一定浓度下三氧化二砷对T细胞淋巴瘤Hut-78细胞株增殖的影响

背景：近年来有将三氧化二砷试用于T细胞肿瘤的有效报道,但未见其治疗机制的相关研究。目的：检测三氧化二砷对人皮肤T细胞淋巴瘤Hut-78细胞的抑制增殖、诱导凋亡及细胞周期的影响。方法：分别采用MTT、PI单标记流式细胞术和TUNEL法检测2,5,10μmol/L的三氧化二砷干预24,48,72h,对人皮肤T细胞淋巴瘤Hut-78细胞株抑制增殖、诱导凋亡和细胞周期的影响,及其凋亡相关基因bcl-2及血管内皮细胞生长因子基因表达的变化。结果与结论：在一定的浓度范围内,三氧化二砷对Hut-78细胞存在增殖抑制作用,其增殖抑制作用在一定程度上可能与下调血管内皮细胞生长因子基因的表达有关,同时还存在一定的细胞毒作用;三氧化二砷可诱导Hut-78细胞发生凋亡,凋亡机制主要发生在G2～M期,其凋亡作用可能与下调bcl-2基因表达有关。三氧化二砷对Hut-78细胞的增殖抑制及诱导凋亡作用呈时间剂量依赖性。     

三氧化二砷体外诱导宫颈癌细胞株凋亡及其机制的研究

目的:研究三氧化二砷体外能否诱导宫颈癌细胞株(HeLa细胞)发生凋亡及其相关机制。方法:采用透射电镜、DNA Ladder、TUNEL的方法进行凋亡检测,用ECL Westernblot的方法检测凋亡过程中相关基因表达的变化。结果:三氧化二砷体外作用于HeLa细胞后,透射电镜观察到了凋亡小体;DNA Ladder法检测到了凋亡时DNA降解形成的梯带;TUNEL法也检测到HeLa细胞有DNA的断裂。在凋亡过程中,凋亡相关基因bcl-2的表达下调,而p53基因的表达没有明显改变。结论:三氧化二砷体外能诱导HeLa细胞发生凋亡,其机制与bcl-2基因的表达下调有关。     

藏红花素对HL-60细胞增殖抑制和诱导凋亡作用及其机制

本研究探讨藏红花素(crocin)对人急性白血病细胞HL-60增殖抑制的影响及其促凋亡机制。测定HL-60细胞在不同浓度crocin作用后的生长曲线,用荧光显微镜观察细胞形态学变化,MTT法检测细胞的增殖情况,流式细胞术检测细胞周期分布,RT-PCR检测bcl-2、bax基因的表达。结果表明,不同浓度crocin作用后HL-60细胞生长明显受抑制,并呈现出剂量时间依赖关系。在0-5mg/ml范围内HL-60细胞凋亡率逐渐升高。当crocin浓度高于5mg/ml时,HL-60细胞凋亡率并没有升高,反而下降,表现为细胞坏死。流式细胞术检测结果显示,细胞周期被阻滞于G0/G1,且在5mg/ml时阻滞作用最明显。RT-PCR结果显示,bcl-2基因表达明显下调,bax基因表达上调。结论:crocin可诱导HL-60细胞凋亡,并对细胞周期起阻滞作用,其作用机制可能与bcl-2基因表达抑制及bax基因表达激活有关。     

银耳多糖诱导肝癌HepG-2细胞凋亡的研究

目的:研究银耳多糖(Tremdla Polysacchadd,TP)对肝癌HepG-2细胞体外增殖的影响.方法:采用MTT法检测TP对HepG-2细胞的抑制作用,改良苯酚品红染色观察细胞形态的改变,梯状DNA电泳检测细胞凋亡,半定量RT-PCR检测bcl-2和survivin的表达.结果:TP对HepG-2细胞具有明显的抑制作用且呈剂量-时间依赖性;细胞有明显染色质凝集、凋亡小体等现象;电泳出现梯状DNA现象;抗凋亡基因bcl-2和survivin表达下调.结论:TP可诱导HepG-2细胞凋亡,而抗凋亡基因bcl-2和survivin下调可能是其诱导凋亡的机制之一.     

