骨巨细胞瘤中p53基因的异常

应用免疫组化及分子杂交方法研究了１０例骨巨细胞瘤及４例胎儿骨标本。大部分多核巨细胞（ＭＧＣ）及约２０％～４０％的单核基质细胞（ＳＴＣ）对抗巨噬细胞单抗标记呈阳性，胎儿骨中破骨细胞对抗巨噬细胞单抗标记亦呈阳性。在抗ｐ５３蛋白单抗染色中，３０％～６０％的ＳＴＣ标记阳性。ＭＧＣ呈两种标记反应，体积大、核多的ＭＧＣ呈阴性，体积小、核少的ＭＧＣ呈阳性。ＤＮＡ分子杂交发现有１例ＧＣＴ存在部分基因片断缺失。     

骨巨细胞瘤中的多核巨细胞体外骨吸收研究

目的：比较破骨细胞与骨巨细胞瘤中多核巨细胞的特点，明确后者的性质和来源。方法：用倒置相差显微镜观察体外培养的多核巨细胞的一般形态及降钙素对它的影响；用骨片与多核巨细胞共同培养法观察多核巨细胞的体外骨吸收功能，用扫描电镜观察骨吸收陷窝，Ｇｏｍｏｒｉ染色观察多核巨细胞的酸性磷酸酶活性。结果：倒置相差显微镜下可见多核巨细胞胞核较多（２０个以上），胞浆周边不规则，有伪足样突起；胞浆内可见较多大小不等的空泡；降钙素（１００μｇ·Ｌ－１）可抑制多核巨细胞的伪足样运动；多核巨细胞与灭活的骨片共同培养时可见骨吸收陷窝形成，扫描电镜下可见吸收陷窝底面有原纤维；Ｇｏｍｏｒｉ染色时可见多核巨细胞的酸性磷酸酶呈阳性。结论：证实了多核巨细胞具有破骨细胞的形态特征与骨吸收功能，可能来源于骨髓的破骨细胞前体细胞     

骨巨细胞瘤中多核巨细胞的纯化及其性质的探讨

目的 :利用酶消化方法对骨巨细胞瘤中多核巨细胞进行纯化 ,以弥补破骨细胞获取量少的问题 ,为骨质疏松的体外研究提供丰富的细胞来源。并用免疫组化的方法对骨巨细胞瘤中多核巨细胞的性质和来源进行探讨。方法 :体外分离培养 8例骨巨细胞瘤的多核巨细胞 ,在培养 2 0h后 ,用 0 .5 g·L-1胰蛋白酶 / 0 .2 g·L-1EDTA联合消化的方法去除贴壁能力较弱的单核基质细胞。将分离培养的骨巨细胞瘤多核巨细胞与牙磨片共培养 ,观察骨巨细胞中多核巨细胞的噬骨能力。并用破骨细胞特异表达的空泡型质子泵、Ⅱ型碳酸酐酶、组织蛋白酶K、基质金属蛋白酶 9对多核巨细胞进行免疫组化染色和TRAP染色。结果 :利用酶联合消化的方法可以获得较高纯度的多核巨细胞 ,纯化率达 85 %。骨巨细胞瘤中的多核巨细胞对破骨细胞所特异表达的空泡型质子泵、Ⅱ型碳酸酐酶、组织蛋白酶K、基质金属蛋白酶 9呈强阳性表达 ,TRAP染色阳性 ,体外培养具有噬骨能力。结论 :利用 0 .5 g·L-1胰蛋白酶 / 0 .2g·L-1EDTA消化的方法可获得较高纯度的多核巨细胞 ,可为破骨细胞体外研究提供丰富的细胞来源。骨巨细胞瘤中的多核巨细胞在功能表达上与破骨细胞相似 ,它可能来源于病变中圆形单核基质细胞的融合     

