NSE和嗜铬素A在胃肠胰神经内分泌细胞和肿瘤中…

应用免疫组化方法，观察ＮＳＥ和嗜铬素Ａ在胃肠胰神经内分泌细胞和肿瘤组织中的表达。结果发现：ＮＳＥ在肿瘤组阳性率为９０．９％，嗜铬素Ａ为６０．６％；而在正常细胞组，ＮＳＥ阳性率３２．４％，嗜铬素Ａ为１００％，我们认为ＮＳＥ和嗜铬素Ａ对胃肠胰神经内分泌肿瘤的诊断都具有特殊的意义，联合应用能提高其诊断率。     

嗜铬素A阳性细胞在结肠癌组织中的表达

应用免疫组化方法检测嗜铬素Ａ阳性细胞在结肠癌组织中的表达。结果发现、嗜铬素Ａ阳性细胞在结肠癌组织中的表达为３５．４％，在正常结肠粘膜中内分泌细胞的表达率为１００％，与ＮＳＥ免疫组化染色和常用的Ｇｒｉｍｅｌｉｕｓ嗜银染色相比，嗜铬素Ａ免疫组化染色的阳性率明显高于ＮＳＥ染色和Ｇｒｉｍｅｌｉｕｓ嗜银染色。因此认为嗜铬素Ａ免疫组化染色可作为检测结肠癌组织中伴内分泌细胞分化的一种可靠指标。     

ELISA法检测肿瘤相关抗原神经元特异性烯醇化酶的方法学评价

对肿瘤相关抗原神经元特异性烯醇化酶(NSE)ELISA检测方法进行综合评价.选择临床诊断为小细胞肺癌、非小细胞肺癌及神经母细胞瘤等肿瘤患者、其它非肺癌肿瘤患者、良性疾病患者及健康人样本共223例,进行盲法比较.本方法的灵敏度为60.2%,特异性为93.8%,阳性预测值为87.5%,阴性预测值为76.7%.NSE在各入选人群中的阳性率分别为:小细胞肺癌组84.6%,非小细胞肺癌组47.6%,肺癌组总阳性率为58.4%,神经母细胞瘤组阳性率为100%,非肺癌肿瘤组的阳性率为13.3%,良性疾患组阳性率为0%,健康人群组阳性率为0%.中国正常人群cut-off值为7.78μg/L,与试剂盒提供的cut-off值相似.NSE在小细胞肺癌及神经母细胞瘤患者中有很高的阳性率,尤其是对于神经母细胞瘤有很高的临床意义.NSE在正常健康人及良性疾病患者、非肺癌肿瘤患者中的阳性率较低,检查NSE有助于肿瘤的鉴别诊断.     

INSM1在胃肠胰神经内分泌肿瘤诊断中的价值

目的探讨胰岛素瘤相关蛋白1(insulinoma-associated protein 1,INSM1)在胃肠胰神经内分泌肿瘤(gastroenteropancreatic neuroendocrine neoplasms,GEP-NEN)中的表达及其在诊断中的应用价值。方法应用免疫组化En Vision法检测48例GEPNEN和40例胃肠胰非神经内分泌肿瘤中INSM1、Cg A、Syn和CD56的表达,并分析其与GEP-NEN临床病理特征的关系。结果48例GEP-NEN中,INSM1的阳性率为81. 3%,而Cg A、Syn和CD56的阳性率分别为50%(24/48)、89. 9%(43/48)和66. 7%(32/48),INSM1、Cg A、Syn和CD56在胃肠胰非神经内分泌瘤中的阳性率分别为0 (0/40)、10. 0%(4/40)、12. 5%(5/40)和10%(4/40),INSM1、Cg A、Syn和CD56在GEP-NEN中的表达均明显高于胃肠胰非神经内分泌瘤中的表达(P 0. 05);INSM1主要表达于食管、胰腺和结直肠神经内分泌肿瘤中,与Cg A、Syn和CD56表达不同,INSM1表达与GEP-NEN分级无关(P 0. 05)。INSM1对GEP-NEN诊断的灵敏度高于Cg A,特异度与Cg A、Syn和CD56差异无统计学意义(P 0. 05)。结论INSM1属于新的神经内分泌标志物,可与Cg A、Syn和CD56联合检测用于GEP-NEN的诊断。     

