DNA引物酶抑制剂碘化-3,3′-二乙基-9-甲基-硫杂羰花青诱导人白血病HL-60细胞凋亡

目的研究DNA引物酶抑制剂碘化-3,3′-二乙基-9-甲基-硫杂羰花青(DMTCCI)诱导人粒细胞性白血病HL-60细胞凋亡并探索其机制。方法分别采用不同浓度的DMTCCI处理培养于RPMI-1640培养基的HL-60细胞。采用MTT法检测DMTCCI对HL-60细胞的生长抑制作用。采用流式细胞仪和DNA琼脂糖凝胶电泳方法检测细胞凋亡。采用蛋白免疫印迹(Western blotting)法观察凋亡相关蛋白survivin， Bcl-xL， Bad， Bax， Bcl-2， caspase-9， caspase-3， caspase-6， PARP， DFF45和lamin B的表达。采用ApoAlert Caspase-3分析试剂盒检测caspase-3的活性。结果DMTCCI具有抑制人白血病HL-60细胞增殖的作用，其IC₅₀值为0.24 μmol·L^-1。流式细胞仪和DNA琼脂糖凝胶电泳结果显示，DMTCCI可诱导HL-60细胞凋亡。在经DMTCCI处理的HL-60细胞中，survivin和Bcl-xL蛋白的表达水平下调，Bad和Bax蛋白的表达水平上调，Bcl-2蛋白的表达水平无变化，caspase-9，caspase-3，caspase-6，PARP，DFF45和lamin B被分别裂解，产生相应裂解产物。在HL-60细胞中，caspase-3的活性在1 μmol·L^-1 DMTCCI处理3 h时明显升高，在处理12 h时达到最高峰。结论DMTCCI可抑制人白血病HL-60细胞的增殖并诱导其发生细胞凋亡。Bcl-2家族蛋白、survivin和caspases家族蛋白可能参与了上述诱导HL-60细胞凋亡的过程。     

DNA引物酶抑制剂碘化-3,3′-二乙基-9-甲基-硫杂羰花青诱导人白血病HL-60细胞凋亡(英文)

目的研究DNA引物酶抑制剂碘化-3,3′-二乙基-9-甲基-硫杂羰花青(DMTCC I)诱导人粒细胞性白血病HL-60细胞凋亡并探索其机制。方法分别采用不同浓度的DMTCC I处理培养于RPM I-1640培养基的HL-60细胞。采用MTT法检测DMTCC I对HL-60细胞的生长抑制作用。采用流式细胞仪和DNA琼脂糖凝胶电泳方法检测细胞凋亡。采用蛋白免疫印迹(W estern b lotting)法观察凋亡相关蛋白survivin,Bc l-xL,Bad,Bax,Bc l-2,caspase-9,caspase-3,caspase-6,PARP,DFF45和lam in B的表达。采用ApoA lert Caspase-3分析试剂盒检测caspase-3的活性。结果DMTCC I具有抑制人白血病HL-60细胞增殖的作用,其IC50值为0.24μmol.L-1。流式细胞仪和DNA琼脂糖凝胶电泳结果显示,DMTCC I可诱导HL-60细胞凋亡。在经DMTCC I处理的HL-60细胞中,survivin和Bc l-xL蛋白的表达水平下调,Bad和Bax蛋白的表达水平上调,Bc l-2蛋白的表达水平无变化,caspase-9,caspase-3,caspase-6,PARP,DFF45和lamin B被分别裂解,产生相应裂解产物。在HL-60细胞中,caspase-3的活性在1μmol.L-1DMTCCI处理3 h时明显升高,在处理12 h时达到最高峰。结论DMTCCI可抑制人白血病HL-60细胞的增殖并诱导其发生细胞凋亡。Bcl-2家族蛋白、survivin和caspases家族蛋白可能参与了上述诱导HL-60细胞凋亡的过程。     

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.     