肺原发性巨细胞型恶性纤维组织细胞瘤（附一例报告）

报道1例肺原发性巨细胞型恶性纤维细胞细胞瘤。肉眼见右肺下叶两个分界清楚的结节,切面呈灰黄、灰红色,有骨化、灶性出血和坏死。镜下见弥漫分布的瘤细胞呈圆形、椭圆形,部分为梭形,并形成典型的车辐状结构,瘤组织内可见大量多核瘤巨细胞或破骨细胞样多核巨细胞,肿瘤的边缘见反应性骨组织形成。瘤组织中可见零星分布的凋亡多核巨细胞。免疫组化瘤细胞对vimentin、alpha-1-antitrypsin、lysozyme和mac387呈阳性,但对cyto-keratin、actin、S－100、NSE和NF均呈阴性。肺原发性巨细胞型恶性纤维组织细胞瘤非常罕见,临床治疗主要采取根治性手术切除。     

骨巨细胞瘤中p53基因的异常

应用免疫组化及分子杂交方法研究了１０例骨巨细胞瘤及４例胎儿骨标本。大部分多核巨细胞及约２０％－４０％的单核基质细胞对抗巨噬细胞单抗标记呈阳性，胎儿骨中破骨细胞对抗巨噬细胞单抗标亦呈阳性。在抗Ｐ５３蛋白单抗染色中，３０％－６０％的ＳＴＣ标记阳性。     

破骨细胞在口腔正畸领域中的研究现状

来源于单核巨噬系统的造血干细胞,在巨噬细胞集落刺激因子、粒细胞巨噬细胞集落刺激因子以及白介素等的诱发下演变成为破骨细胞.破骨细胞是进行骨吸收的主要效应细胞,是一个高度分化的多核巨细胞,直接参与骨吸收.     

骨巨细胞瘤中PCNA、p16、Rb与bcl-2基因表达及凋亡的检测研究

目的：探讨骨巨细胞瘤（ＧＣＴ）的细胞学性质、肿瘤发生以及生物学行为。方法：用免疫组化法检测了５１例ＧＣＴ中ＰＣＮＡ、ｐ１６、Ｒｂ和ｂｃｌ－２蛋白的表达及用ＴＵＮＥＬ法检测其凋亡细胞。结果：全部标本ＰＣＮＡ阳性表达，其中典型多核巨细胞未见着色；ｐ１６、Ｒｂ和ｂｃｌ－２在ＧＣＴ中的阳性率分别为９２２％、９６１％和３３３％。Ｊａｆｅ分级间ＰＣＮＡ的表达具有显著性差异；ｐ１６和Ｒｂ在ＧＣＴ中的表达具有一定的负相关；ｂｃｌ－２在Ⅰ与Ⅲ级及Ⅰ与Ⅱ＋Ⅲ级间表达差异有显著性。ＧＣＴ的凋亡率（ＡＩ）为０８６％，多核巨细胞凋亡偶见。ＧＣＴ的ＡＩ与ｂｃｌ－２的表达呈显著负相关，ｂｃｌ－２表达与否的ＡＩ之间有显著的差异。结论：Ｊａｆｅ分级可以反映出ＧＣＴ内的增殖状态；多核巨细胞是一种静息细胞；ｂｃｌ－２在ＧＣＴ发展中可能起一定的作用；联合检测ＰＣＮＡ和ｂｃｌ－２可以作为判断其生物学行为的指标。     

骨巨细胞瘤中多核巨细胞的来源

目的:研究骨巨细胞瘤中多核巨细胞的来源,探讨骨巨细胞瘤是破骨细胞或者是肿瘤细胞。方法:本次试验标本30例为解放军222医院病理科经病例证实的手术标本。采用链菌素亲生物素一过氧化物酶连接法(sp)免疫组化技术。抗体PCNA、P53、P21、CD68、AACT以及SP染色试剂均为即用型,阳性判断以细胞浆、细胞膜或细胞核着棕色颗粒为阳性反应,以缓冲液PBS代替一抗作阴性对照,确定骨巨细胞瘤中多核巨细胞的来源。结果:30例骨骨细胞瘤中的多核巨细胞均不表达PCNA、p53及c-er Bb2、有8例表达p21,其包浆课件棕褐色的阳性颗粒,所有标本中的巨细胞均呈现CD68强阳性,棕褐色的颗粒绝大多数位于细胞浆中,少数多核巨细胞呈Vimentin阳性反应,还有一部分多核巨细胞呈Lysozyme中性或阳性反应。结论:巨细胞瘤中的多核巨细胞与破骨细胞在形态学及酶组织化学与免疫组织化学与破骨细胞相似。多核巨细胞具有一定的破骨能力,由此可见,多核巨细胞是破骨细胞,属于终末细胞,而不是肿瘤细胞。     