NSE,CgA,Syn在神经内分泌肿瘤诊断中的应用

目的：探讨神经内分泌肿瘤诊断中神经元特异性烯醇化（ＮＳＥ），嗜铬素Ａ（ＣｇＡ），突触素（Ｓｙｎ）的应用价值。方法：应用免疫组化ＬＳＡＢ法对９９例不同部位及激素活性的神经内分泌肿瘤进行ＮＳＥ，ＣｇＡ，Ｓｙｎ标记。结果：３种标记物在神经内分泌肿瘤中的阳性率分别为ＮＳＥ９４．９％，ＣｇＡ７０．７％，Ｓｙｎ７５．８％。３种标记物共同表达率为５８．６％，高于单项（１５．２％）和双项（２５．３％）标记物表达率     

乳腺神经内分泌型导管内癌

目的对乳腺神经内分泌型导管内癌（E-DCIS）的临床病理特点、预后和鉴别诊断进行探讨。方法用光镜、免疫组织化学EnVision法行嗜铬素A、突触素和神经元特异性烯醇化酶（NSE）染色和消化PAS、消化阿辛蓝和嗜银染色,对18例具有E-DCIS特征的乳腺癌进行观察。结果具有E-DCIS特征的乳腺癌具有以下特点：（1）好发于老年女性,平均年龄71岁。最常见的临床症状为乳腺肿块或乳头溢液。（2）E-DCIS呈导管内肿瘤细胞的膨胀性生长,在肿瘤周边常可见导管内乳头状瘤。（3）肿瘤细胞呈多边形,卵圆形或梭形,胞质丰富,嗜酸性或细颗粒状。细胞核往往只有轻一中度异型,消化PAS或AB染色显示细胞内或细胞外存在黏液,有些肿瘤细胞呈印戒细胞样。（4）〉50％的肿瘤细胞表达嗜铬素A、突触素和NSE中的至少两种,部分病例CD56和CD57染色阳性。（5）E-DCIS中常可见到肿瘤细胞向邻近导管的派杰样扩散,且在膨胀性生长的导管内不存在肌上皮成分。这两点有助于E-DCIS与导管上皮增生的鉴别。结论E-DCIS是一种低度恶性的乳腺导管内癌,有其独特的组织形态、免疫组织化学特征,应作为一种独立的导管内癌类型加以认识。     

肺癌组织中神经内分泌细胞的临床病理意义

用NSE和嗜铬颗粒素抗体对40例不同类型肺癌做了免疫组化染色,以观察其与各种肺癌的分化,间质纤维组织,淋巴细胞及核分裂象的关系。40例肺癌中鳞癌21例,腺癌10例,小细胞癌7例,类癌2例,发现鳞癌中NSE和嗜铬颗粒素阳性率均为76.2％,而且多分布于中一高分化,间质纤维组织多,淋巴细胞多和核分裂象少的区域。腺癌的NSE阳性率为60％,嗜铬颗粒素阳性率为30％,多分布于低分化区,与间质纤维组织和淋巴细胞关系不密切。小细胞癌和类癌的NSE和嗜铬颗粒素均为阳性。结果可见NSE和嗜铬颗粒素在肺鳞癌的分化分级中有其参考意义,是否对预后有意义有待进一步追随病例证实。     

中国胃肠胰神经内分泌肿瘤病理学诊断共识

神经内分泌肿瘤(neuroendocrine neoplasm)是一组起源于肽能神经元和神经内分泌细胞的异质性肿瘤,可发生于全身许多器官和组织,包括胃肠道、胰腺、胆管和肝、支气管和肺、肾上腺髓质、副神经节、甲状腺、甲状旁腺以及其他部位的神经内分泌细胞,其中胃肠胰神经内分泌肿瘤最常见,约占所有神经内分泌肿瘤的55%～70%.流行病学研究显示,肺和胃肠胰神经内分泌肿瘤的发病率为5.25/10万,比30年前增高约5倍[1].发病率的增高固然与临床诊断手段的进步有关,但实际发病率也的确在增加.     

