冬凌草甲素诱导HL-60细胞凋亡

目的　研究冬凌草甲素诱导人白血病HL 6 0细胞凋亡的作用。方法　形态学观察 ,DNA凝胶电泳及流式细胞术。结果　冬凌草甲素能显著地诱导HL 6 0细胞发生凋亡 ,其作用呈明显的浓度效应关系和时间依赖性。形态学观察可见凋亡小体的形成 ,琼脂糖凝胶电泳可见明显的DNA梯带 ;流式细胞仪检测到G1亚峰。结论　冬凌草甲素能诱导HL 6 0细胞凋亡 ,并与其细胞杀伤活性相互平行 ,提示冬凌草甲素的抗癌活性与诱导肿瘤细胞凋亡相关     

白英水提物诱导人宫颈癌HeLa细胞凋亡的实验研究

目的:观察白英水提物诱导人宫颈癌HeLa细胞凋亡的作用,并初步探讨其分子机制。方法:常规方法制备白英水提物,体外培养人宫颈癌HeLa细胞。实验组设白英水提物12.5、25.0、50.0mg/mL3种浓度,以顺铂(25.0mg/L)为阳性对照组。采用细胞毒试验(MTT法)检测HeLa细胞抑制率,荧光显微镜观察HeLa细胞形态变化,琼脂糖凝胶电泳检测DNA梯形泳带,半定量RT-PCR法检测凋亡相关基因survivin mRNA和caspase-3mRNA表达的变化。结果:白英水提物对HeLa细胞增殖有抑制作用,且呈明显的时效和量效关系,并诱导其发生凋亡,表现为细胞核固缩、染色质凝集、边聚等。琼脂糖凝胶电泳显示DNA梯形泳带,半定量RT-PCR法检测显示survivin mRNA表达下调、caspase-3mRNA表达上调。结论:白英水提物对HeLa细胞有较强的增殖抑制和诱导凋亡作用,其诱导HeLa细胞凋亡的分子机制可能与上调caspase-3基因表达、下调survivin基因表达有关。     

高三尖杉酯碱对HL-60细胞端粒酶量的抑制效应

目的　研究高三尖杉酯碱 (HHT)对HL 6 0细胞端粒酶活性的影响及诱导凋亡作用。方法　采用端粒重复序列扩增法 (TRAP) 酶联免疫吸附试验(ELISA)检测了HL 6 0细胞的端粒酶量变化 ,用细胞形态学观察 ,DNA琼脂糖凝胶电泳 ,流式细胞术检测和DNA片段原位末端标记 (TUNEL)法检测了细胞凋亡现象。结果　 5～ 5 0 0 μg·L- 1HHT处理HL 6 0细胞 0～ 4 8h ,HL 6 0细胞端粒酶量呈剂量依赖性和时间依赖性下降。同时 ,HL 6 0细胞发生了凋亡。结论　HHT有降低HL 6 0细胞的端粒酶量 ,诱导细胞凋亡作用。     

金喜素诱导HL-60细胞凋亡依赖caspase活化

目的　探讨金喜素诱导急性白血病细胞凋亡的生化机制。方法　采用MTT法测定金喜素对HL 6 0细胞的杀伤作用 ;通过形态学、DNA凝胶电泳及AnnexinVFITC染色 ,研究金喜素对靶细胞的促凋亡活性 ;采用caspase 8、3特异性抑制剂IETD fmk、DEVD CHO ,分析金喜素介导的靶细胞凋亡与caspase活化的关系。结果　经 0 1μmol·L-1金喜素处理至 8、12及 16h时 ,HL 6 0细胞的存活率依次降至6 2 %± 12 %、、43%± 15 %及 32 %± 10 % ,低于对照 (P <0 0 5 ) ,同时 ,靶细胞逐渐出现磷脂酰丝氨酸外化 ,并产生梯状DNA及凋亡小体 ;经caspase抑制剂与金喜素联合处理12、16h时 ,HL 6 0细胞的存活率高于单用金喜素组 ,分别为77%± 14%、6 5 %± 16 % (IETD fmk组 )与 74%± 12 %、6 0 %± 11% (DEVD CHO组 ) ,同时 ,靶细胞的梯状DNA与凋亡小体的诱生也明显受抑。结论　金喜素对HL 6 0细胞具有较强的促凋亡作用 ,该过程可能依赖caspase 8、caspase 3的活化     

