黄芪对喉癌细胞增殖和凋亡的影响

目的体外观察黄芪对人喉癌细胞系Hep-2的抑制增殖和诱导凋亡作用并探讨其作用机制。方法用20、100、200μg/mL的黄芪作用于Hep-2细胞24 h,MTT法检测细胞抑制率,流式细胞仪检测细胞凋亡率,光学显微镜观察细胞形态学变化。结果 MTT结果显示,黄芪可显著抑制Hep-2细胞增殖且存在剂量依赖性;流式细胞仪检测发现,细胞凋亡率随着黄芪浓度增高逐渐升高,各实验组之间及其与对照组之间比较差异有统计学意义(P<0.05);用药后光镜观察细胞数量减少,荧光显微镜观察可见典型细胞凋亡。结论黄芪可调控喉癌细胞周期,形成G2/M期阻滞,通过抑制增殖和促进凋亡发挥其抗癌作用。     

柴胡皂苷B2(Saikosaponin B2)对四氯化碳损伤HepG2细胞的保护作用

目的初步探讨柴胡皂苷B2(SSB2)对CCl4肝细胞损伤的保护作用。方法用CCl4诱肝HepG2细胞损伤,设立正常对照组、CCl4损伤组和不同浓度SSB2保护组;利用四甲基偶氮唑蓝(MTT)法检测细胞活力以及流式细胞仪检测细胞周期和凋亡率的方法,观察SSB2对CCl4诱导的肝细胞损伤的保护作用。结果 MTT检测显示SSB2保护组细胞活性明显高于损伤组,以SSB2浓度为100μg/mL保护组细胞活性最高;流式细胞仪测定细胞凋亡率显示SSB2保护组(终浓度为100μg/mL)凋亡率明显低于CC14损伤组,细胞周期各期细胞分布比例无明显变化。结论 SSB2对CCl4诱导的HepG2细胞损伤具有保护作用。     

硝酸镓对卵巢癌细胞增殖与凋亡的作用

目的：探讨硝酸镓对卵巢癌HO-8910细胞诱导凋亡、抑制增殖的影响。方法通过对卵巢癌HO-8910细胞培养及干预（硝酸镓不同浓度及时间作用）,采用流式细胞仪技术对凋亡细胞进行测定;MTT技术对肿瘤细胞增殖进行测定。结果 MTT及流式细胞仪检测结果显示,增殖抑制率（80μg/ml）24 h为57.1%、48 h为59.4%、72 h为68.1%;凋亡率5μg/ml为（9.92±1.04）%、10μg/ml为（28.31±1.22）%、20μg/ml为（38.59±1.21）%、40μg/ml为（45.48±1.43）%、80μg/ml为（52.25±2.18）%,硝酸镓对HO-8910细胞作用呈剂量-时间±赖效应。结论硝酸镓对卵巢癌细胞作用明显,为肿瘤治疗及制备抗肿瘤药物提供新的途径。     

沙利度胺对人T淋巴细胞的免疫调节作用

目的 探讨沙利度胺(Thd)对健康人外周血T淋巴细胞的免疫调节作用.方法 健康人外周血T淋巴细胞经不同浓度Thd作用后,用MTT比色法检测其增殖情况,用流式细胞术检测其早期凋亡及CD28、CD152表达,用RT PCR检测其白细胞介素6(IL-6)、IL-10、肿瘤坏死因子α(TNF-α)mRNA表达水平.结果 在体外,与阴性对照组相比,500μg/ml Thd组可促进T淋巴细胞的增殖、早期凋亡,抑制T淋巴细胞CD28的表达.100μg/ml Thd组可促进T淋巴细胞CD152表达.各剂量组均能抑制IL-6、TNF-α mRNA表达,100μg/ml和500μg/ml Thd可增强IL-10 mRNA的表达.结论 Thd可以通过影响T淋巴细胞的增殖、早期凋亡以及CD152、CD28,IL-6、TNF-α、IL-10 mRNA的表达,对健康人T淋巴细胞具有免疫调节作用.     

