细胞明显增多,S 期细胞减少,细胞凋亡率上升。结论 NCTD 不仅能抑制人胆囊癌 GBC-SD 细胞生长,亦可在低浓度下抑制其体外侵袭能力;机理可能与抑制 CBC-SD 细胞增殖,干扰生长周期,抑制 DNA 合成,诱导细胞凋亡和直接抑制 GBC-SD 细胞迁移运动有关。

 
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