中华眼镜蛇毒血清抑制HL60细胞增殖及诱导凋亡的实验研究

目的　探讨中华眼镜蛇毒血清体外对白血病细胞系(HL6 0 )的抑制作用及其机制。方法　以人粒细胞白血病细胞系 (HL6 0 )为靶细胞 ,观察口服中华眼镜蛇毒血清对肿瘤细胞增殖的影响 ,并采用DNA片断凝胶电泳、流式细胞仪分析药物体外诱导细胞凋亡情况。结果　中华眼镜蛇毒血清在体外显著抑制HL6 0细胞的生长 ,其抑制作用随着剂量的加大而增强 ,高剂量 (7 5g·L-1)的抑制效果相近于阿霉素(5mg·L-1) ,和空白对照组比较差异有显著性。经DNA电泳、流式细胞仪分析显示中华眼镜蛇毒血清与HL6 0细胞共同培养可诱导其发生细胞凋亡。结论　中华眼镜蛇毒血清体外能显著抑制HL6 0细胞的生长 ,此作用可能与诱导白血病细胞发生凋亡有密切关系。     

青蒿琥酯诱导人早幼粒白血病细胞HL60凋亡的线粒体机制

目的 :探讨青蒿琥酯 (artesunate ,Art)诱导人急性早幼粒白血病细胞HL6 0凋亡的线粒体机制。方法 :用Art处理HL6 0细胞 ,通过MTT比色法检测细胞增殖抑制的效果 ;透射电镜、DNA凝胶电泳技术观察细胞的凋亡 ;流式细胞术 (flowcytometry ,FCM)进行细胞周期分析和线粒体膜电位 (mitochon drialmembranepotential ,MMP)水平的检测 ;比色法检测半胱 天冬氨酸蛋白酶 3(cysteinecontainingas partate,Caspase 3)活性。结果 :Art呈浓度和时间依赖性的抑制HL6 0细胞的增殖 (P <0 .0 5 )。Art诱导后HL6 0细胞出现典型的凋亡形态学变化和生化改变 ,G2 M期细胞比例增高 ,S期减少 ,凋亡率上升 (P <0 .0 5 ) ,线粒体膜电位降低 (P <0 .0 5 ) ,Caspase 3活性增强 (P <0 .0 5 )。结论 :Art可能通过线粒体 半胱 天冬氨酸蛋白酶 3特定途径诱导HL6 0细胞凋亡。     

梁金菇多糖促人急性早幼粒细胞白血病细胞系(HL-60)细胞凋亡研究

目的 :观察梁金菇多糖对人类早幼粒细胞白血病细胞系 (HL 6 0 )是否具有凋亡诱导作用 ,并进一步研究半胱天冬酶 3(Caspase 3)基因在此过程中的作用。方法 :四氮唑蓝比色法 (MTT法 )观察梁金菇多糖对HL 6 0细胞的抑制率 ,应用形态学、流式细胞术、DNA凝胶电泳等方法检测细胞凋亡的发生。应用半定量逆转录 聚合酶链反应 (RT PCR)法检测凋亡相关基因Caspase 3mRNA表达的变化。结果 :梁金菇多糖对HL 6 0细胞的生长有明显的抑制作用 ,并诱导肿瘤细胞发生凋亡 ;在HL 6 0细胞凋亡过程中 ,凋亡相关基因Caspase 3转录水平比用药前增强。结论 :梁金菇多糖促HL 6 0细胞凋亡 ,且与Caspase 3基因表达有关     

