二烯丙基二硫对人白血病HL-60细胞株survivin表达的影响

目的:观察二烯丙基二硫(diallyl disulfide,DADS)作用于人白血病HL-60细胞株后,细胞增殖及survivin蛋白表达的变化,探讨DADS对HL-60细胞增殖的影响及其机制。方法:DADS作用HL-60后,MTT法检测细胞增殖情况,SP-免疫组化法检测细胞survivin蛋白表达的变化。结果:DADS可以呈时间-浓度依赖性抑制HL-60细胞增殖并下调survivin蛋白的表达。结论:DADS能通过下调survivin蛋白的表达而抑制HL-60细胞的增殖。     

Inhibition of ERK and activation of p38 are involved in diallyl disulfide induced apoptosis of leukemia HL-60 cells

We investigated the effects of diallyl disulfide (DADS) on the induction of apoptosis in human Leukemia cell line HL-60 and explored the roles of mitogen-activated protein kinase (ERK and p38 MAPK) pathways in the growth inhibition and apoptosis induced by DADS. MTT assay was used to determine the DADS induced cell growth inhibition in HL-60 cells. Flow cytometry and DNA fragmentation were used to examine the roles of apoptosis in DADS-mediated cell death. Western blot analysis of the expression of phospho-MAPKs (ERK and p38) was employed to elucidate the possible mechanisms of DADS induced apoptosis. We found that growth inhibition of HL-60 cells treated with DADS exhibited a dose-dependent response (P<0.05) and DADS induced significant apoptosis. DADS at the concentration of 10 mg/L persistently activated p38 and simultaneously reduced ERK activity. PD98059, an inhibitor of ERK upstream activators MAPK kinase MKK1 and MKK2, promoted cytotoxicity and apoptosis in HL-60 cells treated with DADS. In contrast, SB203580, an inhibitor of p38, decreased cytotoxicity and apoptosis induced by DADS. Therefore, DADS can effectively inhibit the proliferation and induce apoptosis of human leukemia cell line HL-60. Inhibition of ERK signaling pathways and activation of p38 signaling pathways are likely involved in DADS induced apoptosis in HL-60 cells.     

Chromene induces apoptosis via caspase-3 activation in human leukemia HL-60 cells

In this study, the potent anti-tumor effects of brown algae on human leukemia HL-60 cells were investigated. The Sargassum siliquastrum extract among the 14 species of brown algae exhibited profound growth inhibitory effect on HL-60 cells in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, therefore, S. siliquastrum was selected for use in further experiments. The highest inhibitory activity of S. siliquastrum on HL-60 cells was detected in the chloroform fraction, and the active compound was identified as a kind of chromene, sargachromanol E (SE). SE treatment showed significant growth inhibitory effects on HL-60 cells in a dose-dependent manner by inducing apoptosis, as evidenced by the formation of apoptotic bodies, fragmented DNA ladder, and the accumulation of DNA in the sub-G₁ phase of cell cycle. SE induced apoptosis was accompanied by downregulation of Bcl-xL, upregulation of Bax, activation of caspase-3, and cleavage of poly (ADP-ribose) polymerase (PARP). Moreover, z-DEVD-fmk, a caspase-3 inhibitor, significantly inhibited cell cytotoxicity, apoptotic characteristics such as apoptotic bodies, sub-G₁ DNA content, and cleavage of PARP induced by SE. These results suggest that SE exerts its growth inhibitory effects on HL-60 cells through caspase-3-mediated induction of apoptosis. Therefore, SE offers promising chemotherapeuric potential to prevent cancers such as human leukemia.     

Induction of apoptosis by penta-O-galloyl-beta-D-glucose through activation of caspase-3 in human leukemia HL-60 cells.