三氧化二砷对K562/ADM耐药细胞凋亡抑制的逆转作用

目的:研究三氧化二砷(As2O3)诱导白血病K562/ADM耐药细胞凋亡的分子机制。方法:采用MTT比色法检测K562/ADM耐药细胞增殖活性,细胞形态学和annexinV /PI双染色检测细胞凋亡,RT-PCR检测mdr1、bcl-2和caspase-3基因mRNA的表达水平,流式细胞法(FCM)测定P-糖蛋白(P -glycoprotein,P-gp)和bcl-2蛋白表达及caspase-3活性。结果:As2O3显著抑制K562/ADM耐药细胞的增殖;经 As2O3处理后细胞形态上出现典型的凋亡改变,annexinV /PI双染显示凋亡细胞明显增加;mdr1mRNA表达和P-gp合成明显降低,凋亡抑制基因bcl-2mRNA及其蛋白bcl-2表达下调, caspase-3mRNA表达和caspase-3活性显著增强。结论:As2O3诱导K562/ADM耐药细胞凋亡,其主要机制可能为As2O3抑制 mdr1和bcl-2基因表达,逆转耐药白血病细胞因bcl-2和P-gp高表达所介导的凋亡抑制。     

白藜芦醇诱导食管癌Eca109细胞凋亡的实验研究

目的探讨白藜芦醇（Res）对食管癌Eca109细胞体外增殖的影响，进而观察Res对Eca109细胞凋亡的影响。方法MTT法测定肿瘤细胞生长抑制率；流式细胞术（FCM）分析细胞周期，检测细胞凋亡；RT-PCR法检测细胞bcl-2和bax基因的表达。结果Res呈浓度、时间依赖性抑制Eca109细胞的生长与增殖，可以诱导Eca109细胞凋亡，细胞周期呈明显的G0／G1期阻滞现象。Eca109细胞的bcl-2基因表达降低，而bax基因表达增强。结论Res能通过诱导Eca109细胞凋亡而抑制其生长与增殖，其机制可能与抑制bcl-2，促进bax的表达有关。     

三氧化二砷诱导淋巴瘤Raji细胞凋亡与细胞周期阻滞

本研究探讨研究三氧化二砷（As2O3）对人淋巴瘤Raji细胞株凋亡诱导的作用特点及其机制。用MTT比色法观察不同浓度As2O3对Raji细胞的增殖抑制效应，电子显微镜观察不同浓度As2O3作用不同时间后细胞的凋亡形态，DNA-ladder琼脂糖凝胶电泳观察凋亡条带，流式细胞术检测Raji细胞凋亡、细胞周期分布。结果表明：1—8μmol／LAs2O3能明显抑制Raji细胞的活力，并存在一定的量效、时效关系。2—8μmol／L浓度的As2O3可以诱导Raji细胞周期阻滞及凋亡，而1μmol／LAs2O3不诱导细胞凋亡，只是通过细胞周期阻滞作用抑制细胞增殖。结论一定浓度三氧化二砷可以抑制Raji细胞的增殖，阻滞细胞周期，诱导细胞凋亡。细胞周期阻滞伴随细胞凋亡而发生。     

