垂体瘤细胞增殖与凋亡关系的研究

目的 :探讨垂体瘤细胞增殖与凋亡的关系。方法 :应用免疫组化法和TUNEL法对 2 0例复发、2 6例未复发的人脑垂体瘤增殖细胞核抗原 (PCNA)与凋亡进行检测。结果 :复发组垂体瘤增殖细胞指数 (PCNAL1)及凋亡细胞数量较未复发组明显增高 ,两组有显著性差异 (P <0 0 5)。结论 :肿瘤细胞的增殖细胞核抗原表达指数和凋亡检测是判断垂体瘤预后的指标之一 ,有临床指导意义。     

Effects of colony stimulating factors on isolated microglia in vitro

Effects of colony stimulating factors (CSF), known regulators for cells in monocytic lineage, on isolated microglia were examined. Interleukin-3 (IL-3) induced only morphological changes in rod-shaped microglia, while granulocyte-macrophage CSF (GM-CSF) and CSF-1 induced both morphological changes and proliferation of microglia. CSF-1 also activated the enzyme activity of microglia. These observations indicated that, in terms of regulation by cytokines, microglia are similar to mature cells in monocytic lineage.Although astrocytes reportedly produce IL-3 and GM-CSF, the effects of astrocyte-conditioned medium (Ast-Sup) were different from those of either IL-3 or GM-CSF. Ast-Sup caused ameboid microglia to become ramified, and did not induce proliferation of microglia. Factors from astrocytes may have a role in the transformation of microglia which occurs in either normal developing brain or inflammation in the brain.     

人脑胶质瘤中cyclin E的表达及其对细胞增殖活性的影响

目的研究人脑胶质瘤中细胞周期素E(cyclin E)表达与胶质瘤细胞恶性程度及细胞增殖活性的关系.方法采用免疫组化方法检测52例人脑胶质瘤和8例正常脑组织标本中cyclin E和增殖细胞核抗原(PCNA)的表达水平,观察并分析各病理等级中两者表达的阳性率和标记指数.结果正常脑组织中无cyclin E或PCNA表达.随着人脑胶质瘤病理分级的增高,cyclin E的阳性率和平均标记指数均明显升高,在高、低度恶性度肿瘤之间也存在明显差异(P<0.05).双元相关性分析发现cyclin E与PCNA的平均标记指数呈明显正相关(Pearson相关系数r=0.576,P<0.01).结论人脑胶质瘤中cyclin E的表达与细胞增殖活性和病理学分级关系密切,很可能对肿瘤细胞的增殖和恶性转化产生重要的推动作用.     

原代培养多形性胶质母细胞瘤的增殖与放射敏感性

目的探讨多形性胶质母细胞瘤的增殖活性与放射敏感性,为临床治疗提供参考.方法观察了5例原代培养的多形性胶质母细胞瘤的形态学变化并绘制生长曲线.采用免疫组化链酶亲合素-生物素-过氧化物酶复合物法(SABC)检测增殖细胞核抗原(PCNA)的表达,并用四唑盐比色法(MTT)检测原代培养多形性胶质母细胞瘤以60Co 2Gy放射剂量照射后的存活分数.结果 5例原代培养的多形性胶质母细胞瘤细胞形态多样,生长曲线和增殖活性各不相同,放射敏感性也存在明显差异.结论多形性胶质母细胞瘤存在不同放射敏感性的细胞群,放射治疗应体现个体化原则.     

人脑胶质瘤中血管内皮生长因子表达与胶质瘤微血管数及肿瘤增殖活性关系的研究

目的　研究血管内皮生长因子 (VEGF)表达与胶质瘤微血管数、肿瘤增殖活性及其与胶质瘤恶性程度的相关性。方法　应用免疫组化方法检测 5 0例胶质瘤、8例正常脑组织中VEGF、血小板内皮细胞粘附分子 (CD3 1)和增殖细胞核抗原 (PCNA)的表达 ,测定其阳性细胞数和阳性血管数。结果　胶质瘤中均有VEGF、CD3 1及PCNA表达。其表达水平与肿瘤恶性程度呈显著正相关 ,其r分别为 0 .745 ,0 .765及 0 .685。结论　VEGF、CD3 1和PCNA在胶质瘤中的高表达是胶质瘤恶性表型之一 ,可作为病理诊断的补充。     