Penta-O-galloyl-beta-D-glucose is structurally related to (-)-epigallocatechin gallate and is isolated from hydrolyzed tannin. Penta-O-galloyl-beta-D-glucose can inhibit tumor promotion by teleocidin. We investigated the effects of penta-O-galloyl-beta-D-glucose and various tea polyphenols on cell viability in human leukemia HL-60 cells. In this study, we demonstrated that penta-O-galloyl-beta-D-glucose was able to induce apoptosis in a concentration- and time-dependent manner; however, other polyphenols were less effective. We further investigated the molecular mechanisms of penta-O-galloyl-beta-D-glucose-induced apoptosis. Treatment with penta-O-galloyl-beta-D-glucose caused induction of caspase-3/CPP32 activity in dose- and time-dependent manner, but not caspase-1 activity, and induced the degradation of poly-(ADP-ribose) polymerase. Pretreatment with acetyl-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO) and Z-Val-Ala-Asp-fluoromethyl-ketone (Z-VAD-FMK) inhibited penta-O-galloyl-beta-D-glucose-induced DNA fragmentation. Furthermore, treatment with penta-O-galloyl-beta-D-glucose (50 microM) caused a rapid loss of mitochondrial transmembrane potential, release of mitochondrial cytochrome c into cytosol, and subsequent induction of procaspase-9 processing. Our results indicate that penta-O-galloyl-beta-D-glucose allows caspase-activated deoxyribonuclease to enter the nucleus and degrade chromosomal DNA, and induces DFF-45 (DNA fragmentation factor) degradation. These results lead to a working hypothesis that penta-O-galloyl-beta-D-glucose-induced apoptosis is triggered by the release of cytochrome c into the cytosol, procaspase-9 processing, activation of caspase-3, degradation of poly-(ADP-ribose) polymerase, and DNA fragmentation caused by the caspase-activated deoxyribonuclease through the digestion of DFF-45. The induction of apoptosis by penta-O-galloyl-beta-D-glucose may provide a pivotal mechanism for its cancer chemopreventive action.     

Glaucocalyxin A induces apoptosis in human leukemia HL-60 cells through mitochondria-mediated death pathway

Glaucocalyxin A (GLA) is a biologically active ent-kauranoid diterpenoid isolated from Rabdosia japonica var. glaucocalyx, a traditional Chinese medicinal herb, which has been shown to inhibit tumor cell proliferation. However, the mechanism underlying GLA-induced cytotoxicity remains unclear. In this study, we focused on the effect of GLA induction on apoptosis, the mitochondria-mediated death pathway and the accumulation of reactive oxygen species (ROS) in human leukemia cells (HL-60). GLA could induce a dose-dependent apoptosis in HL-60 cells as characterized by cell morphology, DNA fragmentation, activation of caspase-3, -9 and an increased expression ratio of Bax/Bcl-2. The mitochondrial membrane potential (Δψ_m) loss and cytochrome c release from mitochondria to cytosol were observed during the induction. Moreover, GLA caused a time- and dose-dependent elevation of intracellular ROS level in HL-60 cells, and N-acetyl-l-cysteine (NAC, a well-known antioxidant) could block GLA-induced ROS generation and apoptosis. These data suggest that GLA induces apoptosis in HL-60 cells through ROS-dependent mitochondrial dysfunction pathway.     

二烯丙基二硫诱导人白血病细胞系HL-60细胞分化的实验研究

目的　探讨二烯丙基二硫 (DADS)对人急性髓性白血病细胞系HL 6 0细胞增殖抑制及诱导分化作用的影响。方法　采用MTT法检测对细胞增殖的影响 ,通过观察细胞形态、硝基四氮唑蓝 (NBT)还原反应、流式细胞仪测定鉴定细胞分化。结果　HL 6 0细胞经DADS处理后 ,细胞增殖受抑 ,呈剂量效应关系。 0 6 2 5～ 1 2 5mg·L-1时NBT还原反应增强 (P <0 0 1或 <0 0 5 ) ,流式细胞术显示可诱导表面分化抗原CD11b表达升高及细胞周期阻滞于G1期。结论　小剂量DADS长时间作用对HL 6 0细胞具有诱导分化作用 ,其作用相当于全反式维甲酸 ,对于白血病的治疗具有潜在的应用价值     