吉非替尼(Iressa)对鼻咽癌CNE-Ⅰ细胞系抑制作用的研究

目的研究表皮生长因子受体酪氨酸激酶抑制剂吉非替尼(Iressa)在体外对鼻咽癌CNE-Ⅰ细胞系的抑制作用。方法MTT法检测Iressa对CNE-Ⅰ细胞的吸光度A550值并计算抑制率;培养瓶克隆形成率实验;流式细胞术检测CNE-Ⅰ细胞在10,30μg/mL的Iressa作用后24,48h细胞周期和凋亡率变化。结果MTT数据显示CNE-Ⅰ细胞的抑制率与Iressa浓度成正相关,随Iressa浓度升高,抑制作用越明显,经计算IC50=13.155μg/mL。培养瓶克隆形成率实验,对照组克隆形成率为33.2±1.8%,而10μg/mL Iressa组、30μg/mL Iressa组均为0,提示Iressa能明显抑制CNE-Ⅰ细胞增殖。流式细胞术分析显示Iressa处理48h使CNE-Ⅰ细胞凋亡率明显升高,且随浓度升高凋亡率升高越明显(P<0.001),细胞周期结果显示Iressa使细胞周期G0-G1期比例升高,S期和G2-M期比例下降,但除10μg/mL Iressa组G2-M期比例下降明显以外(P=0.042),其他各组间差异无显著性。结论Iressa在体外对鼻咽癌CNE-Ⅰ细胞有显著抑制增殖和促进凋亡作用。     

吉非替尼(Iressa)对鼻咽癌CNE-Ⅰ细胞系抑制作用的研究

目的研究表皮生长因子受体酪氨酸激酶抑制剂吉非替尼(Iressa)在体外对鼻咽癌CNE-Ⅰ细胞系的抑制作用。方法MTT法检测Iressa对CNE-Ⅰ细胞的吸光度A550值并计算抑制率;培养瓶克隆形成率实验;流式细胞术检测CNE-Ⅰ细胞在10,30μg/mL的Iressa作用后24,48h细胞周期和凋亡率变化。结果MTT数据显示CNE-Ⅰ细胞的抑制率与Iressa浓度成正相关,随Iressa浓度升高,抑制作用越明显,经计算IC₅₀=13.155μg/mL。培养瓶克隆形成率实验,对照组克隆形成率为33.2±1.8%,而10μg/mL Iressa组、30μg/mL Iressa组均为0,提示Iressa能明显抑制CNE-Ⅰ细胞增殖。流式细胞术分析显示Iressa处理48h使CNE-Ⅰ细胞凋亡率明显升高,且随浓度升高凋亡率升高越明显(P<0.001),细胞周期结果显示Iressa使细胞周期G0-G1期比例升高,S期和G2-M期比例下降,但除10μg/mL Iressa组G2-M期比例下降明显以外(P=0.042),其他各组间差异无显著性。结论Iressa在体外对鼻咽癌CNE-Ⅰ细胞有显著抑制增殖和促进凋亡作用。     

体外观察雷公藤内酯醇对乳腺癌细胞MCF-7增殖及凋亡的影响

目的:探讨雷公藤内酯醇(triptolide,TP)对人乳腺癌细胞MCF-7增殖及凋亡的影响。方法:MTT法检测不同条件下TP对MCF-7细胞的增殖抑制作用;倒置显微镜观察TP对MCF-7细胞形态学影响;流式细胞仪检测MCF-7细胞的凋亡情况;Caspase检测试剂盒测定Caspase-3,9的变化。结果:TP以剂量及时间依赖性方式明显抑制MCF-7细胞的增殖;TP作用MCF-7细胞后,细胞出现明显的形态学改变(形态不规则、脱落,细胞碎片等)。流式细胞仪检测5μg/mL的TP明显诱导MCF-7细胞凋亡;Caspase-3,9表达水平明显升高,和对照组相比差异有统计学意义(P<0.05)。结论:TP可能通过线粒体通路诱导MCF-7细胞凋亡而发挥其抑制作用。     

大蒜素对人横纹肌肉瘤RD细胞的作用及其机制

目的：探讨大蒜素（ allicin）对人横纹肌肉瘤RD细胞增殖和凋亡的作用及其相关机制。方法应用不同浓度大蒜素（5,10,20,30μg/ml）作用人横纹肌肉瘤RD细胞株24h,采用MTT比色法检测不同浓度大蒜素下RD细胞的增殖抑制率;流式细胞仪检测不同浓度大蒜素作用后RD细胞的凋亡率。结果 MTT比色法检测各实验组增殖抑制率分别为（25.82±1.75）%、（31.04±1.61）%、（36.32±1.61）%、（42.58±3.88）%;流式细胞仪检测对照组及各实验组凋亡率分别为（5.23±0.80）%、（7.07±0.83）%、（10.60±2.51）%、（15.70±1.79）%、（19.33±0.72）%。以上检测各组间比较差异均有统计学意义（ P<0.05）。结论大蒜素能有效抑制RD细胞的增殖,诱导其凋亡。     