复发性脑膜瘤的细胞增殖和凋亡

目的探讨细胞增殖和凋亡在预测脑膜瘤复发方面的价值。方法应用免疫组化方法检测60例脑膜瘤的增殖细胞核抗原标记指数(PCNALI),并应用TUNEL技术检测肿瘤细胞的凋亡指数(AI),比较复发组与非复发组PCNALI、AI之间的差异。结果复发组的PCNALI(63.1%±15.3%)、AI(0.25%±0.09%)皆显著高于非复发组的PCNALI(47.7%±22.2%)、AI(0.10%±0.09%)。结论脑膜瘤的细胞增殖和凋亡与术后复发关系密切,通过检测PCNALI和AI有助于判断肿瘤的复发倾向。     

侵袭性与非侵袭性泌乳素腺瘤细胞增殖活性的差异

目的　研究侵袭性与非侵袭性泌乳素 (PRL)腺瘤细胞增殖活性的差异。方法　检测 2 6例泌乳素腺瘤中细胞核增殖抗原 (PCNA)表达 ;用流式细胞仪 (FCM )检测相应腺瘤中S期细胞比例 ,比较侵袭性泌乳素腺瘤与非侵袭性泌乳素腺瘤PCNA表达与S期细胞比例的差异。结果　侵袭性PRL腺瘤中PCNA表达的阳性指数为 76 7±1 1 1 8;非侵袭性腺瘤中PCNA表达的阳性指数为 37 1± 1 0 33;两组比较 ,差别具有显著性 (P <0 0 5) ;侵袭性PRL腺瘤S期细胞比例为 1 3 57%± 2 1 6 % ;非侵袭性PRL腺瘤为 8 97%± 2 0 7% ,两组比较差别具有显著性 (P <0 0 5)。结论　PCNA表达与S期细胞比例的检测结果是一致的 ,侵袭性与非侵袭性PRL腺瘤相比 ,细胞增殖能力较强 ,复发率高     

Differential response to acrylonitrile toxicity in rat primary astrocytes and microglia

Acrylonitrile (ACN) is a widely used chemical in the production of plastics, resins, nitriles, acrylic fibers, synthetic rubber and acrylamide. While acute high level exposures to ACN are known to be lethal, chronic low dose exposures causes glial cell tumors in rats. Recently, these glial tumors have been characterized as microglial in origin. While effects of ACN on astrocytes, the more numerous glial cell, have been investigated, the effects on microglia are unknown. This study was conducted to compare the responses of astrocytes and microglia to ACN treatment in vitro to address differential sensitivities and adaptive responses to this toxic chemical. Cell viability, ACN uptake, lipid peroxidation byproducts (F₂-isoprostanes), glutathione (GSH) levels and expression of NF-E2-related factor 2 (Nrf2) were evaluated in primary rat microglia and astrocytes following ACN treatment. Results indicate that microglia are more sensitive to ACN than astrocytes, accumulating less ACN while demonstrating higher F₂-isoprostane levels. GSH levels were up-regulated in both cell types, as a protective mechanism against ACN-induced oxidative stress, while Nrf2 levels were only induced in microglia. Our data suggest that microglia and astrocytes exhibit different sensitivities and responses to ACN, which are linked to the intracellular thiol status inherent to each of these cell types.     

IGF-1表达与人脑膜瘤细胞增殖活性关系

目的探讨IGF-1在人脑膜瘤中的表达与脑膜瘤细胞增殖活性之间的关系。方法应用免疫组化SP法检测42例脑膜瘤组织、7例正常硬脑膜中IGF-1和增殖细胞核抗原(PCNA)的表达情况,结果进行统计学分析。结果 IGF-1在正常硬脑膜组、WHO I、II和III级脑膜瘤组的表达强度不等,分别为0%、(15.7±12.8)%、(47.2±11.7)%和(62.9±12.9)%,其差异均具有统计学意义(P<0.05)。PCNA在同样四个组别中的PCNA-LI值分别为0%、9.6±5.4%、30.6±5.6%和48.7±15.1%,其差异亦均具有统计学意义(P<0.05)。相关分析结果显示:IGF-1与PCNA-LI两者呈正相关(r=0.8594)。结论 IGF-1表达强度与脑膜瘤细胞增殖活性有关,二者在脑膜瘤的发生、增殖和恶化过程中可能起协同作用,其详细机制有待进一步研究。     