聚酯型儿茶素诱导HL-60细胞凋亡

目的　探讨聚酯型儿茶素的抗癌作用机制。方法　应用MTT法、电镜、琼脂糖凝胶电泳、流式细胞术、RT PCR和免疫细胞化学等方法研究细胞凋亡及其机制。结果　聚酯型儿茶素作用于HL 6 0细胞后 ,扫描和透射电镜观察到异染色质固缩、边集及凋亡小体形成 ;DNA琼脂糖凝胶电泳上可见典型的“阶梯状”条带 (DNALadder) ;AnnexinV FITC和PI双染后流式细胞术分析显示 ,在一定时间一定剂量范围内 ,聚酯型儿茶素诱导细胞凋亡的作用呈现浓度和时间依赖性增强的趋势 ,但是大剂量或长时间将主要导致细胞坏死 ;聚酯型儿茶素作用于HL 6 0细胞 2 4h后 ,细胞bcl 2基因及蛋白产物表达明显降低。结论　聚酯型儿茶素可诱导HL 6 0细胞调亡 ,诱导肿瘤细胞凋亡可能是聚酯型儿茶素抗肿瘤作用的重要机制     

中华眼镜蛇毒诱导HL-60细胞凋亡及其机制探讨

目的　观察中华眼镜蛇毒体外对白血病细胞凋亡的影响 ,并探讨其可能的作用机制。方法　本研究采用血清药理学方法 ,以人白血病细胞HL 60细胞为靶细胞 ,通过流式细胞仪分析、DNA片段凝胶电泳、免疫组织化学 (S P法 )观察中华眼镜蛇毒兔血清体外对细胞凋亡的诱导作用及癌基因表达的影响。结果　经DNA电泳、流式细胞仪分析显示 :中华眼镜蛇毒兔血清与HL 60细胞共同培养 48h可诱导其细胞凋亡增加 ,bcl 2、c myc基因蛋白表达下调。结论　中华眼镜蛇毒兔血清体外抑制HL 60细胞增殖可能与其诱导细胞凋亡及使细胞bcl 2、c myc基因蛋白表达下调密切相关     

二烯丙基二硫诱导人白血病HL-60细胞凋亡及机制

目的　探讨二烯丙基二硫 (DADS)诱导人白血病HL 6 0细胞凋亡的生物学效应及抗肿瘤机制。方法　通过MTT还原法检测DADS对该细胞系生长的影响 ;用电镜、荧光显微镜、流式细胞仪和细胞凋亡原位检测 (TUNEL)研究DADS诱导的细胞凋亡。SP免疫组化法检测细胞内Bcl 2、Bax蛋白的表达。结果　DADS能抑制HL 6 0细胞的生长 ,DADS处理HL 6 0细胞后 ,电镜下细胞呈凋亡特征性改变 :体积变小 ,核浓缩 ,凋亡小体形成 ,流式细胞仪示不同浓度DADS作用于HL 6 0细胞 ,亚G1期细胞明显增多 ,TUNEL测凋亡指数增加。SP免疫组化结果表明 10mg·L-1DADS处理细胞 2 4h后 ,Bax蛋白表达升高 ,Bcl 2蛋白表达下降。结论　DADS有诱导人白血病HL 6 0细胞凋亡的作用 ,其机制与Bcl 2 /Bax比例下降有关。     

一叶秋碱诱导人白血病HL—60细胞凋亡

目的 研究一叶秋碱能否诱导ＨＬ－６０细胞凋亡,方法 用ＭＴＴ法检测一叶秋碱对细胞增殖影响;应用流式细胞仪检测凋亡细胞数;采用琼脂糖凝胶电泳法观测ＤＮＡ碎片,透射电镜观察凋亡的形态改变,结果：一叶秋碱５－８０ｍｇ．Ｌ＾－１能诱导ＨＬ－６０细胞凋亡,电镜观察到典型的凋亡形态学改变,电泳呈现出阶梯状条带,流式细胞仪检测到凋亡率随剂量的增高而升高,ＭＴＴ法示一叶秋碱抑制ＨＬ－６０细胞增殖,并且呈时间,剂量     