螺内酯诱导人急性白血病细胞Jurkat凋亡的作用研究

目的研究螺内酯对人急性白血病细胞Jurkat体外增殖的抑制及诱导凋亡的作用。方法将终浓度分别是10、50及100μmol/L的螺内酯加入Jurkat细胞的培养体系中,24 h内每隔4 h通过MTT法分析Jurkat细胞的增殖抑制率,Annexin V/PI流式细胞术法分析Jurkat细胞的早期凋亡率。结果螺内酯与Jurkat细胞共同培养12 h后,螺内酯能显著增加对细胞的增殖抑制作用,并且与药物浓度成正相关,各浓度组的抑制率随着培养时间的延长而升高,与药物干预时间成正相关;螺内酯处理Jurkat细胞24 h,各浓度组诱导细胞凋亡率分别为:10μmol/L组(11.2±0.35)%、50μmol/L组(29.8±1.27)%及100μmol/L(56.5±1.41)%,各个浓度组诱导细胞凋亡率与药物浓度成正相关。结论螺内酯对人T淋巴细胞白血病Jurkat细胞具有抑制增殖和诱导凋亡的作用,提示该药可能具有潜在抑瘤作用。     

维生素E琥珀酸酯联合5-氟尿嘧啶抗人肝癌细胞增殖作用的研究

目的观察维生素E琥珀酸酯(VES)联合5-氟尿嘧啶(5-FU)抗人肝癌细胞SMMC-7721的增殖作用。方法体外培养的SMMC-7721细胞以VES联合5-FU分别作用24和48h,MTT法测定细胞增殖的抑制作用;流式细胞仪分析细胞周期、凋亡率以及Fas的表达。结果MTT检测显示,VES6μg/mL作用细胞24h后抑制率为(2.78±0.05)%,随药物浓度的增加和作用时间的延长,抑制率显著增加(P<0.01);相同条件下,VES与5-FU合用较两药单用抑制作用显著增加(P<0.01)。流式细胞仪分析显示,SMMC-7721细胞自然凋亡率为(0.86±0.20)%,VES24μg/mL,5-FU30μg/mL及两者合用作用细胞24h后凋亡率分别为(30.08±2.32)%,(18.23±1.58)%和(63.68±2.88)%(P<0.01),细胞表面Fas表达增加。结论VES联合5-FU通过阻滞细胞周期于G0/G1期、促进细胞表面Fas的表达协同抑制肝癌细胞增殖。     

Oral small-molecule tyrosine kinase inhibitor midostaurin (PKC412) inhibits growth and induces megakaryocytic differentiation in human leukemia cells

Midostaurin (PKC412), a small-molecule multiple tyrosine kinase inhibitor, has been shown to suppress the growth of various tumor cells. Since kinases inhibited by midostaurin are involved in megakaryocytic differentiation, we hypothesized that this novel target therapeutic might have a role in the treatment of human leukemia cells. Hence, we examined the effect of midostaurin on human erythroleukemia cells and evaluated potential mechanisms. Midostaurin inhibited the growth of both K562 and HEL cells in dose- and time-dependent manner. Morphological changes such as enlarged contours, multipolarity of mitotic spindles, and multinucleation of megakaryocytes were noted in both cell lines treated by midostaurin. A smaller population of apoptotic cells was also observed. DNA histogram revealed polyploid and hypoploid populations of midostaurin-treated cells. Midostaurin treatment enhanced the surface expression of the megakaryocytic marker CD61; in contrast, the erythroid marker glycophorin A expression was decreased. At optimal conditions for inducing megakaryocytic differentiation, midostaurin upregulated the expression and signaling of c-Mpl, a thrombopoietin receptor-encoding gene, in HEL cells. These results indicate that midostaurin can inhibit growth; induce megakaryocytic differentiation; and to a lesser extent, cause apoptosis in HEL cells. This effect might involve the expression and signaling of c-Mpl.     

