RSUME促进小鼠垂体腺瘤细胞AtT-20凋亡

目的探讨小泛素化修饰增强子(RSUME)、核转录因子-κB抑制因子-α(IκB-α)及核转录因子-κB 1(NF-κB1)在小鼠垂体腺瘤中表达及其细胞凋亡的影响。方法用RSUME小干扰RNA(siRNA)转染At T-20细胞后,用蛋白印迹及实时荧光定量PCR检测RSUME、NF-κB1和IκB-α蛋白的表达及mRNA的表达,流式细胞计量术检测细胞凋亡。结果在蛋白水平,RSUME-siRNA转染后RSUME及IκB-α表达下调(P0.05),NF-κB1则上调(P0.05);在mRNA水平,转染后RSUME mRNA表达水平明显低于转染前(P0.05),而IκB-α及NF-κB1 mRNA水平无明显变化;转染后细胞凋亡率明显升高。结论 RSUME可以通过NF-κB1促进小鼠At T-20垂体腺瘤细胞的凋亡。     

TFF3 knockout in human pituitary adenoma cell HP75 facilitates cell apoptosis via mitochondrial pathway

Trefoil factor 3 (TFF3), a regulatory protein composed of 59 amino acids, has been suggested to be involved in pathogenesis, proliferation, differentiation, invasion, migration and apoptosis in multiple malignant tumors. This study thus investigated the effect of TFF3 knockout in human pituitary adenoma cell line HP75 on cell apoptosis and related pathways. RNA interference approach was used to knock down the expression of TFF3 protein. The gene silencing was validated by RNA denaturing gel electrophoresis and Western blotting. The effect of TFF3 knockout on cell apoptosis was analyzed by Western blotting and flow cytometry. TFF3 protein level in pituitary adenoma was about 3.61 ± 0.48 folds of that in normal tissues (P < 0.01). After transfecting with small interference RNA (siRNA) against TFF3, the apoptotic ration was significantly elevated (P < 0.01). Apoptosis related protein Bcl-2 and caspase-3 levels were remarkably depressed after siRNA transfection, while Bax and cleaved caspase-3 levels were elevated. TFF3 protein knockout can facilitate apoptosis of human pituitary adenoma HP75 cells via mitochondrial pathway.     

垂体腺瘤手术入路的解剖学观察

目的为垂体腺瘤的显微外科手术提供解剖学参考依据。方法采用30具成人尸体头颅,按经蝶入路、经额入路、经翼点入路三种手术入路方式逐层解剖入路的邻近血管、神经并进行解剖学观测。结果颈内动脉内口间距(21.73±3.72)mm、视神经管内口间距(11.77±3.34)mm、海绵窦间距(20.92±4.58)mm、鞍底横径(13.57±4.21)mm、鞍底纵径(13.61±3.18)mm、蝶窦前壁至鼻孔的深度(92.91±17.81)mm、眉弓上缘中点到鞍结节的距离(62.24±14.17)mm、眉上缘中点到同侧视神经管内口距离(53.45±16.91)mm、眉上缘中点到对侧视神经管内口距离(62.24±20.80)mm、眉弓上缘中点到同侧颈内动脉入口的距离(69.81±21.96)mm、眉弓上缘中点到对侧颈内动脉入口的距离(78.40±27.46)mm、眉弓上缘中点到同侧颈内动脉膝部的距离(56.43±15.31)mm、眉弓上缘中点到对侧颈内动脉膝部的距离(64.53±17.01)mm、翼点到垂体柄末端距离(59.24±17.17)mm、翼点到前床突的距离(45.51±10.55)mm、翼点到海绵窦的距离(43.72±9.48)mm。结论应用这些解剖学测量结果可指导与鞍区相关的手术,安全显露鞍区解剖结构,不损伤重要的血管和神经。     

经单鼻孔蝶窦入路垂体腺瘤切除45例分析

目的探讨经单鼻孔蝶窦入路显微手术垂体腺瘤切除的方法和效果。方法对45例垂体腺瘤患者采用经单鼻孔蝶窦入路显微手术切除6例微腺瘤及39例大腺瘤,其中泌乳素(PRL)腺瘤18例,生长激素(GH)腺瘤8例,混合性(PRL GH或PRL ACTH)腺瘤12例,无功能性腺瘤7例。结果显微镜下全切除43例,次全切除2例。术后一过性尿崩症28例,经治疗1周左右恢复。无手术死亡、脑脊液鼻漏、术区出血及脑膜炎等并发症。术后平均住院8 d,全组病例术中未输血;随访1~5年,1例复发。结论单鼻孔经蝶窦入路较传统经蝶入路具有安全、简便、微创、术后并发症少、恢复快、效果满意等优点,是鞍内肿瘤切除的良好径路。     