骨巨细胞瘤中的多核巨细胞体外骨吸收研究

目的：比较破骨细胞与骨巨细胞中多核巨细胞的特点，明确后的性质和来源。方法：用倒置相差显微镜观察休外培养的多核巨细胞的一般形态及降钙素对它的影响，用骨片与多核巨细胞共培养法观察多核巨细胞的体外骨吸收功能，用扫描电镜观察同吸收陷窝，Ｇｏｍｏｒｉ染色观察多核巨细胞的酸性酸酸酶活性，结果：倒置相差显微镜下可见多核巨细胞核较多（２０个以上）胞浆周边不规则，有伪足样突起，胞闪内可可见较多大小不等的空泡；降钙     

骨巨细胞瘤的细胞生物学

目的 探讨骨巨细胞瘤中多核巨细胞和单核基质细胞的起源及在肿瘤中的作用,方法 应用免疫组织化学方法检测50例骨巨细胞瘤手术标本的增殖细胞核抗原（PCNA）、巨噬细胞抗原（CD68）、波形蛋白、抗糜蛋白酶（AACT）、抗胰蛋白酶（ACT）以及溶菌酶的表达和分布状况。结果 多核巨细胞主要表达CD68、AACT、ACT以及溶菌酶,无一表达PCNA;单核基质细胞可分为两种类型,以纤维母细胞型占优势,主要表达波形蛋白和PCNA;组织细胞型主要表达CD68、AACT、ACT以及溶菌酶。结论 多核巨细胞并非肿瘤细胞,它可能来自单核巨噬细胞系统,单核基质细胞中的纤维母细胞型为肿瘤的主要增殖成分,可能来自间充质。     

人骨巨细胞瘤细胞系GCT-0404的建立和生物学特性

目的:研究骨巨细胞瘤细胞体外培养的生物学特性,并建立人骨巨细胞瘤细胞系. 方法: 采用原代组织块培养法培养骨巨细胞瘤手术标本,对存活细胞进行形态学观察、免疫组织化学染色、细胞周期检查、核型分析、裸鼠移植. 结果: 建立人骨巨细胞瘤细胞系GCT-0404,其形态学表现、免疫组织化学染色均符合骨巨细胞瘤纤维母细胞样基质细胞的特征. 经过近1 a的体外培养,现已传代100次,细胞倍增时间39.7 h,细胞周期测定G1期为67.5%,G2期为8.9%,S期为23.6%. 染色体具有三倍体核型. 裸鼠移植成瘤率100%,无支原体污染. 结论: 人骨巨细胞瘤细胞系GCT-0404可以用于对骨巨细胞瘤的研究.     

Effect of cytokines on in vitro bone resorption by cells isolated from giant cell tumor of bone

ＯｂｊｅｃｔｉｖｅＴｏｉｎｖｅｓｔｉｇａｔｅｔｈｅｅｆｅｃｔｏｆｃｙｔｏｋｉｎｅｓｏｎｉｎｖｉｔｒｏｂｏｎｅｒｅｓｏｒｐｔｉｏｎｂｙｃｅｌｓｉｓｏｌａｔｅｄｆｒｏｍｇｉａｎｔｃｅｌｔｕｍｏｒｏｆｂｏｎｅ．ＭｅｔｈｏｄｓＭｏｎｏｎｕｃｌｅａｒｓｔｒｏｍ...     

