一种新的细胞周期分析方法

目的：采用DNA含量流式细胞术细胞周期分析方法,可分辨出G0/G1期、S期及G2/M期3个细胞周期群体,但无法区分G0/G1期、G2/M期细胞。方法：本研究建立的cyclin E A/DNA多参数流式细胞术。结果：可将细胞周期进行详细的分期,由传统的三分法（C0/G1期、S期以及G2/M期）为六分法（G0期、G1早期、G1晚期、S期、G2期以及M期）, 为细胞生物学研究,尤其是细胞增殖动力学提供新的,更为合理的分析方法。     

膀胱移行细胞癌组织中cyclinD1和CDK4的表达及其与临床病理的关系

目的　研究膀胱移行细胞癌组织中cyclinD1和CDK 4的表达及其与临床病理变化的关系。方法　采用免疫组化方法对 8例正常膀胱和 69例膀胱移行细胞癌组织中cyclinD1和CDK 4的表达进行观察。 结果　膀胱移行细胞癌组织中cyclinD1的表达高于正常膀胱组织 (P <0 .0 5 )。cyclinD1表达阳性者的肿瘤细胞分化较差 ,术后易复发 ,生存时间较短 ;而CDK 4的阳性表达仅与患者术后复发有关 (P <0 .0 5 ) ,与病理分级和生存时间无关 (P >0 .0 5 )。结论　cyclinD1的表达可作为判断膀胱细胞癌病理分级 ,术后复发和临床预后的指标 ,而CDK 4只能作为术后复发的参考指标。     

喉鳞状细胞癌中CyclinA、B1表达的临床意义

目的　探讨喉癌癌变过程中CyclinA和CyclinB1 表达的临床病理学意义。方法　用免疫组化检测 2 0例正常粘膜、40例不典型增生病变和 60例喉癌组织中CyclinA和CyclinB1 的表达。结果　①CyclinA和CyclinB1 阳性表达定位于细胞核。②在喉癌癌变过程中 ,喉正常粘膜、不典型增生病变和喉癌中CyclinA阳性表达率分别为 :1 0 .0 % (2 /2 0 ) ,30 .0 % (1 2 /40 )和 56 .7% (34/60 ) (P <0 .0 0 1 ) ;CyclinB1 阳性表达率分别为 :1 5 .0 % (3/2 0 ) ,35 .0 % (1 4 /40 )和 63 .3 % (38/60 ) (P <0 .0 0 1 )。③淋巴结转移组CyclinA和CyclinB1 阳性表达率明显高于非淋巴结组 (P <0 .0 5)。结论　CyclinA和CyclinB1 异常表达是喉癌发生中早期分子事件 ,并对喉癌发展起重要作用     

非小细胞肺癌中细胞周期蛋白D1与E的作用及相互关系

目的 :研究细胞周期蛋白D1与E在非小细胞肺癌 (NSCLC)发生、发展中的作用及相互关系。方法 :免疫组化方法检测 87例NSCLC肿瘤组织中细胞周期蛋白D1、E的表达及PCNA估计增殖指数 (PI) ,并将上述结果与临床病理及预后资料进行对比分析。结果 :87例NSCLC中细胞周期蛋白D1、E阳性率分别为 4 4 .8% (39/87)、4 8.3% (42 /87) ,细胞周期蛋白D1阳性组的PI值显著高于阴性组 (P〈0 .0 5 ) ,细胞周期蛋白E阳性组PI值与阴性组无显著差异 (P〉0 .0 5 ) ;细胞周期蛋白D1阳性组肿瘤直径、淋巴结转移率和生存率均与阴性组有显著差异(P〈0 .0 1、〈0 .0 5、〈0 .0 1) ,细胞周期蛋白E阳性组淋巴结转移率、临床分期和生存率均与阴性组有显著差异 (P〈0 .0 5、〈0 .0 5、〈0 .0 1) ;细胞周期蛋白D1阳性组中细胞周期蛋白E的阳性率显著高于其阴性组 (P〈0 .0 5 ) ;细胞周期蛋白D1与细胞周期蛋白E双阳性组的PI值、肿瘤直径、淋巴结转移率显著高于非双阳性组 (P值均〈0 .0 5 ) ,生存率显著低于非双阳性组 (P〈0 .0 1)。结论 :细胞周期蛋白D1、细胞周期蛋白E均参与NSCLC的发生、发展 ,并影响其预后 ,但两者在其中所起作用不同 :细胞周期蛋白D1是调节NSCLC增殖的主要因素 ,细胞周期蛋白E主要与NSCLC进展有关 ;细胞周期蛋白D1可促     