硼替佐米与吡喃阿霉素联合对T细胞淋巴瘤细胞系Hut-78细胞增殖和凋亡的影响

目的 探讨硼替佐米(BTZ)与吡喃阿霉素(THP)联合对人皮肤T细胞淋巴瘤细胞系Hut-78细胞的增殖抑制作用及其机制.方法 不同浓度的BTZ、THP单药或两药联合作用于Hut-78细胞48 h后,采用MTT法检测细胞增殖抑制率,应用联合指数分析两药是否存在协同效应,在此基础上应用流式细胞术分析细胞周期分布和细胞凋亡情况,利用Western blot法检测细胞周期抑制蛋白P21的变化.结果 BTZ与THP单药对Hut-78细胞增殖具有显著的抑制作用,呈时间和剂量依赖性,两药联合时,BTZ序贯THP组的细胞增殖抑制率显著高于BTZ与THP同时组及THP序贯BTZ组(P值均＜0.01),当细胞增殖抑制率＞50%时,THP序贯BTZ呈拮抗效应,而BTZ与THP同时及BTZ序贯THP则表现为协同效应,随着增殖抑制作用的增加,仅BTZ序贯THP的协同作用更为显著.BTZ与THP单药能有效诱导Hut-78细胞凋亡,呈剂量依赖性,且BTZ序贯THP的细胞凋亡诱导作用较单药显著增强.BTZ单药可使细胞明显阻滞于G2/M期,上调P21蛋白的表达水平,序贯应用THP可显著增强BTZ对细胞周期的阻滞作用.结论 BTZ序贯THP可协同抑制Hut-78细胞增殖并诱导其凋亡,协同机制可能与序贯应用THP增强BTZ介导的P21表达上调及G2/M期细胞阻滞有关,提示BTZ与THP联合可能成为治疗T细胞非霍奇金淋巴瘤的新选择.

Abstract：

Objective To investigate the in vitro effect of bortezomib (BTZ) alone and in combination with pirarubicin (THP) on the growth inhibition of human cutaneous T-cell lymphoma cell line Hut-78.Methods Hut-78 cells were cultured with different concentrations of BTZ or THP alone and the two drugs combination for 48 h. Cell proliferation, cell cycle and apoptosis were evaluated. The cell cycle inhibitor P21 was determined by Western blot. Results BTZ or THP alone significantly inhibited the growth of Hut-78 cells in a time- and dose-dependent manner. In the combination groups, the inhibitory effect of BTZ followed by THP was the highest ( P ＜ 0. 01 ). When the inhibition rate was more than 50%, the combination index analysis showed significant synergistic if treated with BTZ followed by THP or the two at the same time, but antagonistic if treated with THP followed by BTZ. With the inhibition rate increasing, only the synergistic effect of BTZ followed by THP was further increased. The apoptosis rate of BTZ followed by THP was higher than that of single agent each(P ＜0. 01 ). BTZ alone significantly increased the proportion of cells in G2/M phase ( P ＜0. 01 ) in a dose-dependent manner and up-regulated the expression level of P21. Sequential THP notably enhanced BTZ-induced cell cycle arrest and apoptosis. Conclusions BTZ alone effectively induces growth inhibition and apoptosis of Hut-78 cells in vitro. BTZ followed by THP can synergistically enhance this cytotoxic effect. The mechanism may be that THP enhances BTZ-induced G2/M arrest and P21 up-regulation.     

抑制Skp2基因表达对NK/T细胞淋巴瘤细胞增殖和凋亡的影响

目的:研究抑制Skp2基因表达对NK/T细胞淋巴瘤细胞系HANK1细胞增殖和凋亡的影响,探讨抑制Skp2基因表达治疗NK/T细胞淋巴瘤的潜在应用价值。方法:用RNA干扰(RNA interference,RNAi)方法抑制HANK1细胞Skp2基因表达,实时定量RT-PCR和Western blot检测Skp2基因表达,MTT法检测细胞增殖,流式细胞仪检测细胞周期分布和细胞凋亡。结果:Skp2-shRNA表达载体明显抑制HANK1细胞Skp2基因表达,细胞增殖受抑,细胞周期G1期停滞,凋亡细胞增多。结论:抑制Skp2蛋白功能可能是治疗NK/T细胞淋巴瘤的好方法,值得深入研究。     