Tartary buckwheat flavonoid activates caspase 3 and induces HL-60 cell apoptosis

It has been proposed that flavonoids may have potential as anticancer agents. In this study, we showed that tartary buckwheat flavonoid (TBF) obviously inhibits the growth of human acute myelogenous leukemia (AML) HL-60 cells by MTT assay. The inhibitory effect of TBF on the proliferation of HL-60 cells is related to the induction of apoptosis, which is confirmed by DNA ladder formation on gel electrophoresis and apoptosis morphological changes under light microscope. Furthermore, HL-60 cells undergo rapid apoptosis upon treatment with TBF, as indicated by increased annexin V binding capacity and caspase 3 activation with flow cytometric analysis. Thus, our data provide a potential mechanism for the chemopreventive activity of tartary buckwheat flavonoid and suggest that it may have a potentially therapeutic role for human leukemia.     

三丁酸甘油酯对白血病细胞株HL-60体外作用的研究

目的探讨三丁酸甘油酯(TB)对白血病细胞株HL-60细胞增殖的抑制作用,并对其作用机制进行初步探讨。方法采用MTT法观察细胞增殖变化。应用免疫组化检测抑癌基因P16的表达。应用流式细胞仪观察细胞周期的改变。通过DNA电泳观察细胞凋亡。结果TB可以抑制细胞增殖。1.0 mmol/L的TB作用72 h,有典型的DNA梯形条带。细胞周期阻滞在G0/G1期。结论TB能抑制HL-60细胞增殖,诱导细胞凋亡,其机制可能与上调P16表达有关。     

Baicalein induced in vitro apoptosis undergo caspases activity in human promyelocytic leukemia HL-60 cells.

In this study, we evaluated the potential apoptosis effects of baicalein on human promyelocytic leukemia HL-60 cells in vitro. Apoptosis induction, cell viability, morphology and caspase-3 activity were then performed to determine by flow cytometric assay, DNA gel electrophoresis, anti-ADP-ribose stain and determination of caspase-3 activity. There is a significant difference in cell death of HL-60 cells that was detected between baicalein-treated and untreated groups. Furthermore, there was a further significant increase in apoptosis induction when cells were treated with baicalein compared to without baicalein treated groups. Flow cytometric assays and DNA fragmentation gel electrophoresis also confirmed baicalein induced apoptosis in HL-60 cells. Baicalein also promoted caspase-3 activity then leading to cleavage of poly-ADP-ribose polymerase finally leading to DNA fragmentation occurrence. Furthermore, the baicalein-induced apoptosis was markedly blocked by the broad-spectrum caspase inhibitor, z-VAD-fmk. Taken together, these results suggest that treatment of human leukemia HL-60 cells with baicalein induced apoptosis through activation of caspase-3 activity.     

Diallyl disulfide induces apoptosis in human leukemia HL-60 cells through activation of JNK mediated by reactive oxygen

Diallyl disulfide (DADS) is a chemopreventive agent that can induce apoptosis in many tumor cells. Reactive oxygen species (ROS) are important mediators in apoptosis induced by various stimuli, including chemopreventive agents. The phosphotransferase c-JUN N-terminal kinase (JNK) has been shown to regulate apoptosis. In this study, we found that DADS-induced apoptosis in human leukemia HL-60 cells is mediated by ROS-activated JNK. The DADS-treated HL-60 cells showed a dose- and time-dependent decrease in cell viability and proliferation. Agarose gel electrophoresis of cells treated with 10.0 or 20.0 mg/L DADS for 24 h showed a characteristic ladder pattern in their DNA. Levels of DADS-induced ROS, as measured by 2',7'-dichlorofluorescein diacetate (DCFH-DA) fluorescence, also showed dose- and time-dependent increases in HL-60 cells. Activity of JNK was induced by DADS in a dose-dependent manner; HL-60 cells exposed to 10.0 mg/L DADS for 8 h showed maximum levels of phosphorylated JNK, which decreased when exposed for additional 4h. In contrast, Sp600125, a specific inhibitor of JNK, blocked apoptosis of HL-60 cells exposed to DADS. N-Acetylcysteine (NAC), a known antioxidant, also decreased ROS generation, effectively blocked apoptosis, and decreased DADS-induced phosphorylated JNK levels. These results suggest that JNK is involved in DADS-induced ROS-mediated apoptosis in HL-60 cells.     