人脑胶质瘤组织中cyclin D1和cyclin E表达的研究

目的：研究人脑胶质瘤中细胞周期素(cyclin) D1和cyclin E的表达与肿瘤病理等级的关系。方法：采用免疫组织化学法检测52例人脑胶质瘤和8例正常脑组织标本中cyclin D1和cyclin E的表达。结果：人脑胶质瘤组中有29例cyclin Dl的表达。随着胶质瘤病理分级增高，高、低恶性度胶质瘤的“阳”性率和平均标记指数均正著升高，两者差异有统计学意义，P<0．05。cyclin Dl与cyclin E的平均标记指数之同呈明显正相关，Pearson相关系数r-0．644，P<0．0l。结论：cyclin Dl与cyclin E在胶质瘤中被异常表达，是肿瘤发生和恶性转化的重要促进因子。     

细胞周期分析方法

越来越多的资料显示肿瘤是一类细胞周期性疾病[1,2],从而使细胞周期分析在细胞生物学、肿瘤生物学领域显得格外重要。对于细胞增殖的细胞周期分析,就是对细胞内周期性变化的物质进行检测,以达到对细胞增殖进行检测的目的。大致可分为以下几个阶段：①显微镜下的细胞形态周期性变化分析;②细胞内DNA同位素标记并逐渐与流式细胞仪结合的分析;③通过流式细胞仪对细胞周期中合成的蛋白质进行细胞周期分析。本文着重阐述流式细胞仪对细胞进行DNA及细胞周期蛋白(cyclin)同步检测的新一代细胞周期分析方法。 1 初级的细胞周期分析方法 早些年通过显微镜对细胞的观察发现细胞是通过有丝分裂而进行增殖的,从而将细胞周期分为细胞间期和有丝分裂期,在显微镜下可以看到有丝分裂期的细胞染色体凝聚[3]。在两次有丝分裂之间的时间内除可以看到细胞体积增大外对细胞内其他变化知之很少,此为从细胞形态上对细胞周期进行分析。通过显微镜进行的细胞周期分析是极为粗略的,直到发现染色体中的DNA里携带有遗传信息才对间期有一定的了解。在细胞间期一个很短时间内通过在细胞培养液中掺入同位素标记的核苷,只在有DNA合成的细胞中才有被标记上同位素标记的核苷,从而检测到了染色体的复制,由此把间期分为3个期,即G1期(有丝分裂期与DNA复制开始的时间段),S期(即DNA合成的时间段),G2期(即S期与M期的时间段)。但是这两种方法因耗材、粗糙、效率低而无法大规模地进行应用[4]。     

1，25二羟基维生素D3对肺腺癌细胞株生长及凋亡的影响

目的：研究1，25二羟基维生素D3[1，25（OH）。VitD3]对肺腺癌细胞株A549生长及凋亡的影响。方法：应用流式细胞仪、免疫组织化学和RT-PCR等方法观察1，25二羟基维生素D3对人肺腺癌A549细胞株生长、细胞周期、调控细胞周期基因和调控凋亡基因表达水平的影响。结果：μmol／L的1，25二羟基维生素D3时A549细胞的生长有抑制作用，1μmol／L的1，25二羟基维生素D3作用后A549细胞G1期比例由59．7％增加至73．2％，P=0．001；S期比例由26．4％降至21．1％，P=0．018；细胞凋亡率由2.6％增至7．2％，P=0．015；可明显上调Fas／FasL的表达量，下调bcl-2表达量；可抑制细胞周期蛋白D1的表达量由0．762减至0．207，P=0．001；同时抑制细胞周期蛋白依赖性蛋白激酶4的表达量由0．642减至0．129，P=0．002。但较低浓度的1，25二羟基维生素队上述作用不明显。结论：1μmol／L的1，25二羟基维生素D1可通过多种途径发挥抗肺腺癌作用。     

雷帕霉素抑制人骨肉瘤MG-63细胞周期的实验研究

目的观察雷帕霉素对人骨肉瘤细胞株MG-63增殖、周期及Cyclin D1基因表达的影响,探讨雷帕霉素抑制骨肉瘤细胞周期的可能机制。方法体外培养骨肉瘤MG-63细胞,用不同浓度的雷帕霉素（1、10、25nmol／L）干预MG-63细胞。采用水溶性四唑盐WST-8比色法检测MC-63细胞增殖的变化;流式细胞仪检测MG-63细胞周期;RT—PCR及免疫细胞化学SABC法检测MG-63细胞Cyclin D1基因表达的变化。结果雷帕霉素能显著抑制MG-63细胞增殖,呈时间依赖性,差别有显著性意义（P〈0．05）;流式细胞分析显示雷帕霉素能显著抑制细胞周期,使GO／G1期细胞增多（P〈0．05）;RT—PCR检测显示雷帕霉素对MG-63细胞Cyclin D1 mRNA表达无明显影响（P〉0．05）;免疫细胞化学分析雷帕霉素能显著抑制Cyclin D1蛋白的表达,差别有显著性意义（P〈0．05）。结论雷帕霉素能下调MG-63细胞Cyclin D1蛋白的表达,抑制MG-63细胞增殖及细胞周期。     