华蟾素诱导HL-60细胞凋亡及机制研究

目的：观察华蟾素（cinobufacini）对白血病HL-60细胞的增殖抑制作用和诱导凋亡作用的机制。方法：以HL-60细胞为研究对象，采用MTT法观察细胞增殖作用；Annexin V-FITC／PI双染和吖啶橙／溴乙锭（AO／EB）荧光染色法检测细胞凋亡；罗丹明染色法检测线粒体膜电位；分光光度法检测Caspase-3活性。结果：在0．78～6．25μg&#183;ml^-1，华蟾素能明显的抑制HL-60细胞的增殖；华蟾素作用24h后，AnnexinV阳性的细胞从7．81％增加至66．02％，而且在荧光显微镜下可以明显观察到凋亡细胞；线粒体膜电位下降和Caspase-3活性升高。结论：华蟾素能够抑制HL-60细胞增殖，这种作用与其诱导细胞凋亡破坏线粒体功能和激活Caspase-3有关。     

Dracorhodin perchlorate induces apoptosis in HL-60 cells

Dracorhodin perchlorate, an anthocyanin red pigment, induces human premyelocytic leukemia HL-60 cell death through apoptotic pathway. Caspase -1, -3, -8, -9, and -10 inhibitors partially reversed the cell death induced by dracorhodin perchlorate. Caspase-3 and -8 were activated followed to the degradation of caspase-3 substrates, inhibitor of caspase-activated DNase (ICAD) and poly-(ADP-ribose) polymerase (PARP). Dracorhodin perchlorate up-regulated the expression ratio of mitochondrial proteins, Bax/Bcl-X_L. The cell death was accompanied with phosphorylation of ERK, JNK and p38 MAPK and partially reduced by MEK inhibitor (PD98059), JNK MAPK inhibitor (SP600125) and p38 MAPK inhibitor (SB 203580). Taken together, dracorhodin perchlorate-induced apoptosis in HL-60 cells via up-regulation of Bax, activation of caspases and ERK/p38/JNK MAPKs.     

Induction of apoptosis by Cordyceps militaris through activation of caspase-3 in leukemia HL-60 cells

Cordyceps militaris is a traditional herbal ingredient frequently used for tonic and medicinal purposes in eastern Asia. The hot water extract of its cultivated fruiting bodies demonstrated a potent cytotoxic effect against the proliferation of the human premyelocytic leukemia cell HL-60, with an IC50 of 0.8 mg/ml for a 12-h treatment. It induced the characteristic apoptotic symptoms in the HL-60 cells, including DNA fragmentation and chromatin condensation, occurring within 12-16 h of treatment at a dose of 1 mg/ml. The activation of caspase-3 and the specific proteolytic cleavage of poly (ADP-ribose) polymerase were detected during the course of apoptosis induction. These results indicate that the hot water extract of Cordyceps militaris fruiting bodies inhibited cancer cell proliferation by inducing cell apoptosis through the activation of caspase-3, and that the Cordyceps militaris extract may therefore have therapeutic potential against human leukemia.     

毛兰素诱导人白血病HL—60细胞的凋亡

目的：研究毛兰素对HL－60细胞增殖的抑制作用,探讨其诱导细胞凋亡的机制。方法：用MTT比色法测定了毛兰素对HL－60细胞增殖的抑制作用：应用荧光显微镜、透射电镜、DNA电泳及流式细胞仪观察了药物对细胞凋亡的诱导作用,并用免疫组化的方法从基因水平阐述了凋亡的发生。结果：毛兰素20－81．9nmol/L在72h内显著抑制HL－60细胞增殖,作用24h后,对HL－60细胞的IC50为38nmol/L,而阳性对照药长春新碱对HL－60细胞的IC50为101nmol/L,前者明显优于后者;形态学观察可见凋亡的特征性改变;琼脂糖电泳出现典型的DNA“ladder”;流式细胞仪结果表明细胞被阻滞于G2／M期;免疫组化可见bcl-2表达下降,bax表达升高。结论：毛兰素显著抑制HL－60细胞的生长,该抑制作用可能是通过诱导细胞凋亡和改变HL－60细胞bcl-2和bax基因的表达而实现的。     