肾上腺肿瘤中A103、Inhibin α和c-erbB-2的表达

目的 研究A103、Inhibin α和c-erbB-2在肾上腺肿瘤中的鉴别诊断价值。方法 应用免疫组织化学Power Vision System法检测48例肾上腺肿瘤A103、Inhibin α和c-erbB-2的表达。结果 A103免疫组化染色阳性率分别为肾上腺皮质腺瘤76．5％(13／17)，肾上腺皮质腺癌50．0％(4／8)。Inhibin α免疫组化染色阳性率分别为肾上腺皮质腺瘤82．4％(14／17)，肾上腺皮质腺癌75．0％(6／8)。A103在肾上腺皮质肿瘤中呈胞质颗粒状阳性，主要见于肿瘤的透明细胞区域。Inhibin α呈胞质或胞膜阳性，可见于肿瘤的透明细胞和颗粒细胞区域。A103和Inhibin α两种抗体在肾上腺皮质腺瘤表达率94．1％(16／17)，在肾上腺皮质腺癌表达率为87．5％(7／8)。A103和Inhibin α在肾上腺嗜铬细胞瘤均呈阴性表达。c-erbB-2表达率：肾上腺皮质肿瘤为64．0％(16／25)，肾上腺嗜铬细胞瘤为17．4％(4／23)。c-erbB-2呈胞质颗粒状或弥漫阳性。在所有肿瘤中未观察到特异性的胞膜阳性反应。结论 A103和Inhibin α联合应用在肾上腺皮质肿瘤和嗜铬细胞瘤的鉴别诊断中具有较高价值。     

胃粘液癌和普通型胃部中P53蛋白,CgA和Syn的表达

目的：研究胃癌伴神经内分泌（ＮＥ）分化时的病理特征。方法：采用免疫组化Ｓ－Ｐ法及ＡＢ－ＰＡＳ染色技术在１００例胃癌中对比观察普通型与粘液型癌Ｐ５３表达与伴ＮＥ分化的关系。结果：１００例胃癌中ＮＥ分化标记嗜铬素Ａ（ＣｇＡ）和（或）突触素（Ｓｙｎ）阳性率为４２．１００％，其中ＣｇＡ阳性率３６．００％，ＣｇＡ与Ｓｙｎ同时阳性率１０．００％。Ｐ５３阳性率为５０．００％，普通型与粘液型癌中Ｐ５３表达差异无显     

Gastroenteropancreatic neuroendocrine tumors. A histochemical and immunohistochemical study of epithelial (keratin proteins, carcinoembryonic antigen) and neuroendocrine (neuron-specific enolase, bombesin and chromogranin) markers in foregut, midgut, and hindgut tumors

Thirty-four gastroenteropancreatic (GEP) neuroendocrine tumors were evaluated for expression of epithelial (keratin, carcinoembryonic antigen [CEA] and neuroendocrine (neuron-specific enolase, chromogranin, bombesin) markers, and results were correlated with histologic patterns and histochemical staining. Tumors of mixed pattern (insular or trabecular with glandular areas) predominated. CEA localization corresponded to staining for mucin, with polarized apical or lumenal staining in glandular areas. Four trabecular midgut carcinoids, however, revealed diffuse cytoplasmic staining for CEA. Staining for keratin proteins was present in 68% of tumors. Bombesin immunoreactivity was demonstrated in 60% of GEP neuroendocrine tumors, indicating that bombesin positive metastatic tumors may not be predominantly of pulmonary origin, as previously suggested. Chromogranin was a sensitive marker for identifying normal gastrointestinal neuroendocrine cells that were not demonstrated by staining for neuron-specific enolase. Chromogranin was present in most neuroendocrine tumors, but was absent from three of five rectal carcinoids in keeping with the distinctive profile of hormonal and silver staining in these tumors. All GEP neuroendocrine neoplasms expressed both neuroendocrine and epithelial markers, supporting their derivation from endodermal epithelium.     