应用单抗GCF-5观察骨巨细胞瘤细胞成分

采用免疫细胞化学和免疫电镜方法,以我室制备的抗骨巨细胞瘤(GCT)肿瘤细胞的单克隆抗体GCF-5,对41例GCT及其它肿瘤细胞进行观察,结果表明:41例中的35例GCT标本与GCF-5结合呈阳性反应;除1例骨肉瘤细胞系(OS-732)细胞为阳性反应外,其它骨肿瘤均为阴性反应。经免疫金染色后电镜下,在阳性细胞表面可见金颗粒,证明GCF-5抗体是抗细胞表面抗原的单克隆抗体,GCF-5与GCT中部分基质细胞(STC)结合,除与一些双核细胞和核数少的多核巨细胞(MGC)结合外,与绝大多数的MGC不发生反应,但能与GCT体外培养、多次传代后的MGC发生反应。均支持本作者以前的观点,即GCT中的MGC与STC各自包含两种截然不同的细胞成分:肿瘤细胞和与肿瘤免疫有关的细胞,仅肿瘤细胞成分能在体外培养中生长、增殖。     

Apoptosis in giant cell tumors of bone

Although giant cell tumor of bone (GCT) is characterized by the extensive multinucleated giant cells among mononuclear stromal cells, proliferation of these cells and multinucleation are not without limit in certain cases. Few studies on oncogenesis of GCT have focused on the negative growth control, including growth arrest and apoptosis. The purpose of this study was to investigate the mechanism of cell death in multinucleated giant cells and stromal cells of GCT. In this study, we have demonstrated that GCT cells can undergo apoptosis. The cells in surgical specimen were positively stained in situ nick end labeling methods, and electron micrographs showed the morphological changes associated with apoptosis in some of stromal cells and multinucleated giant cells. A candidate responsible for this apoptosis was then examined using cultured GCT cells. We focused on Fas that is a major trigger of apoptosis. Cultured GCT cells expressed detectable amount of Fas on their surface. Although GCT cells did a little undergo apoptosis following treatment with anti-Fas alone, combination treatment with cyclohexamide led to an increase in apoptosis of the GCT cells. These data suggested that the sensitizing activity of cyclohexamide on anti-Fas mediated cytotoxicity could happen in vitro.     

uPA-uPAR系统对骨巨细胞瘤细胞p44(MAPK)信号转导的影响

目的研究uPA／uPAR系统对骨巨细胞瘤细胞MAPK信号转导的影响。方法分离并培养骨巨细胞瘤细胞，用免疫组化检测骨巨细胞瘤组织中uPAR的表达水平，用免疫共沉淀方法检测外源激活或阻断uPA—uPAR对骨巨细胞瘤细胞信号转导通路的p44（MAPK）蛋白磷酸化水平的影响。结果仅有单核基质细胞可以在体外培养中长期生存；在骨巨细胞瘤组织中uPAR主要表达在部分单核基质细胞和一些多核巨细胞的胞膜上；将uPA—ATF加入培养的骨巨细胞瘤细胞后，细胞信号通路上的p44蛋白磷酸化水平明显增高。用uPAR抗体处理后，细胞p44蛋白磷酸化水平明显降低。说明uPA—ATF具有细胞信号转导活性，该活性受uPAR拮抗剂的影响。结论本实验检测到uPA—uPAR系统是通过p44（MAPK）信号转导通路转导信息，从而调节骨巨细胞瘤细胞增殖、分化及其他生物学行为。     

骨巨细胞瘤的诊断与治疗

骨巨细胞瘤(GCT)是一种高侵袭性良性骨肿瘤,主要发生于年轻人中,骨端发病典型的影像学是骨端完全溶骨的膨胀性改变,多不伴有骨膜反应及软组织包块;CT、磁共振可以很好地显示肿瘤的侵及范围;X线平片最具诊断意义。圆形及卵圆形基质细胞及多核巨细胞是GCT的基本结构成分。刮除术是治疗GCT的基本外科手段,但其复发率较高。行刮除术的同时应用高速磨钻处理髓腔或用液氮冷冻、石碳酸烧灼、50%氯化锌液体浸泡以及放疗等辅助方法处理刮除后的病灶,可明显降低复发率。     