Unusual double pituitary adenoma: a case report

We report the case of a 60-year-old woman with Cushing disease. Magnetic resonance imaging (MRI) revealed a large sellar and suprasellar mass involving the right cavernous sinus, consistent with pituitary macroadenoma. It was resected by transsphenoidal surgery. Light microscopy revealed two separate pituitary adenomas with different histologic and immunohistochemical features. One was amphophilic and strongly Periodic Acid-Schiff (PAS) positive, the other chromophobic and PAS negative. The former tumor was immunopositive for adrenocorticotropic hormone (ACTH); approximately 30% tumor cells were immunopositive for MGMT (O6-Methylguanine-DNA Methyl-Transferase). The second tumor was a PAS negative, luteinizing hormone (LH) and alpha subunit immunopositive gonadotroph adenoma. In this tumor, about 90% of the cells were immunopositive for MGMT. The Ki-67 nuclear indices of the two tumors were 6% and 2%. Our case represents a rare combination of two morphologically different pituitary adenomas, one producing ACTH and the other LH and alpha subunit. The two tumors differed not only in Ki-67 labeling indices but in MGMT immunoexpression as well.     

蝶窦垂体腺瘤的临床病理学观察

目的 探讨蝶窦垂体腺瘤(sphenoid sinus pituitary adenoma,SSPA)的临床病理特征,提高对蝶窦垂体腺瘤的认识和病理诊断水平.方法 对12例SSPA进行临床、组织病理学和SP法免疫组化染色(PRL、hGH、ACTH、LH、TSH、FSH等).结果 12例SSPA,男5例,女7例,年龄39～66岁,平均48.4岁,中位年龄55.5岁.其中原发于蝶窦和原发于垂体窝侵至蝶窦者各6例,原发于蝶窦者年龄较原发于垂体窝者偏大.临床症状多样.镜下,原发于蝶窦的垂体腺瘤的形态学与蝶鞍垂体腺瘤相同;SSPA可伴有较明显的慢性炎症细胞浸润(2/12,16.67%).10例行免疫组织化学染色的结果,瘤细胞PRL阳性6例;瘤细胞各项标记物均阴性2例;瘤细胞ACTH阳性、hGH阳性各1例.结论原发于蝶窦的垂体腺瘤比较少见,主要根据光镜及免疫组化确诊,应注意与慢性蝶窦炎和浆细胞瘤鉴别.     

Role of sex hormones in development of pituitary adenoma

Role of sex hormones in the development of pituitary adenomas was investigated by analyzing the content of nuclear estradiol and testosterone receptors in different tumors of the anterior pituitary: prolactinomas, meningiomas, growth hormone-producing adenomas, astrocytomas, neurinomas, and ependymomas. The concentration of nuclear estrogen and androgen receptors in prolactin-secreting pituitary adenomas was much higher than in growth hormone-producing adenomas and other pituitary tumors.     

RGD-FasL基因的构建、表达、纯化及其活性分析

目的构建适于原核表达的重组蛋白RGD-FasL表达载体,并进行重组蛋白的表达纯化及抗肿瘤活性分析。方法通过重叠PCR将RGD序列插入到FasL基因的N端,获得RGD-FasL基因,构建pGEX-5X-1/RGD-FasL表达载体。转化大肠杆菌BL21(DE3),IPTG诱导表达,GST柱纯化。采用体外黏附实验、MTT比色法、流式细胞法检测融合蛋白的功能。结果通过重叠PCR获得了编码正确氨基酸序列的目的基因。目的蛋白以分泌的形式表达,表达量占菌体总蛋白的30%以上。纯化后,蛋白纯度达95%以上。体外黏附实验表明所纯化的融合蛋白可与宫颈癌Hela细胞发生特异结合。MTT比色法与流式细胞技术均表明纯化的融合蛋白能特异性地诱导肿瘤细胞发生凋亡。结论重组蛋白RGD-FasL表达载体的成功构建、表达、纯化及活性分析,为进一步的功能研究奠定了基础。     

垂体瘤复发因素分析及治疗

目的探讨复发性垂体瘤二次手术的效果及并发症。方法回顾分析本院10年间12例复发性垂体瘤的手术,肿瘤大小(2.3cm×2.3cm)~(6.0cm×4.5cm),复发的肿瘤大多为经蝶术后。结果9例肿瘤获全切,3例次全切除。术后2例复发行第3次手术,其余未见复发。结论初步研究结果显示,复发原因多样,复发性垂体瘤应首选手术,全切率高。肿瘤的位置是术式选择的关键,不推荐保守治疗。     

甘丙肽抑制大鼠垂体腺瘤细胞体外侵袭性

目的 探讨甘丙肽对大鼠垂体腺瘤细胞侵袭性的作用及其受体机制.方法 提取大鼠垂体腺瘤GH3细胞RNA,反转录后测定甘丙肽及其3个受体亚型的表达情况;将大鼠垂体腺瘤GH3细胞分为对照组、甘丙肽药物处理组和选择性甘丙肽2型受体激动剂AR-M1896组,利用MTT方法检测对照组和实验组在给药后12、24和36 h各分组细胞活力...     

