丹参酮ⅡA与全反式维甲酸协同诱导急性早幼粒细胞白血病细胞株的分化和凋亡

目的研究丹参酮ⅡA(Tanshinone ⅡA, TanⅡA)联合全反式维甲酸(ATRA)对早幼粒细胞白血病细胞(NB4)诱导分化及凋亡的协同效应.方法0.5 μg/ml TanⅡA分别联合0.5 μg/ml、0.25 μg/ml和0.125 μg/ml ATRA作用NB4细胞5 d,观察NB4细胞形态学变化,检测硝基蓝四氮唑(NBT)还原能力;流式细胞术检测CD11b,CD33的表达,并进行细胞周期和细胞凋亡分析.结果TanⅡA联合ATRA可协同诱导NB4细胞分化和凋亡.联合作用组第5 d细胞生长抑制率均超过80%;90%左右的NB4细胞分化,其中杆状和分叶核细胞比例超过65%;NBT还原能力显著升高.流式细胞术分析显示CD11b表达升高,CD33表达降低;G0/G1期细胞增加,有明显的细胞凋亡,与TanⅡA或ATRA单独作用组相比均有显著性差异(P<0.01).TanⅡA与ATRA(0.125 μg/ml～0.5 μg/ml)联合对NB4细胞的影响没有明显的浓度依赖倾向.结论TanⅡA联合ATRA对NB4细胞诱导分化和凋亡有协同作用;在ATRA一定浓度范围内,这种协同作用无ATRA剂量依赖性.     

冬凌草甲素对急性白血病原代细胞的增殖抑制作用

目的研究冬凌草甲素对急性白血病原代细胞的增殖抑制作用。方法以人急性早幼粒细胞白血病(APL)患者的骨髓原代细胞为研究对象,以不同浓度的冬凌草甲素作用于体外培养的白血病细胞,MTT法检测细胞生长抑制率,应用流式细胞仪、台盼蓝染色及Hoechst33258荧光染色法观察细胞凋亡。结果冬凌草甲素体外可明显地抑制白血病原代细胞的增殖,并能诱导细胞发生凋亡,而且呈现出明显的量-效与时-效关系。结论冬凌草甲素能有效地抑制APL细胞的生长及诱导细胞发生凋亡,是一种潜在的抗白血病药物。     

A study on apoptosis of nasopharyngeal carcinoma cell line induced by Tanshinone II A and its molecular mechanism]

OBJECTIVE: To assess effect of Tanshinone II A on the apoptosis of human nasopharyngeal carcinoma (NPC) cell line CNE-1 and inquire into the mechanism there in involved. METHODS: The CNE-1 cells cultured in vitro were treated with 0.5 microgram/ml Tanshinone II A for 4 days. The morphology of the cells was observed by microscopy. The cell growth and proliferation were measured by cell counting. The DNA break was examined by gel electrophoresis. The cell cycle, apoptosis index (AI) and the expression of apoptosis associated gene were analysed by flow cytometry. RESULTS: 0.5 microgram/ml Tanshinone II A could induce the apoptosis of CNE-1 cells. In the treatment group, the morphologic characteristics of apoptotic cells were observed, the DNA of cells presented "ladder" break, the cell growth and proliferation were inhibited obviously, and the AI was 16.9% (the AI of control group was 6.4%). The cells were arrested in G0/G1 phase and cellular DNA synthesis was inhibited. The expressions of apoptosis associated gene fas, bax, p53 and p21 were up-regulated; the expression of bcl-2 was down-regulated. CONCLUSION: Tanshinone II A can induce apoptosis of human nasopharyngeal carcinoma cells. Its molecular mechanism may relate to modulation of the apoptosis associated gene expression.     

水溶性壳寡糖对人白血病细胞诱导分化的影响

目的:研究水溶性壳寡糖(WSC)抑制人白血病细胞(HL-60)增殖活性的作用。方法:将WSC与HL-60细胞共培养,四甲基偶氮唑盐(MTT)比色法检测WSC对HL-60细胞增殖的抑制作用;氮蓝四唑(NBT)法检测WSC对HL-60细胞分化促进作用;流式细胞仪定量检测细胞凋亡。结果:250～1000mg/L的WSC能抑制HL-60细胞增殖,并促进HL-60细胞分化,WSC作用48h后流式细胞仪分析图上出现凋亡峰。结论:WSC对诱导人白血病HL-60细胞凋亡及促进分化有一定作用     

丹参酮与三氧化二砷协同诱导NB4细胞株分化的体外研究

目的评估丹参酮ⅡA（Tanshinone ⅡA,Tan ⅡA）与三氧化二砷（As2O3）联合应用对急性早幼粒细胞白血病（NB4）细胞增殖、分化的效应。方法以不同浓度的As2O3、Tan ⅡA单独及联合作用于NB。细胞,观察细胞的生长状况、形态学改变,用流式细胞术（FCM）检测细胞膜表面CD11b,分析NB4细胞的增殖、分化情况。结果TanⅡA、As2O3单独均能抑制NB4细胞生长,以1．0μmol·L^-1 As2O3作用更显著,两药联合应用可增强对NB。细胞增殖的抑制作用;经Tan ⅡA处理的NB4细胞,其形态改变更多显示一定程度的成熟粒细胞的特征;NB4细胞表面CD11b检测显示Tan ⅡA的诱导分化能力更强,以0．5μg·mL^-1组为著,联合应用As2O3可拮抗其诱导分化能力,相反TanⅡA则可促进As2O3对NB4细胞的诱导分化作用。结论两药对NB4细胞均有增殖抑制作用,且具有协同效应;两药对NB4细胞诱导分化作用表现为：As2O3可以拮抗Tan ⅡA对NB4细胞的诱导分化能力,而Tan ⅡA则促进As2O3对NB4细胞的诱导分化作用。     

鼠尾草酸联合1,25-二羟基维生素D3对NB4细胞的影响

目的 研究鼠尾草酸(carnosic acid,CA)与1,25-二羟基维生素D3[1,25( OH)2D3]联合应用对NB4细胞生长及凋亡、分化的诱导作用.方法 通过四氮唑蓝(MTT)法观察细胞的生长,光学显微镜观察细胞形态,应用流式细胞仪测定细胞周期、凋亡率及CD14的表达,观察联合应用鼠尾草酸与低浓度1,25(O...     

