垂体腺瘤中垂激素与S—100蛋白免疫组化双重染 …

目的：探讨滤泡星状细胞在垂体腺瘤分类中的意义及滤泡星状细胞与内分泌细胞之间的关系。方法：应用免疫组化双重染色方法,对４２例人重体腺瘤的垂体激素与Ｓ－１００蛋白表达进行对照观察。结果：垂体腺瘤组织中的滤泡星状细胞有两种情况,一种为腺瘤组织中可见散在分布的滤泡星状细胞,并可见１个瘤细胞既有Ｓ－１００蛋白表达,又含激素分泌颗粒;另一种为滤泡星状细胞构成了腺瘤的一种主要的细胞成分。结论：滤泡星状细胞与内分     

垂体腺瘤中E-cadherin和nm23蛋白表达与肿瘤生物学行为的关系

目的探讨垂体腺瘤中Ecadherin(Ecad)和nm23蛋白表达与肿瘤体积、激素分泌和侵袭性的关系。方法应用免疫组化SP法检测Ecad和nm23基因蛋白在23例侵袭性垂体腺瘤和24例非侵袭性垂体腺瘤组织中的表达。结果Ecad和nm23的表达在侵袭性垂体腺瘤组低于非侵袭性组(P<0.05),Ecad表达在非分泌型垂体腺瘤组低于分泌型组且与肿瘤体积呈负相关(P<0.05),Ecad与nm23表达间呈正相关(P<0.05)。结论Ecad与nm23表达降低可能与垂体腺瘤的侵袭性有关,Ecad表达降低可能影响垂体腺瘤细胞分化和促进细胞增殖,Ecad和nm23可作为评估垂体腺瘤侵袭能力的生物学指标之一。     

P16蛋白表达与垂体腺瘤侵袭和复发的关系

目的 探讨Ｐ１６蛋白表达与垂体腺瘤侵袭及复发的关系。方法 应用免疫组化染色技术检测５７例垂体腺瘤中Ｐ１６蛋白的表达水平,并对患者进行了５抻随访观察。结果 垂体腺瘤细胞中Ｐ１６蛋白表达的阳性率为３１．６％;侵袭性组中该蛋白表达的阳性率显著低于非侵袭性组（Ｐ＜０．０１）。复发组１１例未见该蛋白表达。结论 Ｐ１６蛋白的表达阳性率与垂体腺瘤激素分泌功能无明显的相关性,但其表达水平可作为临床评价垂体腺瘤的侵袭性和预测患者复发的指标。     

垂体腺瘤细胞培养的实验研究

目的 通过对垂体腺瘤细胞体外培养以垂体腺瘤细胞为移植物供体进行颅外移植,补充垂体功能不足及为腺垂体移植细胞库的建立奠定基础。方法 利用术中切除的垂体腺瘤组织进行了体外培养,并通过染色和放射免疫测定技术,评价体外培养的垂体腺瘤组织的功能状态,寻找其功能最佳间期的规律。结果 没有NGF和HRP介入的情况下,垂体腺瘤细胞的形态和功能状态在培养的第6天为最佳。结论 体外培养的垂体腺瘤细胞仍具有分泌激素的功能,为以垂体腺瘤细胞为移植物供体进行颅外移植的研究奠定了实验基础。     

混合性节细胞瘤/垂体腺瘤4例临床病理观察

目的探讨混合性节细胞瘤／垂体腺瘤发生机制、临床病理特征、诊断及鉴别诊断。方法复习4例混合性节细胞瘤／垂体腺瘤患者的临床资料，并对其进行组织学观察和免疫组化（EnVision—plus法）标记。结果4例混合性节细胞瘤／垂体腺瘤患者均为女性，年龄10～35岁，平均26．5岁。临床上3例表现内分泌症状，1例有癫痫症状。组织学上肿瘤由两种不同成分组成，一种结构主要由较多神经节细胞组成，节细胞体积大，可见圆形的尼氏小体，核大，核仁突出，免疫组化标记瘤细胞表达CgA、Syn、S-100蛋白，不表达GFAP；另一部分腺瘤细胞大小形态较一致，圆形或卵圆形，胞质丰富，嗜酸性或空淡，间质血窦丰富，瘤细胞表达GH和PRL。结论混合性节细胞瘤／垂体腺瘤是蝶鞍区极少见的肿瘤，好发于青年女性，常合并肢端肥大症。该瘤的诊断主要依靠组织病理学，并辅以免疫组化标记。治疗主要采用单纯手术切除，预后较好。     