甘草次酸诱导U266细胞凋亡及对Survivin表达的影响

本研究旨在观察甘草次酸(glycyrrhetinic acid,GA)对人骨髓瘤细胞系U266增殖、凋亡及survivin表达的影响。用MTT法检测GA对U266细胞增殖活性的影响;流式细胞仪检测不同浓度GA作用于U266细胞后细胞凋亡,细胞周期的变化;扫描电子显微镜观察细胞凋亡的形态学变化;实时定量PCR检测GA对U266细胞survivin基因表达的影响。结果表明:GA对U266细胞有增殖抑制作用,其抑制作用呈现为时间和浓度依赖性;GA可以诱导U266细胞凋亡;扫描电子显微镜下可见核仁染色质边集,凝聚成块;GA可以诱导U266细胞周期阻滞于G0/G1期,表现为G0/G1期细胞比例升高,而G2/M及S期细胞比例下降。GA可以下调U266细胞survivin基因的表达,下调作用与浓度呈正相关。结论 :CA能抑制U266细胞增殖,呈浓度和时间依赖性,诱导其凋亡,其作用机理可能与细胞周期阻滞于G0/G1期和下调survivin基因表达有关。     

Human mononuclear cell modulation of endothelial cell proliferation

Endothelial cell proliferation is a histologic characteristic of several forms of nephritis characterized by infiltration of the glomerulus with mononuclear cells. To investigate the mechanism mediating this event, human endothelial cells isolated from umbilical veins and cultured in vitro were incubated with supernatants of cultured human mononuclear cells. Supernatants from mononuclear cells exerted a dose-dependent stimulatory effect on endothelial cell proliferation. The stimulatory effect of supernatant was almost entirely removed by prior depletion of mononuclear cells of monocytes by adherence, suggesting that a monocyte product was responsible for the activity. To investigate the nature of the ligand responsible, partially purified human interleukin I added to endothelial cell cultures was found to stimulate cellular proliferation.     

胰岛细胞移植过程中细胞受损的提示

背景:胰岛移植后可能发生有害的组织不相容性反应,影响细胞的存活及功能.目的:探讨胰岛细胞移植中早期胰岛细胞的损害程度及原因.方法:采用脑死亡自愿捐赠器官供者的胰腺,采用胶原酶P进行消化分离胰岛细胞,测定不同冷缺血时间下胰岛细胞损害程度.将胰岛细胞与血液进行分组培养,HLA匹配组:受者全血+胰岛细胞,受者全血+胰岛细胞+肝素:错配组:受者全血+胰岛细胞,受者全血+胰岛细胞+肝素;对照组:受者伞血+RPMI1640.观察移植早期可能出现的胰岛细胞损害.结果与结论:胰腺切取顺利,在冷缺血5 h以内胰岛细胞活性率都在80%以上,超过8 h活性胰岛细胞数量只有19%甚至更低.人胰岛暴露于未经抗凝的人血液中,胰岛将诱发一个迅速血细胞消耗.血小板、中性粒细胞和单核细胞计数显示,无论HLA错配还是匹配与对照组相比较血细胞都发牛明显的消耗:加入肝素后HLA错配组及HLA匹配组血细胞消耗反应明显减轻;HLA匹配组胰岛细胞体外培养24 h活性胰岛细胞数量高丁HLA错配组(P<0.05),说明良好组织相容性有利于胰岛细胞存活.结果提示冷缺血时间对胰岛细胞活性的影响很大,在冷缺血时间小于5 h的情况下获取的胰腺可以用于临床胰岛细胞移植的胰腺获取;移植到血液的胰岛细胞会有普遍性的炎症性损害及HLA相关性损害.     

Clear cell renal cell carcinoma

Clear cell RCC is the most common type of RCC that occurs in adults. It has the worst prognosis among the common epithelial tumors of the kidney. Histologically, a wide range of morphologic patterns can be encountered. Those cases with a multi-locular cystic architecture are considered to be a distinct subtype because of the clinicopathologic features.     

Tumor cell growth and cell kinetics

Cancer cells differ from normal cells in many ways, but most importantly by not responding to normal growth-control mechanisms. Whereas the growth and division of normal cells is carefully regulated to meet the needs of the body, tumor cells proliferate autonomously and continually, eventually interfering with and destroying the functions of normal tissue. Knowledge of molecular cell biology has grown exponentially over the last decade. Yet much remains to be understood before there can be a significant impact on our ability to design more effective therapeutic strategies for cancer patients, thereby decreasing mortality.     

