骨巨细胞瘤体外培养及其生物学特性的观察

１９８３～１９９２年作者通过组织培养，组化及免疫组化、电镜、细胞遗传学，动物接种等方法，对骨巨细胞瘤（ＧＣＴ）的生物学特性进行研究，认为ＧＣＴ的基质细胞有两种，即间叶性基质细胞及巨噬细胞性基质细胞，巨噬细胞性基质细胞可能为许多种不同的巨噬细胞混合体，这两种类型细胞在瘤组织内互相依赖存活。在体外培养见巨噬细胞性基质细胞逐渐减少而消亡，间叶性基质细胞则在没有巨噬细胞性基质细胞的情况下也容易老化及消亡，故目前尚未有本瘤的细胞株建立。多核巨细胞是一反应性及终末性细胞，可能由于基质细胞分泌目前尚末清楚的细胞因子，吸引血中破骨细胞前身的单核巨噬细胞到瘤组织内，并促进其分化成破骨细胞样多核巨细胞。     

小鼠胸腺基质细胞的类型和分布

本文用特殊染色方法和酸性复红法对小鼠胸腺内各种基质细胞进行了鉴别,从染色特性及细胞形态,可区分出各型上皮网状细胞、保姆细胞、巨噬细胞、肥大细胞的?光镜下观察了小鼠胸腺内各种基质细胞的分布,并证实小鼠胸腺髓质内含有胸腺小体。本文就电镜所见髓质两种上皮细胞类型并结合PAS反应,对髓质内亮细胞的特性进行了讨论。     

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。     

骨巨细胞瘤体外原代培养及特性

目的观察骨巨细胞瘤体外培养细胞的形态生长特性及DNA倍体含量。方法取手术切取的骨巨细胞瘤新鲜标本8例,以组织培养法进行原代培养,观察细胞形态、生长曲线,FCM分析其倍体含量。结果骨巨细胞瘤原代培养种可见3种细胞;多核巨细胞可以长期培养。FCM分析显示:5例复发及2例原发病例是二倍体或亚二倍体,1例原发病例是非整倍体。结论单核梭形细胞是骨巨细胞瘤的主要肿瘤性成分;FCM无助于骨巨细胞瘤的预后判断。     

卵巢勃勒纳瘤5例临床病理观察与分析

目的:观察和分析卵巢勃勒纳(Brenner)瘤的临床和病理形态学特点,并阐明其诊断和鉴别诊断要点.方法:对5例卵巢勃勒纳瘤进行临床病理学、组织化学、免疫组织化学观察分析.结果:交界性瘤中可见良性瘤形态,但细胞巢增大、大小不一、排列较紧密、无巢与巢融合、无间质浸润.恶性瘤中可见交界性瘤区域,但不一定有良性瘤区域,且巢大、融合、有"出芽"现象等,并伴坏死、出血,向周围浸润或转移.结论:上皮巢和致密的纤维间质共存是勃勒纳瘤的基本结构成份.上皮巢和巢中上皮细胞形态的差别是鉴别良恶性瘤的要点.     

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.     

骨巨细胞瘤内多核巨细胞的超微结构观察

本文观察了六例骨巨细胞瘤,探讨了多核巨细胞的超微結构改变,其组织发生有迹象表明由单核基质细胞融合形成的。根据形态特征、恒定存在和生物学行为综合分析,认为多核巨细胞也是骨巨细胞瘤的实质瘤细胞。     

人骨巨细胞瘤细胞DNA含量的研究

将体外培养的人骨巨细胞瘤第四代和第五代瘤细胞,进行~3H—TdR放射自显影术和Feulgen反应,先计算出银粒标记细胞(S,G_2)的增殖比率,再用显微分光光度法测定未被标记的细胞(G_0和G_1期)的DNA含量。结果,第四代单核瘤细胞DNA含量为亚四倍体值((?)=87.36,DI=1.83),第五代单核瘤细胞DNA含量为超四倍体值((?)=118.13,DI=2.47).两代瘤细胞DNA含量经统计学处理,有显著差异(P<0.01),证实了骨巨细胞瘤细胞长期传代培养可以恶化。     

TRANSPLANTABLE MOUSE OF GLlOBLASTOMA AND ITS MORPHOLOGIC OBSERVATION

A line of cultured mous.e glioblastoma cells has been established. It is highly malignant with low differentiation and divides vigorously. Tumor tissue hemorrhages and necroses, cere- bral tumor cell infiltration is prominent. Trans- mission electronmicroscope shows morphologic irregllIarity, expanded rough endoplasmic reti- culum and free plasma polyribosomes. i The large nuclei are irregular, have membrane pores and much euchro,matin. Ilyperfunctioning nu- cleic acid metabolism is evidenced. Scanning electronmicroscope shows cell surface microvilli, filopodia and foot processes. Intercellular ad- hesive devices are connected by filopodia. Tumor cell surface ultrastructure is destroyed in the necrotic area.s.     