神经生长因子受体在垂体腺瘤中的表达

目的：探讨两种神经生长因子（ＮＧＦ）受体（ＴｒＫＡ和ｇｐ７５）在各类垂体腺瘤中的表达。方法：采用免疫细胞化学ＳＡＢＣ法,检测了４４例经病理证实的垂体腺瘤中两种ＮＧＦ受体的表达情况。结果：ＴｒＫＡ在垂体ＰＲＬ分沁腺瘤中的过度表达率６９．２％,而在其它类型垂体腺瘤中的过度表达率为１９．４％,两者比较差异有非常显著性（Ｐ〈０．００５）。ｇｐ７５在垂体ＰＲＬ分泌腺瘤中的过度表达率在４６．２％,而在其它类型     

垂体腺瘤细胞培养的实验研究

目的通过对垂体腺瘤细胞体外培养以垂体腺瘤细胞为移植物供体进行颅外移植,补充垂体功能不足及为腺垂体移植细胞库的建立奠定基础. 方法利用术中切除的垂体腺瘤组织进行了体外培养,并通过染色和放射免疫测定技术,评价体外培养的垂体腺瘤组织的功能状态,寻找其功能最佳间期的规律. 结果没有NGF和HRP介入的情况下,垂体腺瘤细胞的形态和功能状态在培养的第6天为最佳. 结论体外培养的垂体腺瘤细胞仍具有分泌激素的功能,为以垂体腺瘤细胞为移植物供体进行颅外移植的研究奠定了实验基础.     

PTTG基因蛋白的表达与垂体腺瘤侵袭性和增殖能力的关系

目的探讨人垂体腺瘤中垂体肿瘤转化基因(PTTG)蛋白的表达与肿瘤侵袭性和增殖程度的关系。方法采用免疫组织化学染色方法检测手术切除石蜡包埋的63例垂体腺瘤(侵袭组45例,非侵袭组18例)组织中PTTG蛋白的表达,染色增殖细胞核抗原(PCNA),同时计数组织内微血管数量(MVD)。结果 PTTG在侵袭性垂体腺瘤中的表达水平显著高于非侵袭性垂体腺瘤。侵袭性垂体腺瘤中PCNA标记指数和微血管密度也显著高于非侵袭性垂体腺瘤。相关分析显示PTTG表达与垂体腺瘤内PCNA标记指数和微血管密度呈正相关(P<0.05)。结论 PTTG在垂体腺瘤形成过程中起重要作用,并与垂体腺瘤的侵袭性和增殖程度密切相关。     

兔垂体干细胞的培养鉴定及分化

背景：脑垂体是人体重要的内分泌器官。多种疾病可引起脑垂体损伤，如脑垂体瘤、垂体功能减退症等。垂体干细胞因具有自我更新和多向分化潜力的特性，有望成为一种新的治疗方式修复受损的垂体。目的：采用悬浮细胞球培养法分离培养垂体干细胞，鉴定其增殖及分化能力。方法：采用悬浮细胞球培养法从新生新西兰白兔垂体中分离培养垂体干细胞，观察其形态特征；免疫荧光细胞染色检测垂体干细胞标志蛋白SOX2及Nestin的表达；采用EdU标记法检测垂体干细胞增殖能力；经贴壁及诱导分化后，通过ELISA法检测培养基中的激素水平。结果与结论：通过悬浮细胞球培养法可成功分离得到垂体干细胞球，且具有较强的增殖能力；干细胞特异性标记物SOX2和Nestin阳性表达；经诱导分化后培养基中促肾上腺皮质激素、促甲状腺激素、促生长激素、促黄体生成激素、促卵泡素、催乳素水平显著升高(P <0.001)，具有较强的分化能力。     

CgA、CgB 和 SgⅡ在垂体腺瘤中的表达

本文应用ＣｇＡ、ＣｇＢ和ＳｇⅡ的３种抗体，通过ＳＡＢＣ方法，对８４例按功能分类的垂体腺瘤进行了研究。实验结果表明，ＣｇＡ、ＣｇＢ和ＳｇⅡ表达的阳性率分别为２５．０％、９５．２％和９１．７％。ＣｇＢｔ和ＳｇⅡ两者的表达无明显差异；而与ＣｇＡ相比，则有极显著的差异。从而证明，垂体腺瘤细胞中的神经内分泌颗粒有不同的类型、不同的分布。应用ＣｇＡ、ＣｇＢ和ＳｇⅡ同时标记垂体腺瘤，对该肿瘤的诊断及分型有一定的意义。     