Tumor cell proliferation and apoptosis in medulloblastoma

The distribution of proliferating cell nuclear antigen (PCNA)-(clone PC 10)- and Ki-67- (clone MIB-1)-positive nuclei was investigated in 60 medulloblastomas of childhood. Although the labeling index of the two markers did not coincide, both showed a wide range of parallel variations. The percentage of positive nuclei was similar in both classic and desmoplastic tumors. A variable proliferation capacity was found in the different tumor structures. Areas with neuronal and glial differentiation showed very few positive nuclei; these were very abundant in the infiltration areas, and along penetrating vessels from subarachnoidal growths. Pale islands were negative or positive only in their peripheral part. Large-cell areas were richer in positive nuclei than classic ones, accounting for their more malignant character. Hyperchromatic round nuclei, not belonging to necrotic foci and called lymphocyte-like nuclei, differently interpreted in the past, were vairiably found in every case. They are known, from previous experience, to stain orange with Acridine Orange fluorochroming, like single-stranded DNA. They were not easily distinguishable from mitoses and were stained by in situ end-labeling of DNA strand breaks, as demonstrated by incorporation of labeled nucleotides. They were regarded as possible apoptotic nuclei, representing either a peculiar type of cell death or the preservation of the cell deletion capacity, typical of the embryonal tissue of origin.Supported by AIRC, Milan and CNR, A. C. R. O.-Grant no. 9202191     

Stimulation of human fetal astrocyte proliferation by bacterial lipopolysaccharides and lipid A

Summary This report concerns the effect of bacterial endotoxin [lipopolysaccharide(LPS) and lipid A] on cultured human fetal astrocytes. Exposure to 1 g/ml LPS or lipd A caused a striking stimulation of the rate of proliferation of the cells. The effect was most pronounced with exponentially growing cultures. Stimulation was associated with enhance DNA synthesis as ascertained by [³H]thymidine incorporation. These findings at the cellular level may be of relevance in the elucidation of the effects of bacterial endotoxins on the developing human brain.     

实验性自身免疫性脑脊髓炎的小胶质细胞和星形胶质细胞反应

目的确定小胶质细胞和星形胶质细胞在实验性自身免疫性脑脊髓炎（ＥＡＥ）发病及病变发展中的作用。方法用牛脊髓髓鞘碱性蛋白（ＭＢＰ）免疫豚鼠发生ＥＡＥ,用免疫组化法观察ＥＡＥ不同病期小胶质细胞和星形胶质细胞对炎性脱髓鞘病灶的反应。结果发生ＥＡＥ的前３天,小胶质细胞即开始激活,在临床症状出现时其数量及激活程度达高峰,并持续至高峰期。恢复期数量逐渐减少,激活程度逐渐减弱。星形胶质细胞在症状高峰期开始激活并围绕在浸润细胞和病变血管周围,似有隔离小胶质细胞与病灶接触的作用,至恢复期激活明显。结论小胶质细胞激活在ＥＡＥ的发病及进展中起重要作用,而星形胶质细胞主要与疾病的恢复有关。     

白细胞介素8表达与星形细胞肿瘤增殖与凋亡的相关性研究

目的 探讨白细胞介素8(IL-8)mRNA、增殖细胞核抗原(PCNA)在星形细胞肿瘤组织中的表达及其与细胞凋亡之间的关系. 方法 应用RT-PCR方法 检测64例星形细胞肿瘤组织及10例正常大脑组织中IL-8 mRNA的表达情况.用免疫组织化学方法 检测PCNA在肿瘤组织石蜡标本中的表达.并用TUNEL法检测肿瘤细胞的凋亡. 结果 星形细胞肿瘤组织中IL-8mRNA表达与星形细胞肿瘤的恶性程度有关.随着肿瘤恶性程度的增高,PCNA表达增高,细胞凋亡指数增高.IL-8 mRNA表达与PCNA表达呈正相关关系(r=0.938,P<0.01),与细胞增殖亦呈正相关关系(r=0.907,P<0.01). 结论 IL-8mRNA表达与星形细胞肿瘤的恶性增殖呈正相关,星形细胞肿瘤通过抑制凋亡并且上调IL-8分泌促进肿瘤细胞增殖,从而导致肿瘤恶性转化.     