5-杂氮-2′-脱氧胞苷抗白

目的:研究甲基转移酶抑制荆5-杂氮-2′-脱氧胞苷(5-aza-2dC)对人急性髓系白血病细胞株HL-60生长、分化、凋亡的影响,初步探讨其抗白血病作用的可能机制.方法:不同浓度和时间的5-aza-2dC处理HL-60细胞后,采用MTT比色试验检测5-aza-2dC对HL-60细胞生长的影响;采用流式细胞术检测5-aza-2dC对HL-60细胞周期及分化的影响;采用Hochest33342染色和流式细胞术检测5-aza-2dC时HL-60细胞凋亡的影响;采用RT-PCR法检测药物处理对S100A8和S100A9基因mRNA表达水平的影响.结果:(1)5-aza-2dC呈剂量和时间依赖性地抑制HL-60细胞的生长,并使HL-60细胞周期阻滞于G2/M期;(2)5-aza-2dC处理使HL-60细胞的髓系分化抗原CD11b的表达增强,在低浓度(0.5 μmol/L)时其促分化作用最明显;(3)5-aza-2dC呈剂量和时间依赖性地诱导HL-60细胞凋亡,在高浓度(5.0μmol/L)时诱导凋亡的作用最明显;(4)5-aza-2dC能明显上调S100A8和S100A9基因mRNA的表达.结论:5-aza-2dC能抑制HL-60细胞生长,阻滞HL-60细胞于G2/M期,促进HL-60细胞分化和诱导其凋亡,并能上调S100A8和S100A9基因的表达,这些作用可能是5-aza-2dC抗急性髓系白血病的重要机制.     

In vitro study on arsenic sulfide(realgar)-induced apoptosis of retinoic acid susceptible or resistant acute promyelocytic leukemia cell lines

Objective: To further understand the possible mechanisms of arsenic sulfide (realgar) in the treatment of acute promyelocytic leukemia (APL). Methods: All-trans retinoic acid (ATRA)-susceptible APL cell line (NB4 cells) and ATRA-resistant APL cell line (MR2 subclone) were used as models in vitro. At various times after incubated with various concentrations of realgar, NB4 and MR2 cells were observed by cell viability , cell proliferation and cell morphology; cell cycle and the expression of Annexin V were assayed by flow cytometry. Results: Cell viability and proliferation of NB4 and MR2 cells were inhibited after the treatment, to some extent, in a dose and time dependent manner. 177 - 711g/L of realgar treated NB4 and MR2 cell presented morphologically some features of apoptotic cells such as intact cell membrane, chromatin condensation and nuclear fragmentation, apoptosis body could be found by electron microscopy as well. Sub-Gl cells and cell cycle arrest were observed by flow cytometry. The proport     

氨基葡萄糖硫酸盐抑制白血病细胞HL60增殖并诱导其分化

目的:研究氨基葡萄糖硫酸盐(GS)对白血病细胞HL60的增殖和分化的影响. 方法:采用台盼蓝活细胞计数法检测不同浓度的GS对HL60细胞增殖的影响;细胞化学染色法观察药物作用后细胞形态的变化;流式细胞仪检测药物对HL60细胞周期的影响及细胞表面分化抗原CD11b,CD14,CD13和CD33表达的影响. 结果:GS能明显抑制HL60的增殖;经5 mmol/L GS处理5 d后,HL60细胞体积缩小,核浆比例降低,核仁减少甚至消失,核染色质趋向致密,核形态扭曲折叠或分叶;5 mmol/L GS处理48 h后, G1期细胞由37.8%增加至47.9%,S期由46.8%减低至40.6%,G2期峰轻度减低, 未发现亚二倍体峰. 经1 mmol/L,5 mmol/L GS分别处理120 h后,CD11b和CD14的表达分别由2.3%,0.5%升高至23.0%,11.9%和15.8%, 4.3%,CD33及CD13的表达未见明显变化. 结论:GS能抑制白血病细胞HL60的增殖,并诱导其向成熟单核、粒细胞分化.     

丹参酮ⅡA诱导NB4细胞凋亡时Caspase3活性变化

目的：研究Caspase3在丹参酮ⅡA(TanⅡA)诱导NB4细胞凋亡过程中的变化及与凋亡的关系。方法：通过细胞形态学、DNA含量分析及DNA片段比率，证实TanⅡA是否可诱导NB4细胞凋亡，用荧光分光光度仪检测Caspase3活性及N-乙酰-天冬氨酸-谷氨酸-颉氨酸-天冬氨酸-7氨基-4甲基-香豆素(AC-DEVD-AMC)行Caspase3抑制试验探讨细胞凋亡与Caspase3间的关系。结果：TanⅡA可诱导NB4细胞凋亡，并伴有Caspase3活性增高。AC-DEVD-乙醛(AC-DEVD-CHO)可抑制TanⅡA诱导NB4细胞凋亡。结论。TanⅡA诱导NB4细胞凋亡可通过激活Caspase3而实现。