正常人腺垂体和垂体腺瘤中滤泡星状细胞的免疫组织化学研究

Significance of the appearance of folliculo-stellate cells (FSC) was studied in 59 human adenohypophyses and 58 pituitary adenomas after being stained with anti-S100 protein and 6 anti-pituitary hormone antibodies. S100 protein positive cells, stellate in shape with expending cytoplasmic processes among endocrine cells (EC) appeared in all the human adenohypophyses and had a tendency to be clustered in small groups characterized by gathering of 3 to 5 cells in the alveoli. Age or sex difference seemed to have no influence on the distributive density of FSC. FSC in the pituitary adenomas may be derived from two origins. One was the residue of normal pituitary tissue left in adenomas, and the other one seemed to be the chief component of the tumor, known as folliculo-stellate cell adenoma.     

S-100 protein immunopositivity in human nontumorous hypophyses and pituitary adenomas

The presence, distribution, and morphological appearance of S-100 protein-immunoreactive cells in the human hypophysis were studied by immunocytochemistry. One hundred and twelve nonadenomatous pituitaries from fetuses to adults and pituitaries affected by several lesions including metastases, acute infarcts, and lymphocytic hypophysitis, as well as 115 pituitary adenomas were examined.S-100 protein immunoreactivity was detected in neurohy-pophyseal pituicytes and stellate cells of the pars distalis from 5 months following birth. In adults, S-100 protein-immunopositive cells displayed a preferential topographical association with growth hormone-, follicle-stimulating hormone-, luteinizing hormone-, and alpha-sub-unit-immunoreactive cells and with capillary walls. Colloid-containing follicles were mainly lined by hormone-containing cells, although scattered S-100 protein-immunoreactive processes or cell bodies were also observed forming their walls.No major changes in S-100 protein-immunoreactive cells were observed in the pituitary parenchyma bordering metastatic, inflammatory, necrotic, or adenomatous tissues. Eighteen of 115 pituitary adenomas contained a variable number of S-100 protein-immunoreactive cells. No preferential association of these cells with any type of pituitary adenoma was found.We propose that S-100 protein expression in the nontumorous adenohypophysis and pituitary adenomas may constitute a dynamic process and that S-100 protein-positive cells may constitute a heterogeneous cell population composed of pure, fully differentiated stellate cells and of transdifferentiated follicular cells.     

Plurihormonal pituitary adenomas: immunostaining of all pituitary hormones is mandatory for correct classification

AIMS: We studied the clinicopathological characteristics of plurihormonal pituitary adenomas. METHODS AND RESULTS: The study material included 167 plurihormonal adenomas, which consisted of 31% of the surgically removed pituitary adenomas that we collected during a 12-year period. The mean age of patients with plurihormonal adenoma was 45.7 years (range 13-75 years). There were 86 men and 81 women. All tumours were fully classified by immunohistochemical staining for seven pituitary hormones or subunits. Thirty immunohistochemical subtypes of plurihormonal adenomas were recognized. Hormonal symptoms were present in 70% of patients, while serum hormonal levels were increased in 89% of patients. Most patients had symptoms related to only one of the hormones and only 7% of patients had symptoms related to two hormones. The most common hormonal symptom was acromegaly (50%); symptoms related to hyperprolactinaemia ranked second (20%). Double immunostaining of all the possible combinations of the hormones was performed in 30 selected tumours, and they all showed mixtures of hormones in individual adenoma cells in any hormonal combinations studied. The latter finding supported the view that plurihormonal adenomas are monomorphous adenomas. CONCLUSIONS: Plurihormonal adenomas are common pituitary adenomas. Immunohistochemical staining of all pituitary hormones is mandatory for correct classification.     