Expression of pan-neuroendocrine proteins in 53 neuroblastic tumors. An immunohistochemical study with neuron-specific enolase, chromogranin, and synaptophysin

The presence and distribution of pan-neuroendocrine markers such as neuron-specific enolase (NSE), chromogranin (CG), and synaptophysin (SYP) were investigated by immunohistochemistry in 53 cases of neuroblastic tumors, including three cases of ganglioneuromas, 17 ganglioneuroblastomas, and 33 neuroblastomas. In ganglioneuromas, all three markers were observed both in ganglion cells and in neurofibrils. All cases of ganglioneuroblastoma were positive for these markers, however, some variability of staining intensity was noted. Of the 33 cases of neuroblastomas, all were positive for NSE, 23 (70%) for CG, and 31 (94%) for SYP. Neuron-specific enolase was detected not only in the majority of the neuroblasts showing signs of differentiation, but also in some undifferentiated neuroblasts. Chromogranin was found mainly in differentiated neuroblasts with enlarged cytoplasm and nuclei, but was scarcely found in undifferentiated cells. Synaptophysin was detected in some undifferentiated neuroblasts, as well as in differentiated neuroblasts. Two cases without SYP-positive cells were also negative for CG. Our observations conclude that antibodies against NSE and SYP are helpful as a diagnostic aid for neuroblastic tumors.     

A monoclonal antibody against neuron-specific enolase. Immunohistochemical comparison with a polyclonal antiserum

There is skepticism about the value of antisera to neuron-specific enolase (NSE) for immunohistochemical identification of neural and neuroendocrine differentiation in neoplasms, because of reports of detection of NSE, in a large percentage of nonneuroendocrine neoplasms. By immunohistochemical methods, the authors compared a monoclonal antibody to NSE (Mab NSE) with a heterologous antiserum to NSE (Het NSE) on 348 samples of tumors of diverse histogenesis. They studied 93 neural and neuroendocrine tumors and 255 nonneuroendocrine, nonneural tumors. The Mab NSE was slightly less sensitive but clearly more specific than the Het NSE in recognizing neural and neuroendocrine differentiation. Only 2% of the nonneuroendocrine, nonneural tumors reacted positively with Mab NSE; in contrast, 20% of the same tumors were positive with the Het NSE. Moreover, intense nonspecific staining was frequent with Het NSE, which often rendered interpretation difficult. Because of its superior specificity, the Mab NSE used in this study is more valuable than the heterologous antiserum as a diagnostic reagent in tumor diagnosis.     

Small cell neuroendocrine carcinoma of the colon and rectum: clinical, histologic, and ultrastructural study and immunohistochemical comparison with cloacogenic carcinoma

In an effort to provide immunocytochemical data that would be useful in distinguishing between small cell epithelial tumors of the anorectal region, 10 cases of neuroendocrine small cell colorectal carcinoma (NSCCC) and five cases of cloacogenic carcinoma (CC) were studied with antibodies to cytokeratin, epithelial membrane antigen (EMA), chromogranin, blood group isoantigens (BGI), carcinoembryonic antigen (CEA), Leu-M1, Leu-7, leukocyte common antigen (LCA), S-100 protein, neurofilaments (NF), neuron-specific enolase (NSE), serotonin, and 14 neuropeptides. The diagnoses for all 15 tumors were verified ultrastructurally. Among the antigenic determinants considered, reactivity for low- and medium-molecular-weight cytokeratin, EMA, NSE, and NF was seen in the majority of NSCCCs, whereas the CCs were positive for all cytokeratin classes, BGI, EMA, and CEA. In addition, Leu-M1, Leu-7, and chromogranin were each expressed in three cases of NSCCC. None of the other antisera yielded positive results in tumors of either type. All 10 patients with NSCCC died of their tumors within 11 months of clinical presentation, while four of the five CCs proved fatal, with an average survival of 28 months. One of the patients with CC was free of disease 31 months after diagnosis. These data suggest that an immunocytochemical panel, consisting of antibodies to high-molecular-weight cytokeratin, BGI, CEA, NSE, and NF (and possibly Leu-7 and chromogranin as well), is capable of distinguishing between NSCCC and CC in problematic cases. Although tumors of both types are aggressive, it is possible that the survival statistics for both may be improved through more accurate diagnostic classification.     