Evi1基因特异性siRNA对HEL细胞生长周期的影响

目的研究Evi1基因特异性小干扰RNA(siRNA)对人红白细胞白血病(HEL)细胞增殖及生长周期的影响。方法将HEL细胞分为三组:Evi1基因siRNA组(A组)、siRNA-co对照组(B组)、细胞空白对照组(C组)。转染48 h后,台盼蓝染色实验检测细胞活力,流式细胞术检测细胞周期和凋亡率。结果与B、C组相比,A组HEL细胞活性下降[(97.13±1.58)%、(99.20±2.27)%vs.(26.05±2.49)%],细胞周期阻滞在G0/G1期[(39.61±1.32)%、(35.77±1.31)%vs.(79.07±1.43)%],S期细胞减少[(57.74±1.08)%(、57.36±1.38)%vs.(9.51±0.75)%],凋亡率增高[(9.78±1.14)%(、8.67±1.89)%vs.(49.94±1.75)%](P均<0.05);而B组和C组间各观察指标比较差异无统计学意义(P>0.05)。结论 Evi1基因特异性siRNA可抑制HEL细胞增殖,促进细胞凋亡。     

Dual mechanisms of action of the retinoid CD437: nuclear retinoic acid receptor-mediated suppression of squamous differentiation and receptor-independent induction of apoptosis in UMSCC22B human head and neck squamous cell carcinoma cells

The synthetic retinoid 6-[3-(adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid (CD437), which can bind to and activate the nuclear retinoic acid receptors beta and gamma (RARbeta/gamma), is a potent inducer of apoptosis in various cancer cell lines. However, this effect was reported to be independent of RARs. In this study, we compared and contrasted the potencies and mechanisms of action of CD437 and several other receptor-selective retinoids in induction of apoptosis and modulation of squamous differentiation in UMSCC22B human head and neck squamous cell carcinoma cell line. CD437 and the structurally related retinoid CD2325 exhibited almost equal potency in inducing apoptosis, whereas several other retinoids failed to induce apoptosis. The RAR-specific pan antagonist AGN193109 failed to suppress CD437-induced apoptosis, indicating that the induction of apoptosis by CD437 was RAR-independent. c-Fos expression was induced by CD437 and CD2325 that induced apoptosis in the cell line but not by other retinoids that failed to induce apoptosis, suggesting a role for c-Fos in CD437-induced apoptosis. At low concentration (0.01 microM), CD437 shared with several other receptor-selective retinoids the ability to suppress the mRNA levels of the squamous differentiation markers Spr1, involucrin, and cytokeratin 1. This effect of CD437 could be blocked by AGN193109. We conclude that CD437 can exert its effects in UMSCC22B human human head and neck squamous cell carcinoma cells by at least two mechanisms: RAR-mediated suppression of squamous differentiation and RAR-independent induction of apoptosis.     

消癌平注射液对白血病细胞NB4作用的初步研究

目的研究消癌平注射液对白血病细胞株NB4的效应并初步探讨其机制。方法采用台盼兰染色法检测消癌平注射液对白血病细胞株NB4细胞增殖和活力的影响。流式细胞术分析消癌平注射液对NB4细胞周期和细胞表面分化抗原CD11b表达的影响。NBT还原实验检测对NB4细胞分化的影响。免疫印迹方法检测对细胞周期和凋亡相关蛋白的影响。结果消癌平注射液能以时间和剂量依赖方式抑制白血病细胞NB4增殖。诱导NB4细胞发生G0/G1期阻滞,增加P21蛋白质的水平,但不改变P27的表达。对细胞凋亡相关蛋白质的PARP水平没有影响。消癌平注射液能增加NB4细胞表面抗原CD11b的表达,增加NBT阳性细胞数,诱导NB4细胞分化。结论消癌平注射液能通过诱导细胞周期阻滞和细胞分化抑制NB4细胞增殖。     

Effects of tumor necrosis factor alpha on replication of varicella-zoster virus.

Replication of varicella-zoster virus (VZV) and expression of VZV nuclear antigen are inhibited in human embryonic lung fibroblast (HEL) cells pretreated with recombinant tumor necrosis factor (TNF) alpha for 24 h. This antiviral activity is completely blocked by the addition of monoclonal antibodies against TNF. TNF acts synergistically with interferons alpha and gamma. When TNF is added to HEL cells after VZV adsorption, virus replication is still inhibited. When VZV-infected HEL cells are co-cultured with HEL cells which have been pretreated with TNF or grown in the presence of TNF, spread of VZV from VZV-infected HEL cells to uninfected cells is unaffected. No interferon is detected in the supernatants or cell lysates of HEL cells cultured with TNF and antibodies to alpha-, beta- and gamma-interferon have no effect on the antiviral action of TNF.     