cyclin E和cdk2在肺癌中表达及其临床意义

目的 :探讨细胞周期调控因子在肺癌发病中的作用。方法 :应用免疫组化方法检测细胞周期蛋白E(cyclinE)和细胞周期蛋白依赖性激酶 2 (cdk2 )在 38例肺癌和 6例正常肺组织中的表达状况。结果 :cyclinE和cdk2在肺癌中阳性率分别为 6 0 .5%、6 5.8% ,显著高于正常肺组织 (P <0 .0 5) ;cdk2表达与肺癌分期密切相关 (P <0 .0 5) ,而cyclinE表达与肺癌分期、淋巴结转移和病理分型均无相关性 (P>0 .0 5) ;肺癌中两者共表达率为 31.8%。结论 :cyclinE和cdk2参与肺癌的发生发展过程 ,但可能不是反映肺癌生物学特征有价值的标记物。     

细胞周期素D1与肺癌研究进展

细胞周期素D1（CyelinD1）在细胞周期的正常调控中起着关键作用。近期研究发现，在肺癌发病过程中CyclinD1基因表达失衡有不同的分子生物学机制，其表达增高的部位、水平、和方式均有重要特点。现综述CyelinD1与肺癌研究的最新进展。     

Telomeres: a Nobel Prize at the beginning… of the end

The 2009 Nobel Prize for Physiology and Medicine was awarded to Elizabeth H. Blackburn, Carol W. Greider and Jack K. Szostak for their work on telomeres and telomerase. This prize acknowledges their pionneering discoveries on chromosomal extremities. Telomeres are the nucleoproteic complexes that may be found at the ends of linear chromosomes. They are essential for genomic stability and are involved in aging and tumorogenesis.     

骨髓间充质干细胞支持造血细胞生成和分化机制的研究进展

 骨髓间充质干细胞（MSC）以其多向向中胚层起源细胞分化潜能及自我更新的干细胞特征,成为骨髓造血微环境中重要的构成组分及前体成分,参与胚胎血细胞的生成及维持造血,其本身及其衍生分化的基质细胞参与构建不同的造血龛结构,包括成骨细胞龛、基质细胞龛和脂肪细胞龛,经由细胞间多种黏附分子及相应信号通路或可溶性细胞因子直接或间接协同调控造血细胞的生成和分化,因此籍以MSC在支持造血细胞生成中发挥的作用,期待其潜在的治疗价值。     

Functional organization of the hematopoietic stem cell compartment: implications for cancer and its therapy

Recent discoveries indicate that hematopoietic stem cells have limits on their proliferative capacity and are unable to divide indefinitely. There is great heterogeneity within the compartment as to the extent of this proliferative limitation. At any given time it appears that hematopoiesis is maintained by the progeny of only a few stem cells. When these are exhausted the progeny from other stem cells take their place. The observations of proliferative limitation, heterogeneity, and clonal succession must be incorporated into any model of stem cell organization. These new discoveries and the models incorporating them have important clinical implications. They may explain the inability of normal tissues to develop drug resistance and they also offer a mechanism by which cell renewal systems decrease the development of malignancies. In the selection of chemotherapeutic agents not only the effectiveness of the drug upon the tumor must be considered, but also how specific agents affect the stem cell compartment. These data have important implications in the use of bone marrow transplantation for both malignant and nonmalignant disease.     

Glioma Migration: Clues from the Biology of Neural Progenitor Cells and Embryonic CNS Cell Migration

Neural stem cells have recently come to the forefront in neurobiology because of the possibilities for CNS repair by transplantation. Further understanding of the biology of these cells is critical for making their use in CNS repair possible. It is likely that these discoveries will also have spin-offs for neuro-oncology as primary brain tumors may arise from a CNS progenitor cell. An understanding of the normal migratory ability of these cells is also likely to have a very important impact on the knowledge of brain tumor invasion.     