Paragangliomas of the head and neck: immunohistochemical neuroendocrine and intermediate filament typing

Twenty-nine paragangliomas of the head and neck region including 20 glomus jugulare (GJ) and nine carotid body (CB) tumors were evaluated for the presence of neuroendocrine and intermediate filament antigens. Immunohistochemistry on formalin-fixed, paraffin-embedded tissue was used to identify: S-100 protein (S-100); neuron-specific enolase (NSE); chromogranin A (CHA); serotonin (SER); synaptophysin (SYN); cytokeratin (CK); neurofilament (NF); desmin (DES); vimentin (VIM); and glial fibrillary acidic protein (GFAP). S-100 protein staining of sustentacular cell nuclei and cytoplasm was found in all tumors and was present in chief cells in 4 of 20 GJ and 3 of 9 CB tumors. All tumors stained with at least three neuroendocrine markers (29 of 29 NSE, 28 of 29 SYN, 26 of 29 CHA, 25 of 29 SER). CK was detected in 2 GJ and 1 CB tumor using anticytokeratins AE 1/3 and CAM 5.2. Neurofilament protein could not be demonstrated in fixed material, and all tumors were negative for GFAP and desmin. Vimentin was inconsistently detected in chief and sustentacular cells. We conclude that, in formalin-fixed material, paragangliomas have S-100 protein staining of sustentacular cells with chief cells containing antigens associated with neuroendocrine differentiation. The presence of CK in some paragangliomas is consistent with recent tissue culture studies demonstrating immunoblot confirmation of CK in pheochromocytomas and represents a potential source of immunohistologic misinterpretation in diagnosis, unless a panel of markers is utilized.     

Atypical thymoma (WHO B3) with neuroendocrine differentiation: report of a case

We report a case of type-B3 thymoma manifesting neuroendocrine differentiation. The patient was a 42-year-old woman who complained of shoulder pain but had no symptoms of myasthenia gravis or anemia. The tumor was located in the anterior mediastinum and had directly invaded the pericardium and left lung. Histological examination revealed that the tumor was lobulated by bands of fibrous tissue, perivascular spaces were scattered throughout the tumor, and there were a few intraepithelial lymphocytes. The vast majority of lymphocytes in the perivascular spaces and in the lobulated tumor were immunohistochemically positive for TdT, MIC2, and CD1a. The majority of tumor cells were polygonal and medium or large in size. The tumor cells were weakly positive for synaptophysin, chromogranin A, CD56, and NSE. Small nests of small, relatively uniform polygonal cells were observed facing the fibrous bands. These cells resembled the cells of carcinoid tumors and were strongly positive for NSE, synaptophysin, chromogranin A, and CD56. Ultrastructurally, sparse dense-core granules were observed in the cytoplasm of a few tumor cells. This is a unique case of thymoma with neuroendocrine differentiation, and to the best of our knowledge this is the first such case ever reported.     

BRAF gene mutations are rare events in gastroenteropancreatic neuroendocrine tumors

The BRAF gene, one of the human isoforms of RAF, is activated by ras, leading to cooperative effects in cells responsive to growth factor signals. We studied the frequency of BRAF and k-ras-2 mutations in primary neuroendocrine gastroenteropancreatic (GEP) tumors. Mutation analysis of the BRAF and k-ras-2 genes was performed in 40 primary neuroendocrine tumors of the GEP system. The expression of extracellular signaling-related kinase (ERK) 1/2, an important downstream point of convergence in the ras-RAF-mitogen-activated protein-ERK pathway was analyzed immunohistochemically. We detected one 1796 T-->A BRAF mutation that led to a substitution of valine by glutamic acid at position 599 (V599E) in 40 primary neuroendocrine GEP tumors (3%). We failed to detect specific mutation of the k-ras-2 gene. We identified constitutively activated ERK in almost all neuroendocrine tumor tissues tested irrespective of BRAF mutation or localization or functional activity. These results suggest that BRAF mutations do not have a role in tumorigenesis of neuroendocrine tumors. Nevertheless, activation of the RAF/mitogen-activated protein kinase pathway might have a causative role in the development of neuroendocrine tumors, independent of BRAF or k-ras-2 mutation.     