异三尖杉酯碱诱导HL-60细胞凋亡

用透射电镜观察、DNA凝胶电泳及流式细胞术研究了异三尖杉酯碱(IHT)诱导HL-60细胞凋亡的作用。结果证明异三尖杉酯碱能快速、显著地诱导HL-60细胞发生凋亡,其作用呈明显的浓度-效应关系和时间依赖性。IHT处理的细胞出现凋亡相关的特征性变化。在电镜观测中发现细胞核内染色质浓缩边集、核碎裂及凋亡小体形成;琼脂糖凝胶电泳可见明显的DNA梯;流式细胞光度术出现显著的G1亚峰。经10^-7mol·L^-1IHT处理120min,可使43.8%HL-60细胞发生凋亡。IHT有明显诱导HL-60细胞凋亡的作用,并与其细胞杀伤活性相平行,提示IHT的抗癌活性与诱导肿瘤细胞凋亡相关。这些实验结果对进一步研究、开发IHT有重要实用价值。     

金荞麦Fr4诱导HL-60细胞凋亡及对端粒酶活性的影响

目的研究金荞麦有效部位Fr4能否诱导HL-60细胞凋亡及对端粒酶活性的影响。方法用MTT法检测Fr4对细胞增殖的影响;光学显微镜和透射电镜观察细胞形态改变;Annexin-V/PI双染法检测细胞膜介导的凋亡;琼脂糖凝胶电泳观察DNA碎片;TRAP-PCR-ELISA检测端粒酶活性。结果金荞麦Fr4 60~240 mg.L-1能诱导HL-60细胞凋亡,电镜观察到典型的凋亡细胞,电泳呈现出阶梯状条带,流式细胞仪检测到早期凋亡细胞数随剂量的增加而升高。MTT法示金荞麦Fr4抑制HL-60细胞增殖,并且呈剂量依赖性趋势,药物作用24 h的IC50为115 mg.L-1,Fr4诱导HL-60细胞凋亡过程中,端粒酶活性下降。结论金荞麦Fr4可诱导HL-60细胞凋亡,其机制可能与端粒酶活力下降有关。     

Arsenic induced apoptosis in rat liver following repeated 60 days exposure

BACKGROUND: Accumulation of the wide spread environmental toxin arsenic in liver results in hepatotoxcity. Exposure to arsenite and other arsenicals has been previously shown to induce apoptosis in certain tumor cell lines at low (1-3 microM) concentration. AIM: The present study was focused to elucidate the role of free radicals in arsenic toxicity and to investigate the nature of in vivo sodium arsenite induced cell death in liver. METHODS: Male wistar rats were exposed to arsenite at three different doses of 0.05, 2.5 and 5mg/l for 60 days. Oxidative stress in liver was measured by estimating pro-oxidant and antioxidant activity in liver. Histopathological examination of liver was carried out by light and transmission electron microscopy. Analysis of DNA fragmentation by gel electrophoresis was used to identify apoptosis after the exposure. Terminal deoxy-nucleotidyl transferase mediated dUTP Nick end-labeling (TUNEL) assay was used to qualify and quantify apoptosis. RESULTS: A significant increase in cytochrome-P450 and lipid peroxidation accompanied with a significant alteration in the activity of many of the antioxidants was observed, all suggestive of arsenic induced oxidative stress. Histopathological examination under light and transmission electron microscope suggested a combination of ongoing necrosis and apoptosis. DNA-TUNEL showed an increase in apoptotic cells in liver. Agarose gel electrophoresis of DNA of hepatocytes resulted in a characteristic ladder pattern. CONCLUSION: Chronic arsenic administration induces a specific pattern of apoptosis called post-mitotic apoptosis.