二乙酰二脱水卫矛醇诱导白血病HL-60细胞凋亡及其机理

目的研究二乙酰二脱水卫矛醇(DADAG)诱导人白血病HL-60细胞凋亡及其机理。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;Western blotting法和caspase-3检测试剂盒分析DADAG诱导HL-60细胞凋亡与Bcl-2家族成员和caspase-3的关系。结果DADAG明显抑制HL-60细胞增殖和诱导细胞发生凋亡。8 μg·mL^-1 DADAG处理HL-60细胞不同时间后,Bcl-X_L蛋白水平呈时间依赖性地下降,而Bad蛋白水平上调。DADAG处理HL-60细胞24 h后,caspase-3酶活性达峰值。Caspase-3抑制剂z-DEVD.fmk可部分逆转DADAG诱导HL-60细胞凋亡的作用,而caspases广谱抑制剂z-VAD.fmk可完全逆转此作用。结论DADAG诱导HL-60细胞凋亡依赖caspase-3途径的激活,而caspase-3的激活可能与Bcl-2家族成员密切相关。     

二氢青蒿素诱导HL-60细胞凋亡

目的探讨二氢青蒿素对人早幼粒白血病细胞(HL-60)的治疗作用。方法采用台盼蓝染色法和MTT法测定二氢青蒿素对HL-60细胞存活的影响,吖啶橙/溴化乙啶(AO/EB)荧光双染色、DNA凝胶电泳、流式细胞术检测细胞凋亡,Western印迹分析凋亡相关蛋白的表达。结果二氢青蒿素可抑制HL-60细胞存活,诱导HL-60细胞凋亡,同时降低HL-60细胞Bcl-2蛋白的表达,增加Bax蛋白和凋亡执行蛋白半胱氨酸天冬氨酸蛋白酶(caspase)-3的表达,并呈浓度依赖性。结论二氢青蒿素可诱导HL-60细胞凋亡,其机制可能是通过对线粒体凋亡通路中Bcl-2,Bax和caspase-3蛋白表达的调控而发挥作用。     

番荔枝内酯诱导白血病细胞凋亡的机理

目的　研究番荔枝内酯(squamocin)诱导白血病细胞凋亡的机理。方法　DNA凝胶电泳法、荧光染色等检测细胞凋亡。试剂盒检测caspase-3的活性。Westernblot法检测PARP和caspase-3的剪切片断和磷酸化的SAPK/JNK量的变化。结果　Squamocin处理HL-60细胞后,导致染色质浓缩、片断化,DNA梯形条带出现,完整PARP的116ku条带逐渐减少,而85ku的片断逐渐增加,caspase-3酶的活性也增加。Caspase-3的特异性抑制剂DEVD-CHO预处理HL-60细胞,可阻止squamocin诱导的DNA片断化、PARP的剪切和细胞死亡。Squamocin激活HL-60细胞中SAPK/JNK。结论　Squamocin诱导HL-60细胞凋亡依赖caspase-3途径的激活,squamocin激活caspase-3可能与SAPK/JNK的激活相关     

去甲斑蝥素诱导人黑色素瘤A375-S2细胞凋亡

目的:研究去甲斑蝥素(NCTD)诱导人黑色素瘤A375-S2细胞凋亡的机制.方法:采用MTT法、形态学观察、DNA凝胶电泳及Western blot检测法.结果:去甲斑蝥素可诱导A375-S2细胞发生凋亡.半胱氨酸天冬氨酸酶(caspase)-3,-9抑制剂可以部分的抑制NCTD诱导的细胞死亡.细胞凋亡时caspase-3,8,-9酶活力升高,caspase-3底物-caspase-3激活的DNA酶抑制物(ICAD)蛋白表达下降,同时Bcl-2/Bax、Bcl-xL/Bax蛋白表达比率明显降低.结论:去甲斑蝥素通过激活caspase和Bcl-2家族诱导A375-S2细胞凋亡.     