IN VITRO TISSUE CUL,TURE STUDY ON GIANT CELL TUMOR OF BONE

In vitro tissue culture of 22 giant cell tumors (GCTs) of the bone in 19 cases was undertaken. Primary growth was successful in all. 11 were Grade I tumors, 10 Grade n and l Grade III. The average survival of the 3 grades of GCTs was in order 167.3, 219 and 625 days. Of these, 6 were postoperative recurrent GCTs, their life span ranging from 198 t0 6.25 days. It is apparent that the higher the pathologic grade of GCT, the longer the survival of cultured cells. Recurring tumors also manifested more active growth po- tential. This lends support to the view that the existing pathologic grading remains valuable for clinical reference. Multinucleated giant cell usually survived 3 weeks, occasionally 3 months. Growth of the stromal cells maint.ained from 5 weeks t0 8 months. The circumstantial evidence indicates that the stromal cells, not the giant cells, should be considered as the offending neoplastic invader.     

CELLULAR ELEMENTS IN GIANT CELL TUMOR OF BONE FEULGEN-DNA QUANTITATION, 3H_TdR INCORPORATION AND ULTRASTRUCTURE OBSERVATION

Anti-erythrocyte rosette assay was used to sub divide stromal cells (StCs) of cultured giant cell tumor (GCT) of bone into two groups, the rosette forming cells (RFCs) and non-rosette forming cells (NRFCs). Characteristics of Feulgen-DNA content, 3H.TdR uptake and ultrastructure of different cellular elemcnts in GCT were then investigated by micro- spectrophotometry, autoradiography and both TEM and SEM. Two types of StCs and multinucleated giant cells (MGCs) showed 2C DNA stem line, which contrasted strikingly with aneuploid DNA in osteosarcoma cells. Autoradiography revealed that the labelling index of NRFCs increased with the time of 3H.TdR incorporation in vitro, while RFCs and MGCs were scarcely tagged. The two types of StCs were distinctly different in both surface and intra- cellular structures. The phagocytic function and sur- face appearance of RFCs resembled those of ma crophages, and no polymorphism was found in RFCs. These facts suggest that NRFCs are the neoplastic element in GCT, whereas RFCs are the end-stage cells and possibly the macrophages related to tumor immunity.     

CONTRAST STUDY ON MULTINUCLEATED GIANT C,FLLS IN GIANT CELL TUMORS OF BONE AND OTHER MULTINUCLEATED CELLS

By means of tissue culture, electron microscopy, cytochemistry, indirect immunofluorescence and immunohistochemistry, the multinucleated giant cells(MGCs) in 12 giant cell tumors of bone (GCT) were studied in contrast with osteoclasts (OCs), foreign body giant cells (FGCs) and inflammatory giant cells (IGCs). The findings in the majority of MGCs were identical with those in OCs, suggesting that they most probably derived from the same precursor. Continuous in vitro culture revealed two kinds of MGCs, which were designated preliminarily as short-lived MGCs and long-lived MGCs for their difference in morphology and in several biological features, which suggests two kinds of MGCs exist in GCT. We conclude that the short-lived conform to the typical MGCs known generally to the pathologists, while the long-lived are deemed to be closely related to the neoplastic elements of the tumor.     

ULTRASTRUCTURAL INVESTIGAT10N OF GIANT CELL TUMOR OF BONE

The surface and internal ultrastructure of multinuclear giant cells and stromal cells are studied in 3 cases of giant cell tumors of bone by means of scanning electronmicroscope and transmission electronmicroscope. The fibroblast- like type of stromal cells was the principal stro- mal cell and has the function of forming collagen. Like the fibroblastic type of stromal cells, the multinuclear giant cells can also be deemed as true tumor cells. Various surface ultrastruct- ural configurations of the multinuclear giant cells can be attributed more to heightened me- tabolism than to manifestation of malignancy.