小剂量二烯丙基二硫联合全反式维甲酸对HL-60细胞的生长抑制和诱导分化作用研究

二烯丙基二硫(diallyl disulfide,DADS)具有抑制肿瘤细胞增殖,调控细胞周期依赖素激酶、信号转导,诱导肿瘤细胞分化、凋亡及影响癌基因与抑癌基因表达的作用。小剂量DADS(1·25mg·L-1)可以抑制JAK1/STAT3信号通路,将HL-60细胞阻滞在G0/G1期,诱导HL-60细胞向粒系分化,其作用与全反式维甲酸(ATRA)相当[1~3]。本实验将探讨小剂量DADS与ATRA联合应用对HL-60细胞的生长抑制及诱导分化效应。1材料和方法参见文献2。2结果2.1DADS与ATRA单独或联合用药对HL-60细胞生长的影响1·25mg·L-1DADS或ATRA对体外培养的HL-60细胞均…     

消瘤平体外诱导人白血病HL-60细胞凋亡及机制研究

目的研究消瘤平对人白血病细胞增殖的抑制及诱导凋亡的作用,揭示其抗白血病的部分作用机制。方法体外常规培养HL-60细胞,以不同浓度的消瘤平作用于细胞,MTT法检测细胞生长抑制率;流式细胞术检测凋亡率和细胞周期;ELISA法检测培养上清液中Caspase-3、Fas和Bcl-2的含量。结果与空白对照组相比,作用24 h后,消瘤平显著抑制HL-60细胞的增殖,且呈时间和剂量依赖性;提高HL-60细胞的凋亡率,并诱导出现G2-M细胞周期阻滞;消瘤平作用48 h后,Caspase-9和Fas蛋白表达量增加,Bcl-2表达减少。结论消瘤平抑制人白血病HL-60细胞的作用机制与阻滞细胞周期于G2-M期和诱导细胞凋亡有关,其诱导凋亡作用可能通过上调Caspase-9、Fas和下调Bcl-2而实现。     

槲皮素诱导人白血病HL-60细胞凋亡(英文)

目的:研究槲皮素是否能诱导人白血病HL-60细胞凋亡.方法:应用琼脂糖凝胶电泳法观察DNA碎片;采用流式细胞仪检测DNA断裂;电镜技术观察凋亡的形态学改变,用MTT测定法测定细胞增殖.结果:槲皮素15-120 μmol·L~(-1)诱导HL-60细胞凋亡,电镜观察到典型的形态学改变,电泳显示梯状条带,槲皮素能剂量依赖性地触发DNA降解及抑制细胞增生(IG_(50)和95％可信区间分别为43(30-61)μmol·L~(-1).结论:槲皮素诱导人白血病HL-60细胞凋亡.     

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。     

Fluoride induces apoptosis by caspase-3 activation in human leukemia HL-60 cells

Even though fluoride toxicity is increasingly being considered to be important, very little information is available on the mechanism of action of fluoride. In the present study, the toxicity of fluoride on human leukemia (HL-60) cells was investigated and the involvement of caspase-3 was also studied. Fluoride induced apoptosis in HL-60 cells in a dose- and time-dependent manner. Annexin staining and DNA ladder formation on agarose gel electrophoresis further revealed that HL-60 cells underwent apoptosis on exposure to 2-5 mM fluoride. Western blotting using polyclonal anti-caspase-3 antibody and mouse anti-human poly(ADP-ribose) polymerase (PARP) monoclonal antibody was performed to investigate caspase-3 and PARP activity. Fluoride led to the activation of caspase-3 which was evident by the loss of the 32 kDa precursor and appearance of the 17 kDa subunit. Furthermore, intact 116 kDa PARP was cleaved by fluoride treatment as shown by the appearance of a cleaved 89 kDa fragment. The results clearly suggest that fluoride causes cell death in HL-60 cells by causing the activation of caspase-3 which in turn cleaves PARP leading to DNA damage and ultimately cell death.     