Immunostaining for proliferating cell nuclear antigen: its role in determination of proliferation in routinely processed human brain tumor specimens

Alcohol- and formalin-fixed, paraffin-embedded samples of 71 brain tumors (35 gliomas, 22 metastatic carcinomas, 8 meningiomas and 6 other tumors) were investigated by immunocytochemistry with three different monoclonal antibodies against proliferating cell nuclear antigen (PCNA)/cyclin (19A2; 19F4; PC10). PC10 was found to work best; it is applicable to both alcohol- and formalin-fixed tumor samples. PCNA labeling indices (LIs) were compared in the same tumors with LIs obtained by Ki-67 immunostaining of frozen sections and by in vitro incubation with bromodeoxyuridine (BrdUrd); in the latter preparations, BrdUrd LIs could be compared with PCNA LIs in the very same areas of serial sections. In gliomas, PCNA LIs were 0.7–80.2% (mean 31.7%), in metastases 0–76.0% (mean 47.8%), and in meningiomas 0–53.0% (mean 19.3%). In general, PCNA LIs were highly significantly correlated with Ki-67 LIs (P=0.0002) and BrdUrd LIs (P=0.0001). However, when tumor subgroups are considered, only gliomas show a significant correlation with Ki-67 and BrdUrd LIs. Despite this statistical correlation, PCNA expression was out of proportion to proliferation indices as determined by both other methods in almost one third of all brain tumors. Immunocytochemistry for PCNA produces a broad spectrum of staining intensity of labeled nuclei, whose number is dependent upon the sensitivity of the immunocytochemical technique used. Thus, inter-oberserver and inter-laboratory variabilities in PCNA LI determination may occur. Overlapping of PCNA LIs between tumor subgroups of varying malignancy further limits the informational value for the individual case. In some classic meningiomas, high PCNA scores do not reflect the proliferative activity of the tumor, as Ki-67 and BrdUrd LIs are very low in these cases. We conclude that PCNA immunolabeling is of limited value in the individual tumor, mainly due to overexpression in many tumors, and at present cannot be recommended to replace Ki-67 and/or BrdUrd labeling methods for routine determination of proliferative activity in human tumor specimens.Supported in part by a grant from the Oncology Commission, Medical Faculty, University of Vienna     

Reactive proliferation of astrocytes studied by immunohistochemistry for proliferating cell nuclear antigen

Astrocyte proliferation in the stab-wounded cerebral cortex of mice was studied using double immunohistochemistry for proliferating cell nuclear antigen (PCNA) and glial fibrillary acidic protein (GFAP). The number of GFAP-positive astrocytes increased markedly from day 0.5 to day 3 after stab wounding. Some GFAP-positive astrocytes in the immediate vicinity of the wound were found to be positive for PCNA. However, the maximum number of these double positive astrocytes was only 5–6% of the number of GFAP-positive astrocytes. This maximum value was observed on days 2.5 and 3. The present study revealed that astrocytes are able to reactively express PCNA, an intrinsic marker of DNA replication. On the other hand, it is suggested that the proliferation of astrocytes in the wounded cerebral cortex is limited, in contrast with their marked reactive up-regulation of GFAP.     