Distribution of chromogranin A and secretogranin I (chromogranin B) in neuroendocrine cells and tumors.

The distribution of chromogranin A and secretogranin I (chromogranin B) in normal and neoplastic human endocrine tissues was analyzed with two human monoclonal antibodies against chromogranin A, anti-bovine antiserum against chromogranin A, and an anti-rat antiserum against secretogranin I. Western blotting analyses showed both chromogranin A and secretogranin I in normal adrenals, pheochromocytomas, a pituitary adenoma, and in normal pituitary glands, but not in a bladder carcinoma. Rat adrenal medullary and anterior pituitary tissues reacted with the polyclonal chromogranin A and secretogranin I antisera, but not with the two monoclonal chromogranin A antibodies. All antibodies reacted with most of the neuroendocrine cells and tumors examined. Pituitary prolactinomas contained immunoreactive secretogranin I, but not chromogranin A. Analysis of the distribution of chromogranin A and secretogranin I in pancreatic islet cells showed that chromogranin A was found predominantly in the glucagon-producing A cells, whereas secretogranin I was present in less than 5% of islet cells. These results indicate that chromogranin A and secretogranin I are both useful in the characterization of some neuroendocrine cells and neoplasms.     

Expression of erythropoietin and neuroendocrine markers in clear cell renal cell carcinoma

The aim of the study was to investigate the expression of erythropoietin and neuroendocrine markers in clear cell renal cell carcinoma (CCRCC). We retrospectively reviewed the medical records and re‐evaluated histopathological specimens of 33 patients with CCRCC and compared with those of 11 cases of non‐CCRCC. All patients were treated with a partial or radical nephrectomy at St. Olavs Hospital, Trondheim University Hospital, between 2010 and 2016. Thirty‐three patients who were diagnosed with CCRCC had a total of 35 tumours, where 34 of the tumours were CCRCC and one was papillary adenoma. Thirty‐three (97%) of 34 CCRCCs were positive for erythropoietin, and the same 33 (97%) tumours demonstrated strong expression for neuron‐specific enolase (NSE). Two (6%) of 34 CCRCCs had a positive reaction for synaptophysin, and three (9%) of 34 were positive for CD56. Erythropoietin and NSE were negative in non‐CCRCCs, and chromogranin A was negative in all tumours. The above findings suggest that there is a strong association between CCRCC and the expression of erythropoietin and NSE.     

Increased chromogranin A and neuron-specific enolase in rats with chronic nonbacterial prostatitis induced by 17-beta estradiol combined with castration

Although chronic nonbacterial prostatitis (CNBP) is a common diagnosis in middle-aged men, the etiology of this disease remains poorly understood. Neuroendocrine cells play an important role in the neuroendocrine regulation of the prostate, and chromogranin A (CgA) and neuron-specific enolase (NSE) are regarded as classic markers of neuroendocrine cells. This study aimed to determine CgA and NSE levels in a CNBP rat model to evaluate the role of neuroendocrine cells in the pathogenesis of CNBP. For developing a CNBP rat model, we examined the ability of 17-beta estradiol and surgical castration alone or in combination to induce CNBP. Histologic inflammation of the prostate was assessed in CNBP-induced rats by hematoxylin-eosin staining, whereas CgA and NSE protein levels were assessed by immunohistochemistry, Western blot analysis, and enzyme-linked immunosorbent assays. Our results showed that 17-beta estradiol combined with castration successfully induced CNBP and that CgA and NSE levels were increased in the prostate of CNBP rats as compared to those without CNBP. These findings indicate that the neuroendocrine regulation mediated by neuroendocrine cells may be involved in the pathogenesis of CNBP.