Induction of apoptosis and transient increase of phosphorylated MAPKs by diallyl disulfide treatment in human nasopharyngeal carcinoma CNE2 cells

This study was undertaken to elucidate the effect of diallyl disulfide (DADS), an oil-soluble organosulfur compound found in garlic, in suppressing human nasopharyngeal carcinoma cells. A potent increase (of at least 9-fold) in apoptotic cells has accompanied 1) a decrease in cell viability, 2) a increase of the fraction of S-phase cells by up to 63.8%, and 3) a transient increase of the phospho-p38 and phospho-p42/44 (phosphorylated p38 MAPK and phosphorylated p42/44 MAPK) in a time- and concentration-dependent manner. These results indicate that DADS can induce apoptosis in human nasopharyngeal carcinoma cells via, at least partly, S-phase block of the cell cycle, related to a rise in MAPK phosphorylation.     

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。     

Costunolide induces differentiation of human leukemia HL-60 cells

Costunolide has been reported to be a cytotoxic and chemopreventive agent. This work investigated the mechanism of the antiproliferative effect of costunolide and determined that it induced differentiation of the human leukemia cell line HL-60. Costunolide exhibited a potent antiproliferative activity against HL-60 cells. It was also found to be a potent inducer of differentiation in human leukemia derived HL-60 cells through the examination of differentiation markers, as assessed by the reduction of nitroblue-tetrazolium, the increase in esterase activities and phagocytic activity, morphology change and the expression of CD14 and CD66b surface antigens. These results, accompanied by a decline in the expression of c-myc protein, suggest that costunolide induces differentiation of human leukemia cells to granulocytes and monocytes/macrophages lineage.     

DADS抑制JAK1/STAT3信号通路诱导人白血病HL-60细胞分化

目的探讨JAKs/STATs信号转导通路在二烯丙基二硫(DADS)诱导人白血病HL60细胞分化中的变化及其调控机制。方法将HL60细胞与DADS或JAKs/STATs信号通路的激酶抑制剂AG490在体外共同培养,观察细胞形态变化,检测药物作用前后细胞NBT还原能力及细胞表面分化抗原CD11b的改变;用Westernblot检测JAKs,STATs各家族成员在DADS诱导HL60细胞分化中的改变;并用免疫细胞化学法检测核转录基因STATs,cmyc,cfos,cjun的表达变化。结果DADS和AG490均可诱导HL60细胞向成熟粒系分化,且DADS在1.25mg·L-1时诱导分化作用达峰值;Westernblot检测JAK1,STAT3的酪氨酸激酶发生了磷酸化改变;免疫细胞化学示STAT3与cmyc基因蛋白核内表达下降,cjun,cfos基因蛋白核内表达上升。结论JAK1,STAT3酪氨酸激酶的磷酸化抑制参与了DADS诱导HL60细胞分化的调控,其机制可能通过调控与HL60细胞增殖分化相关的基因表达,抑制细胞DNA合成,从而抑制细胞增殖,诱导分化。DADS的作用相当于JAK1/STAT3信号通路的阻断剂。     

Caspases家族成员促进DADAG诱导的人白血病HL—60细胞凋亡

目的:研究caspases家族成员在二乙酰二脱水卫矛醇(DADAG)诱导人白血病HL-60细胞凋亡中的作用.方法:MTT法观察DADAG的体外抗增殖作用;透射电镜、DNA梯形条带和流式细胞仪检测HL-60细胞凋亡;caspase-3检测试剂盒和Western blot法分析caspases家族成员.结果:DADAG明显抑制HL-60细胞增殖和诱导细胞凋亡.DADAG处理HL-60细胞24h后,caspase-3酶活性达峰值,同时聚腺苷二磷酸核糖聚合酶(PARP)、lamin B和DFF45蛋白开始出现断裂片段.Caspase-3抑制剂z-DEVD·fmk可部分逆转DADAG诱导的HL-60细胞凋亡,而caspases广谱抑制剂z-VAD·fmk可完全逆转此作用.结论:Caspases在DADAG诱导HL-60细胞凋亡中起重要作用,它们通过酶解底物PARP、DFF45和lamin B促进细胞凋亡.