曲古抑菌素对小细胞肺癌细胞系NCI-H446的诱导分化及凋亡作用

目的探讨曲古抑菌素对NCI-H446细胞的诱导分化及凋亡的作用机制。方法用巢蛋白、CD133、波形蛋白和CD44的抗体对NCI-H446细胞进行免疫荧光染色,鉴定该细胞的干细胞特性;用NF-200、βⅢ-Tubulin、微管相关蛋白(MAP2)和BM88、Ki67的抗体进行免疫荧光染色和免疫印迹测定,观察NCI-H446细胞经曲古抑菌素诱导后向神经细胞分化的成熟程度和细胞增殖指数的变化;用Caspase-3、Bax、Bcl-2表达水平的变化,评价曲古抑菌素对小细胞肺癌的抗癌效果。结果小细胞肺癌NCI-H446细胞高表达巢蛋白、CD133、波形蛋白和CD44;曲古抑菌素诱导NCI-H446细胞向神经细胞分化并激活Caspase-3,诱导细胞的Bax表达水平增高,Bcl-2表达水平降低,细胞的增殖指数明显降低,凋亡指数明显增高。结论曲古抑菌素可诱导小细胞肺癌NCI-H446细胞分化成熟,最终激活Caspase-3通路导致细胞凋亡。     

地西他滨联合丙戊酸钠对白血病细胞株HL-60促凋亡和分化的影响

目的探讨地西他滨（DCA）和丙戊酸钠（VPA）联用对白血病细胞株HL-60促凋亡和分化诱导的协同作用。方法白血病细胞株HL-60分别与以下药物终浓度组作用48 h:对照组,DCA单药A组(1.0μmol·L^-1),DCA单药B组(4.0μmol·L^-1),VPA单药组(2.0 mmol·L^-1),联合用药A组(IDCA 1.0μmol·L^-1+VPA 2.0 mmol·L^-1),联合用药B组(DCA 4.0μmol·L^-1+VPA2.0 mmol·L^-1)。应用Annexin V-FTTC/PI标记法检测早期凋亡率,流式细胞术检测CD34、CD117平均荧光强度（MFI）以及CD11b、CD14表达率。结果联合用药A、B组的凋亡率高于其各自的单药组,差异具有高度统计学意义（P<0.01）;联合用药A、B组的CD117和CD34 MFI低于其各自的单药组,差异具有高度统计学意义（P<0.01）;联合用药A组的CD11b和CD14表达率以及联合用药B组的CD11b表达率均低于其各自的单药组,差异具有高度统计学意义（P<0.01）。结论白血病细胞株HL-60中VPA能显著增强DCA的促凋亡分化作用。     

Valproic acid perturbs hematopoietic homeostasis by inhibition of erythroid differentiation and activation of the myelo-monocytic pathway

As a histone deacetylase inhibitor, valproic acid (VPA) is a candidate for anticancer therapy. Besides, VPA exhibits various mechanisms of action and its effects on the molecular basis of hematopoiesis remain unclear. To study the effects of VPA on the hematopoietic system, we performed microarray analysis using K562 cells treated with 1 mM VPA over a 72 h time course. The association between gene ontology (GO) terms and the lists of differentially expressed genes was tested using the Bioconductor package GOstats. Enrichment analysis for cellular differentiation pathways was performed based on manually curated gene lists. Results from microarray analysis were confirmed by studying cell differentiation features at the molecular and cellular levels using other hematopoietic cell lines as well as hematopoietic stem/progenitor CD34⁺ cells. Microarray analysis revealed 3440 modulated genes in the presence of VPA. Genes involved in the granulo-monocytic differentiation pathway were up-regulated while genes of the erythroid pathway were down-regulated. This was confirmed by analyzing erythrocytic and myeloid membrane markers and lineage-related gene expression in HEL, MEG01, HL60 as well as CD34⁺ cells. Moreover, GATA-1 and its co-factors (FOG1, SP1) were down-regulated, while myelopoiesis activator PU.1 was up-regulated, in agreement with an inhibition of erythropoiesis. Our functional profiling and cell phenotyping approach demonstrates that VPA is able to alter hematopoietic homeostasis by modifying the cell population balance in the myeloid compartment. This may lead to a potential failure of erythropoiesis in patients with cancer or chronic inflammatory diseases having a well-described propensity to anemia.