关于多核巨细胞发生的研究

本丈主要研究多核巨细胞的发生,采用人肾细胞株培养24小时,观察发现其中的多核巨细胞来源于巨细胞,其过程是核仁以出芽方式形成若干个新核仁,核膜内陷,包围核仁,最后核膜连接部断裂,在细胞质内形成若干个细胞核,即形成多核巨细胞,未见细胞融合。     

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.     

实验性家兔蘑菇肺的病理学观察

实验组家兔蘑菇肺的基本病变为肺的巨噬细胞性间质肺泡炎,主要在肺间质和肺泡内有大量巨噬细胞和少数多核巨细胞浸润,巨噬细胞呈弥漫性分布或聚集形成巨噬细胞结节及假结核结节。经 PAS 染色,巨噬细胞和多核巨细胞内可见 PAS 阳性红染小体。实验恢复组家兔在脱离蘑菇房后,病变可自行吸收、消散。     

CYTOLOGICAL CHARACTERISTICS OF MAIN CELLULAR ELEMENTS IN BFNIGN AND MALIGNANT GIANT CELL TUMOR OF BONE

Cytological characterist;cs of three main cellular elements iii benign and malignant giant cell tumor of bone were investigated by cytochemistry, tissue ccilture. electron microscopy and microspectropho. toinetry. The macrophage (MO), as identified by EA rosette assay, and the multinucleated giant celf (MGC) had similar enzyme activities, which differed from those of the non rosette-forming cell (NRFC). MO and MGC of malignant giant cell tumor of bone (MCCT) did not proliferate in vitro while ;ts NRFC could maintain seri81 growtfi with concomi tant formation of some multinuclear cells which are clifferent from MCC in the light and eleccron of atypical MCCs were certain aspects. Under both microscope a small number found in MGCT. The;r nii clei varied in size and shape. The NRFC in MGCT showed more prominent ritypical ultrastructural features. Part of the NRFCs had aneuploid DNA content above 4C (-4C), which is generally consi dered as the evidence of rnalignancy. Some nuclei i" the MGCs of MGCT contained }40 DNA con tent, but not those in MGCs of benign giant cell tumor of bone. Usually, no aneupaoidy was detected in MO of all the samples with the exception of one MGCT sample. in which l O% M¢ contains :4C DNA content. Iri the in virro labelling, :II-TdR labelled NRFC were in varying percentage, but labelled MO and MGC were infrequently found. The circumstan tial evidences suggest a close relationship exists be. tween M¢ and MGC: but NRFC presents neoplastic featurea. The atypical giant cells in MGCT might be of tumor origin.     

MALIGNANT BONE FIBR.OUS HISTIOCYTOMA REPORT OF 3 CASES

Three ca.ses of malignant bone fibrous his- tiocytoma are reported. They were identified on the basis of charact.eristic histopathologic fea.- tures, i.e. neoplastic histiocytes, fibroblast-like cells and multinuclear or mononuclear tumor giant cells, generally appearing very pleomorphic and bizarre and fibroblastic cells and collagen fibers arranged in storiform or matted fashion, as well as active phagocytosis, prominent mitosis and sprinkling of lymphocytes and plasma cell infiltration. The storiform pattern is considered the most characteristic feature, which may be of great help in differential diagnosis. The clinical manifestations, roentgenologic pictures and sur- gery results in 3 cases are also described.     

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     

子宫内膜间质肿瘤临床病理研究(A1AT及A1ACT检测意义探讨)

子宫内膜间质肿瘤常见为低度恶性内膜间质异位症;与高度恶性间质肉瘤主要不同点在于前者瘤细胞异形性小,核分裂相少、生长缓慢,复发及转移发生较晚,预后较好。由于两者预后有显著差别,必须严格掌握诊断标准,注意区分其恶性程度。免疫组织化学PAP法显示内膜间质肿瘤A1AT及A1ACT多阳性,与平滑肌肉瘤鉴别,具有重要参考价值。     

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。     

骨巨细胞瘤Jaffe分级的PCNA评价

目的观察骨巨细胞瘤Ｊａｆｆｅ组织分级与瘤细胞增殖的关系。方法应用免疫组化ＡＢＣ法对２７例不同组织级别的骨巨细胞瘤进行增殖细胞核抗原（ＰＣＮＡ）检测。结果ＰＣＮＡ的阳性反应仅见于基质细胞的胞核中，多核巨细胞呈阴性反应。不同组织级别的骨巨细胞瘤之间，ＰＣＮＡ阳性检出率无显著差别（Ｐ＞０．０５）。结论骨巨细胞瘤Ｊａｆｆｅ分级系统与肿瘤细胞的增殖情况无关；多核巨细胞非肿瘤细胞。     

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

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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.