MicroRNA in colorectal cancer: from benchtop to bedside

Colon carcinogenesis represents a stepwise progression from benign polyps to invasive adenocarcinomas and distant metastasis. It is believed that these pathologic changes are contributed by aberrant activation or inactivation of protein-coding proto-oncogenes and tumor suppressor genes. However, recent discoveries in microRNA (miRNA) research have reshaped our understanding of the role of non-protein-coding genes in carcinogenesis. In this regard, a remarkable number of miRNAs exhibit differential expression in colon cancer tissues. These miRNAs alter cell proliferation, apoptosis and metastasis through their interactions with intracellular signaling networks. From a clinical perspective, polymorphisms within miRNA-binding sites are associated with the risk for colon cancer, whereas miRNAs isolated from feces or blood may serve as biomarkers for early diagnosis. Altered expression of miRNA or polymorphisms in miRNA-related genes have also been shown to correlate with patient survival or treatment outcome. With further insights into miRNA dysregulation in colon cancer and the advancement of RNA delivery technology, it is anticipated that novel miRNA-based therapeutics will emerge.     

原子力显微镜下癌细胞的研究进展

癌细胞由于浸润破坏器官的结构和功能，并发生转移，而对机体产生严重影响。原子力显微镜（AFM）的出现为定量测定水溶液中单个活细胞的力学性能提供了强有力的工具。本文从四个方面阐述了AFM下研究癌细胞的进展:癌细胞力学性质、癌细胞外基质、癌细胞的迁移、抗肿瘤药物的探索，最后讨论了面临的挑战和未来的发展方向。在目前临床治疗中应用这些基本的特性可提高癌症的诊断效率、治疗水平和预防控制力，最终可以改变患者的生活。     

Molecular targets in melanoma: strategies and challenges for diagnosis and therapy

In coming years, we expect rapid advances in cutaneous melanoma diagnosis and therapy, because of the incorporation of new technologies into experimental and clinical research. Major discoveries in melanoma are often made by investigators outside the field, and the melanoma research community will need to develop a better means of incorporating these advances into their work, to capitalize on the promise they hold for patients. A far greater level of cooperation between labs and clinics will be to bring new technology-based discoveries from bench-to-bedside and back. Metastatic melanoma should become a treatable disease in the next few years, because specific inhibitors are expected for most major targets. However, major challenges lie ahead in securing funding, building infrastructure and gaining expertise in new technologies. To meet these challenges, multidisciplinary collaborations will be required all the more.     

New Therapeutic Approaches for Invasive Lobular Carcinoma

Invasive lobular carcinoma (ILC) is the second most common histological subtype among newly diagnosed breast cancers. The clinical features of ILC are related to the underlying biology of this disease, which is distinguished by loss of the cell adhesion protein e-cadherin. The resulting poorly cohesive growth pattern is responsible for the comparatively poor detection of ILC both on imaging and palpation. Recent genomic and proteomic discoveries have identified other molecular features of ILC which may lead to its future treatment with more targeted therapy. The presentation, treatment, and outcome of ILC is reviewed and future research directions are considered.     

Hedgehog signaling pathway: Development of antagonists for cancer therapy

The hedgehog pathway, initially discovered in Drosophila by two Nobel laureates, Dr. Eric Wieschaus and Dr. Christiane Nüsslein-Volhard, is a major regulator for cell differentiation, tissue polarity, and cell proliferation. Studies from many laboratories—cluding ours—eal activation of this pathway in most basal cell carcinomas and nearly one third of extracutaneous human cancers, including medulloblastomas and gastrointestinal and prostate cancers. Even more exciting is the discovery and synthesis of specific signaling antagonists for the hedgehog pathway, which have significant clinical implications in novel cancer therapeutics. This review discusses the current understanding of the hedgehog signaling pathway and its activation in human cancers. It also discusses putative and confirmed signaling antagonists and their perspectives in therapeutic applications.     

Meeting report: The international conference on tumor progression and therapeutic resistance

A multidisciplinary conference was held November 7 to 9, 2004 in Philadelphia, PA to focus on the problem of drug resistance in cancer. A great deal of knowledge is beginning to unravel the complex molecular and cellular changes associated with malignant tumor progression. With this comes many opportunities for therapeutic development. Featuring the latest tools, models, and research findings, this conference which included over 250 members of both academia and industry was a great opportunity to learn and develop new approaches and collaborations. The Keynote speaker was Dr. Robert Horvitz (Massachusetts Institute of Technology), who won the 2002 Nobel Prize in Medicine for his pioneering work on the cell death pathway in Caenorhabditis elegans. Speakers covered various aspects of tumor progression and therapy from simple models to clinical trials.