Histological changes in the hypofunctional pituitary gland following conventional radiotherapy for adenoma

AIMS: Although delayed hypopituitarism is a common complication of conventional radiotherapy of sellar tumours, histological changes that may account for it have been rarely reported. To elucidate the changes, hypofunctional pituitary glands following irradiation were studied. METHODS AND RESULTS: Two pituitary glands obtained at autopsy from patients who had been irradiated for adenoma and exhibited hypopituitarism were examined. In both cases diffuse fibrosis was observed in the adenohypophysis, whereas the neurohypophysis remained unchanged. Immunohistochemistry showed that stellate-shaped S100 protein-positive cells were increased in number and distributed among the endocrine cells. Some irradiated endocrine cells showed dense granular immunoreactivity for mitochondrial protein, cytochrome oxidase and manganese-superoxide dismutase. In addition to faint reactivity with anti-cytokeratin 8, 18 antibody, many cells were densely positive with anti-cytokeratin 1, 5, 10, 14 antibody. CONCLUSIONS: These results indicate that radiation- induced fibrosis is associated with an increased number of folliculo-stellate cells and the presence of metabolic dysfunctional mitochondria resembling mitochondria in oncocytes. Squamous metaplasia in the irradiated endocrine cells was also noted. Various intracellular changes may participate in delayed pituitary hypofunction following radiotherapy.     

Folliculo-stellate cells in pituitary adenomas of patients with acromegaly

Pituitary adenoma tissue from patients with acromegaly (n = 286) was obtained by surgery and examined for folliculo-stellate cells by immunostaining for S100 protein. The number of immunostained cells varied from one adenoma to another. A hundred and ninety-eight pituitary adenomas (69%) contained S100 protein positive cells corresponding to folliculo-stellate cells (FSC): in 100 cases (35%), only few sparse FSC were found, in 43 cases (15%) FSC were scattered throughout the adenoma, and in 55 cases (19%) there was an abundance of FSC. There were no significant differences regarding sex or age of the patients. The relative amount of FSC vaired among different adenoma types. Plurihormonal adenomas showed the highest FSC density, whereas the majority of monohormonal adenomas contained only few or no FSC. Patients with pituitary adenomas containing scattered FSC had a significantly higher preoperative mean GH level than patients with pituitary adenomas not having FSC. There was a negative correlation between the FSC density in adenoma tissue and the preoperative mean PRL level. There was no correlation between the tumour size and the amount of FSC. Our data indicate no effect of the duration of symptoms on the FSC density and do not suggest a correlation between FSC density and the percentage of a certain hormone-secreting cell type.     

Clinicopathologic features of familial pituitary adenomas

Pituitary adenomas are common neoplasms. Initially considered as sporadic tumours, some of them are associated with familial syndromes such as familial isolated pituitary adenoma, multiple endocrine neoplasia type 1 and type 4, X-linked acrogigantism syndrome, Carney complex, pheochromocytoma/paraganglioma–pituitary adenoma, pituitary blastoma and McCune–Albright syndrome. They represent a group of diseases with different genetic background and variable phenotype. Here, we summarize the clinicopathological features of pituitary adenomas associated with these familial syndromes.     

Glucocorticoid receptor expression in nontumorous human pituitaries and pituitary adenomas

Glucocorticoids have multiple actions, including a suppressive feedback effect on pituitary corticotrophs via the glucocorticoid receptor (GR). By immunocytochemistry, we studied GR expression in 86 surgically removed various pituitary adenoma types. Ten cases contained nontumorous pituitary fragments, which were suitable for immunocytochemical investigation. In addition, 30 autopsy-obtained pituitaries, 10 of them containing incidental microadenomas, were examined as well. Using a polyclonal GR antibody, the streptavidin-biotin-peroxidase complex method revealed nuclear and/or cytoplasmic GR immunoreactivity in many nontumorous corticotrophs and other adenohypophysial cell types and in S-100 protein immunopositive stellate cells. Cellular localization was confirmed by double immunostaining. Pars intermedia corticotrophs, posterior lobe axons, Herring bodies, and pituicytes as well as several endothelial cells lining the capillaries were also immunopositive. GR immunoreactivity was also demonstrated in many GH, PRL, ACTH, TSH, FSH, LH α-subunit producing adenomas, null cell adenomas, and oncocytomas. The extent and degree of immunostaining varied considerably from case to case. Suppressed corticotrophs showing the Crooke’s hyaline change due to glucocorticoid excess were present in the nontumorous pituitaries of patients with Cushing’s disease and in those treated with pharmacologic doses of glucocorticoids. Many suppressed nontumorous corticotrophs exhibited only weak or no GR immunopositivity, indicating GR downregulation accompanied by cellular injury. Study of autopsy obtained pituitaries for GR yielded inconclusive results indicating that autopsy obtained adenohypophyses are not suitable for the immunocytochemical investigation of GR.     