Colony-stimulating factors regulate programmed cell death of rat microglia/brain macrophages in vitro

Programmed cell death of activated microglia appears to be one mechanism how steady state of microglia is achieved in vivo. Programmed cell death of microglia might result either from the downregulation of microglial mitogens/survival factors or from signals which directly induce microglial cell death. To further elucidate the mechanisms regulating programmed cell death in microglia, growth factor and cytokine dependence of microglial proliferation and cell death have been examined in vitro in microglia/brain macrophage cultures established from neonatal rat brain. Microglial proliferation was assessed by PCNA labelling and DNA fragmentation by the TUNEL technique in the presence or absence of several cytokines including IL-1, IL-6, TGFβ1, TNFα, M-CSF and GM-CSF. Results of TUNEL labellings were supplemented by gel electrophoretic analysis of DNA extracted from cultured microglia which showed laddering of DNA fragments. Of all cytokines/growth factors tested, GM-CSF and M-CSF were not only the strongest microglial mitogens but, moreover, withdrawal of M-CSF or GM-CSF significantly enhanced rates of microglial cell death by DNA fragmentation. Expression of microglial growth factors, in particular colony-stimulating factors, may thus be instrumental in controlling steady states of microglia in the injured nervous system.     

环氧化酶-2在脑膜瘤中的表达及意义

目的通过检测环氧化酶-2（COX-2）在人脑膜瘤、正常脑膜组织中的表达，探讨COX-2在人脑膜瘤发生、发展过程中的作用。方法采用免疫组织化学方法检测人脑膜瘤、正常脑膜组织中COX-2的表达及其与脑膜瘤的病理分级、侵袭性、瘤周水肿等临床病理因素的关系。结果47例脑膜瘤组织中，36例有COX-2表达，阳性表达率为76．6％，正常脑膜无COX-2表达。COX-2在患者的年龄（P〉0．05）、性别（P〉0．05）之间的表达差异无统计学意义，而在肿瘤病理分级（P〈0.01）、侵袭性（P〈0．05）、瘤周水肿（P〈0．05）之间的表达差异有统计学意义。结论COX-2在脑膜瘤中普遍表达，其与脑膜瘤的病理分级、侵袭性、瘤周水肿等临床病理因素密切相关，COX-2可能在人脑膜瘤的发生发展中起重要作用。     

增殖细胞核抗原在垂体瘤的表达及临床意义

目的 探讨增殖细胞核抗原标记率(PCNA L1)作为垂体瘤增殖状态指标的临床意义。方法 35例垂体瘤标本采用免疫组织化学方法(LSAB法)。结果 全部垂体瘤PCNA均为阳性,浸润组织平均PCAN L1明显高于局限组(55．2％、28．2％),巨腺瘤平均PCNA L1明显高于大腺瘤及微腺瘤(70．2％、37．1％、24％),复发组高于初发组(63．6％、42．3％)。结论 PCNA L1垂体瘤浸润性、大小、复发有关,对判断垂体瘤侵袭性具有一定的临床意义。     

Expression of the receptor for macrophage colony stimulating factor by brain microglia and its upregulation in brains of patients with Alzheimer's disease and amyotrophic lateral sclerosis

The receptor for macrophage colony stimulating factor (CSF-1) was localized immunohistochemically in postmortem human brain tissue. Microglia constitutively expressed the receptor for CSF-1 and its expression was upregulated in lesions of Alzheimer's disease and amyotrophic lateral sclerosis. The CSF-1 mediated pathway appears to be involved in the response and activation of microglia in the central nervous system lesions.     

VEGF及PCNA表达与脑胶质瘤分级的相关性研究

目的　研究血管内皮细胞生长因子 (VEGF)与增殖性细胞核抗原 (PCNA)蛋白在各级脑胶质瘤中的表达率及与胶质瘤病理分级的关系。方法　利用免疫组化方法检测 4 5例胶质瘤手术标本中VEGF与PCNA蛋白的表达水平 ,并比较二者的相关性。结果　随着胶质瘤病理级别的升高 ,VEGF及PCNA的表达率渐之升高 ,Ⅰ Ⅳ级病理分组中 ,VEGF阳性率分别为 2 2 .2 9± 7.6 4 ,5 3.75± 10 .74 ,70 .2 7± 14 .2 7,PCNA阳性率分别为 15 .86± 4 .5 5 ,32 .4 1± 7.4 4 ,4 7.92± 10 .31,且并各级之间有显著差异 (P <0 .0 5 )。结论　VEGF与PCNA蛋白的表达对判断胶质瘤病理分级及生物学行为有一定意义。