急性白血病细胞粘附分子的表达

目的：探讨细胞粘附分子基因在急性白血病的发生和髓外浸润中的作用。方法：运用基因芯片和逆转录聚合酶链反应（RT－PCR）技术检测了25例急性白血病神经细胞粘附分子（NCAM）基因、细胞间粘附分子－1（ICAM－1）基因、血管细胞粘附分子－1（VCAM－1）基因的表达。结果：NCAM、ICAM－1、VCAM－1基因在急性白血病表达显著增高，并且有白血病细胞浸润患者粘附分子的基因表达高于无浸润患者，基因芯片检测结果与RT－PCR结果相同。结论：基因芯片能够为急性白血病的相关基因分析提供特异和可靠的数据。急性白血病的发病和浸润机制可能与骨髓细胞中粘附分子表达升高有关。     

嗅鞘细胞诱导树突状细胞成熟的作用研究

目的体外研究嗅鞘细胞(OEC)调节树突状细胞(DC)成熟的作用。方法从流产人胚胎和健康志愿者外周血浓缩白细胞中分别培养OEC和DC。使用DMEM/F-12(5%FBS)培养基,在24孔板中进行10~3OEC、10~3OEC+10~5DC共培养、10~3OEC/10~5DC分室培养、10~5DC或者10~5DC+1μg/mL LPS。在37℃,5%CO_2条件下孵育,48 h后,分别收集培养上清液和DC,用ELISA试剂检测上清中的TNF-α和IL-12p40的含量,流式细胞仪检测DC成熟的表面标志物CD40、CD80、CD86和HLA-DR的表达水平。结果经过10~(-5)mol/L的阿糖胞苷筛选培养12 d,NGFRp75免疫荧光检测表明OEC纯度达到90%以上,RT-PCR检测OEC表达NGF、BDNF、GDNF。OEC作用于DC能够增强DC分泌TNF-α和IL-12p40的表达,DC成熟表面标志物CD40、CD80、CD86和HLA-DR表达增加;同时,细胞因子和成熟表面标志物在分室培养中表达量增加更为显著。结论 OEC体外可诱导DC的成熟,且这种作用为非细胞接触的作用而是OEC通过分泌的可溶性分子作用结果。     

Basal cell and squamous cell carcinoma

OBJECTIVES: To describe the clinical and histologic subtypes, pathophysiology, recognition, and treatment options for basal cell and squamous cell carcinoma, and the molecular biology of sunlight-induced carcinogenesis. DATA SOURCES: Journal and review articles, research studies, textbooks, and clinical practice. CONCLUSIONS: Basal cell and squamous cell carcinoma will occur in more than one million cases annually in the United States, and are highly curable when detected and treated early. During the last decade, significant progress has been made in elucidating the molecular basis of skin carcinogenesis and in identifying newer approaches for the management and treatment of these keratinocyte cancers. IMPLICATIONS FOR NURSING PRACTICE: Nurses can play crucial roles in decreasing the morbidity and mortality from the skin cancer epidemic by identifying and referring patients with lesions suspicious for basal cell and squamous cell carcinomas.     

White cell related red cell antibodies

Occasional sera react weakly with a few red cells in the antiglobulin phase but without a recognizable pattern. We sought to identify the nature of such antibodies in 27 samples referred to our HLA laboratory for lymphocytotoxin testing. All samples were tested against a panel of 15 red cells by a capillary tube antiglobulin technique developed to conserve sera. This technique correlates well with tube antiglobulin tests, and can be performed with either fresh or thawed red cells. Of 27 sera, 14 contained anti-HLA B7, B17, or A28, since they reacted only with red cells from donors whose lymphocytes were B7, B17, or A28. Eight further sera probably contained anti-B7, -B17, or -A28, but reacted with one or two additional red cells. Two samples agglutinated all panel red cells so the presence of anti-B7, B17, or A28 could not be determined. In three additional sera, lymphocytotoxin testing suggested that specificity other than anti-B7, B17, or A28 was present. Of 27 sera containing weak unidentified red cell antibodies, 22 (81%) contain definite or probable anti-B7, -B17, or -A28. The identity of these troublesome antibodies can be determined by maintaining red cell panels of donors whose HLA phenotypes are known.