Plurihormonal pituitary adenomas

Plurihormonal adenomas of the pituitary, ie, tumors that engage in the production of unusual combinations of hormones, represent approximately 10% to 15% of all adenomas. Such tumors comprise in excess of 50% of adenomas in the setting of acromegaly and occur with somewhat greater frequency in childhood and adolescence than in adulthood. Eight percent are associated with multiple endocrine neoplasia, type I. The most common variant of plurihormonal adenoma produces growth hormone, prolactin, and one or more glycoprotein hormones, the most common being TSH. Clinical effects most often reflect the presence of growth hormone, and to a lesser extent, prolactin cells; expression of glycoprotein hormone production is rare. The tumors are more often macroadenomas (80%) than microadenomas (20%) and demonstrate gross invasion in 50% of cases. Plurihormonal adenomas may be ultrastructurally monomorphous, bimorphous, or trimorphous; thus, one morphologic cell type may elaborate several hormones.     

Detection of inhibin α and βA subunits (inhibin A, B, activin A,AB) in the human pituitary gland and in pituitary adenomas by immunohistochemistry and nonradioisotopicin situ hybridization

Inhibin and activin are gonadal hormones produced in human ovaries. They are known to act on anterior pituitary cells to regulate the synthesis and secretion of follicle-stimulating hormone (FSH). The purpose of the present study was to determine the localization of inhibin and activin subunits α and βA as endocrine markers in the human normal pituitary gland and pituitary adenomas, using immunohistochemistry andin situ hybridization (ISH) methods. Pituitary tissues from surgical and autopsy materials were fixed in 10% formalin and embedded in paraffin. Five normal pituitary glands and 79 pituitary adenomas were immunostained with the avidin-biotin peroxidase complex (ABC) method using polyclonal antibodies against inhibin and activin subunits α and βA. The other antibodies against anterior pituitary hormones used in this study were as follows: antigrowth hormone (anti-GH), antiprolactin (anti-PRL), antiadrenocorticotropic hormone (anti-ACTH), anti-FSHβ, antilutenizing hormone (anti-LH) β, antithyroid-stimulating hormone (anti-TSH) β, and antiglycoprotein α-subunit (anti-α-SU). We analyzed gene expressions of subunits α and βA by nonradioisotopic ISH in pituitary adenomas. In the normal human pituitary glands, inhibin and activin subunits α and βA immunoreactivities were found diffusely in the cytoplasm of anterior pituitary cells. The percentage of subunit α-immunopositive cells was 40% of the anterior pituitary cells. Subunit βA immunoreactivities were observed in about 15% of the anterior pituitary cells. By the double-staining method, subunit α immunoreactivity was detected in all types of anterior pituitary cells, and it was colocalized most frequently with GH and α-SU-positive cells. Subunit βA immunoreactivity was colocalized predominantly with PRL, FSH-β, LH-β, and α-SU. Among the 79 adenomas, 75 cases (94.9%) were positive for subunit α, and 50 cases (63.3%) were positive for subunit βA. Subunit βA was positive in tumor cells with the following incidences: GH adenomas, 3 of 14 (21.4%); PRL adenomas, 5 of 8 (62.5%); ACTH adenomas, 6 of 6 (100%); TSH adenomas, 7 of 7 (100%); nonfunctioning adenomas, 29 of 44 (65.9%), including gonadotropin-positive, 16 of 22 (80.0%). The ISH signals for subunits α and βA were strongly expressed in gonadotropin-positive adenomas among the nonfunctioning adenomas. The mRNA signals were low and infrequent in the GH-producing adenomas. Inhibin and activin subunit α localization did not demonstrate cell-type specificity in pituitary adenomas. In contrast, subunit βA demonstrated predominant positivity in the functioning pituitary adenomas (ACTH- and TSH-secreting) and nonfunctioning adenomas (including gonadotropin-positive adenomas). The present results suggest that the functional role of inhibin and activin in the differentiation of cells in normal human pituitary glands and adenomas is present in subunit βA.