熊果酸对脑胶质瘤细胞株U251凋亡及survivin和caspase-3表达的影响

目的 观察熊果酸（ursolic acid, UA）诱导脑胶质瘤细胞株U251凋亡的机制. 方法 培养人脑胶质瘤细胞株U251,以Hoechst 33258染色法及琼脂糖凝胶电泳测定UA诱导脑胶质瘤细胞株U251凋亡的作用. 免疫印记法及荧光双标法测定survivin和caspase-3表达,荧光法测定caspase-3活性. 结果 Hoechst 33258染色法和琼脂糖凝胶电泳显示UA明显诱导U251细胞凋亡,细胞核DNA呈梯状降解. 免疫沉淀法以及荧光双标法均显示UA可抑制survivin表达;同时UA上调caspase-3表达. 结论 UA诱导U251细胞凋亡与其抑制survivin表达,同时上调caspase-3表达有关.     

大萼香茶菜甲素通过线粒体途径诱导HL-60白血病细胞凋亡

目的：观察大萼香茶菜甲素（macrocalyxinA,MA）体外诱导HL-60细胞凋亡,并探讨其作用机制。方法：不同浓度的MA与HL-60细胞进行培养,采用MTT比色法观察其对HL-60细胞增殖的抑制作用;细胞形态学、DNA含量、DNA梯度电泳及细胞周期分析、Annexin—V／PI双标记和Hoechst 33258荧光染色等分析其促细胞凋亡效应;流式细胞术和RT-PCR分别检测Bcl-2、Bax、P53、Fas、线粒体膜蛋白（Ap02．7）、线粒体跨膜电位（AWm）与Bcl-2、Bax、P53、caspase-3 mRNA变化水平,研究其促凋亡机制。结果：MA呈现作用时间和剂量依赖性地抑制HL-60细胞增殖和活力;HL-60细胞经MA作用后,Wright—Giemsa染色和Hoechst荧光染色后细胞出现典型的凋亡小体,细胞阻滞于G0／G1期,DNA片段化,亚二倍体明显增高,Annexin—V／PI标记升高;MA诱导HL-60细胞凋亡过程中,Bcl-2、Fas、P53表达无明显变化,Bax、线粒体膜蛋白（Apo2．7）、caspase-3表达显著增加,Bax／Bcl-2比值升高,龇下降。结论：MA能抑制HL-60细胞增殖和细胞活力、诱导细胞凋亡,其机制通过上调Bax基因和Bax／Bcl-2比值,使线粒体膜电位下降、膜通透性增高,最终使caspase-3激活而促进凋亡。     

异十三基二乙胺诱导HL-60细胞凋亡及其作用机制

目的探讨异十三基二乙胺(D-108)体外诱导人早幼粒白血病HL-60细胞凋亡及其作用机制。方法应用生长曲线法检测D-108对HL-60细胞生长抑制作用;相差显微镜观察药物对HL-60细胞促凋亡形态学变化;流式细胞仪分析细胞周期改变;DNA琼脂糖凝胶电泳法检测凋亡"梯状"DNA;比色法测定caspase-3,8,9酶活性:结果D-108显著抑制HL-60细胞生长,作用呈浓度及时间依赖性。HL-60细胞经D-108处理后,呈典型凋亡形态学改变,出现亚二倍体细胞峰及DNA梯状条带。caspase-3,9酶活性显著升高(P<0.01),而对caspase-8酶活性无明显影响。结论D-108在体外能有效诱导HL-60细胞凋亡,其作用机制可能通过线粒体凋亡途径。     

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.     

牛蒡子苷元诱导人白血病细胞凋亡的作用及机制

牛蒡子来源于菊科两年生草本植物牛蒡子属牛蒡(Aictium lappa L.)的干燥成熟果实,为常用中药.有疏风散热、解毒透疹、利咽消肿等功效,现代药理学研究表明牛蒡子有抗病毒[1,5]、抗炎[2]、抗癌[3,4]、抑制热休克反应[6]等广泛的药理活性.其主要活性成分是牛蒡子苷元(arctigenin,ARG)[1].     

Bcl-xL is a dominant antiapoptotic protein that inhibits homoharringtonine-induced apoptosis in leukemia cells

Homoharringtonine (HHT) has been reported to be effective in a portion of patients with acute myeloid leukemia (AML) or chronic myeloid leukemia (CML). To investigate its mechanism of action, cell growth inhibition and cytotoxicity of HHT were investigated in three AML cell lines, HL-60, NB4, and U937, and in three CML cell lines, K562, KU812, and KCL22. AML cells were more sensitive than CML cells to HHT-induced cytotoxicity. Using HL-60 cells, it was revealed that HHT decreased the levels of myeloid cell leukemia 1 (Mcl-1), X-linked inhibitor of apoptosis protein (XIAP), survivin, and B-cell lymphoma 2 (Bcl-2)-homology domain 3 (BH3)-only proteins as well as the mitochondrial membrane potential. The levels of Bcl-2, Bcl-2-associated X protein (Bax), and Bcl-2 homologous antagonist/killer (Bak) proteins in HL-60 cells were not changed after HHT treatment. U937, K562, KU812, and KCL22 cells expressed B-cell lymphoma-extra large (Bcl-xL) and were less responsive to HHT-induced apoptosis than HL-60 cells. Silencing Mcl-1 or Bcl-xL, but not XIAP or survivin, enhanced HHT-induced apoptosis in U937 cells. The levels of HHT-induced apoptosis in K562, KCL22, and KU812 cells were inversely correlated with the levels of Bcl-xL but not those of Bcl-2 or Mcl-1. K562 cells expressing high levels of Bcl-xL but no Bcl-2 were less responsive to HHT-induced apoptosis than KCL22 cells that expressed lower levels of Bcl-xL and higher levels of Bcl-2 protein. In K562 cells, knockdown of Bcl-xL, but not of Mcl-1, enhanced HHT-induced apoptosis. Transfection of Bcl-xL into KCL22 cells attenuated HHT-induced apoptosis. These data suggest that Bcl-xL plays a more important role than Bcl-2 and Mcl-1 in protecting against HHT-induced apoptosis.     

Effects of a naphthoquinone analog on tumor growth and apoptosis induction

Vitamin K-related analogs induce growth inhibition in various cancer cell lines. A naphthoquinone analog, termed 2,3-dichloro-5, 8-dihydroxy-1,4-naphthoquinone (DDN), induces apoptosis in human promyeloid leukemic HL-60 cells, and shows antitumor activity in vivo. Following treatment with DDN, evidence of apoptosis, including DNA fragmentation and cleavage of poly ADP ribose polymerase (PARP), was observed. DDN induced an upregulation of proapoptotic Bax protein, and Bid cleavage. Antiapoptotic Bcl-2 protein levels were not changed by DDN, but the expression of Bcl-xL was decreased. In addition, DDN reduced the mass of solid tumor in the Sarcoma 180 tumor-bearing mouse model. These results indicate that DDN exerts antitumor activity, which appears to be related to the induction of apoptosis by regulating Bcl-2 family proteins.     

The cytotoxicity of eutigosides fromEurya emarginata against HL-60 promyelocytic leukemia cells

Two phenolic glucosides, eutigoside B and eutigoside C were isolated from the fresh leaves of Eurya emarginata. These two phenolic glucosides exerted a significant inhibitory effect on the growth of HL-60 promyelocytic leukemia cells. Furthermore, when the HL-60 cells were treated with eutigoside C, several apoptotic characteristics such as DNA fragmentation, morphologic changes, and increase of the population of sub-G1 hypodiploid cells were observed. In order to understand the mechanism of apoptosis induction by eutigoside C, we examined the changes of Bcl-2 and Bax expression levels. The eutigoside C reduced Bcl-2 protein and mRNA levels, but slightly increased Bax protein and mRNA levels in a time-dependent manner. When we examined the activation of caspase-3, an effector of apoptosis, the eutigoside C increased the expression of active form (19-kDa) of caspase-3 and the increase of their activities was demonstrated by the cleavage of poly (ADP-ribose) polymerase, a substrate of caspase-3, to 85-kDa. The results suggest that the inhibitory effect of eutigoside C from E. emarginata on the growth of HL-60 appears to arise from the induction of apoptosis via the down-regulation of Bcl-2 and the activation of caspase.