IL-2对培养的人垂体腺瘤细胞增殖的影响及其机制

目的:研究IL-2对人垂体腺瘤细胞的增殖、细胞周期、细胞内蛋白激酶C(PKC)及cAMP/cGMP的影响,并探讨其作用的机制。方法:采用MTT比色法和3H-TdR掺入法检测IL-2对人垂体腺瘤细胞增殖和DNA合成的影响;用流式细胞术检测IL-2对垂体腺瘤细胞细胞周期的影响。用放射免疫测定法检测PKC的活性及cAMP和cGMP的含量。结果:(1)IL-2可促进垂体腺瘤细胞增殖和DNA合成,且呈剂量依赖性。(2)IL-2可显著降低垂体腺瘤细胞G1期的细胞比率,而增加S期和G2期的细胞比率。(3)与空白处理组相比较,使用PKC的激动剂PMA处理培养的人垂体腺瘤细胞,可使胞膜和细胞总PKC升高;而用IL-2(1×105U/L)处理后,胞质、胞膜和细胞总PKC的活性均升高。(4)IL-2(5×104、1×105U/L)作用于人垂体腺瘤细胞后,胞内cAMP浓度显著降低;而cGMP的浓度没有明显改变。结论:IL-2对垂体腺瘤细胞增殖分化的调控作用是细胞内多信息系统相互整合的结果。     

青蒿琥酯诱导垂体泌乳素瘤细胞凋亡并下调泌乳素蛋白表达

目的 初步研究青蒿琥酯( artesunate,ART)对泌乳素瘤细胞及其泌乳素(prolactin,PRL)蛋白表达的作用效果.方法 MTT观察青蒿琥酯作用细胞后,细胞增殖情况,并计算药物的半数抑制浓度(IC50),Hoechst33342染色观察细胞凋亡小体,TUNEL计算细胞凋亡率,Western blot和RT...     

Prolactin-producing pituitary adenoma associated with prolactin cell hyperplasia

A 24-yr-old woman with amenorrhea, galactorrhea, hyperprolactinemia, and sellar mass underwent transsphenoidal surgery. Histologic, immunohistochemical, and electron microscopic investigation revealed a well-differentiated, sparsely granulated prolactin (PRL) cell adenoma of the pituitary showing conclusive PRL immunoreactivity. In the nontumorous adenohypophysis PRL cell hyperplasia was noted. Marked differences were evident between the neoplastic and hyperplastic areas. The tumor consisted of sparsely granulated PRL cells immunoreactive only for PRL. As demonstrated by immunoelectron microscopy, the hyperplastic are a comprised monohormonal sparsely granulated PRL cells as well as bihormonal mammosomatotrophs immunoreactive for both PRL and growth hormone. The MIB-1 index was higher whereas microvessel density was lower in the adenoma as compared with the hyperplastic area. In addition, the nontumorous area showed lymphocytic infiltration whereas inflammatory reaction was not seen in the adenoma. This case represents a rare association of a PRL cell adenoma and PRL cell hyperplasia. The fact that these two lesions were contiguous in the surgically removed material raises the possibility that hyperplasia can precede and transform into adenoma.     

眶上锁孔入路治疗垂体瘤的临床解剖学研究

目的 :探讨内窥镜辅助眶上锁孔入路治疗垂体瘤的可行性。方法 :2 1例福尔马林固定尸体头部标本用于鞍区各解剖结构 ,特别是垂体柄、视神经、视交叉及其供血动脉特点的观察 ,总结手术可利用的间隙、应保护的结构 ;在 9例新鲜尸头上模拟进行内窥镜辅助眶上锁孔入路手术 ,进一步验证其可行性及优势。结果 :颈内动脉床突上段长度 (14 .5± 1.3 )mm(8.1～ 18.5mm ) ,发向垂体柄、视神经或视交叉的穿支动脉的支数分别为 :大脑前或前交通动脉 3 .0支 (2～ 6支 ) ,颈内动脉 2 .1支 (1～ 5支 ) ,后交通动脉 3 .2支 (3～ 6支 ) ,基底动脉 1.4支 (1～ 3支 )。视神经颅内段长度为 (11.4± 2 .7)mm (6.1～ 17.6mm ) ,第 1间隙面积为 (4 4 .8± 3 .4)mm2 (7.0～ 10 0 .8mm2 ) ,手术可通过第 1间隙或 /和第 2间隙进行。结论 :通过眶上锁孔入路治疗向鞍上发展的垂体瘤有充足的操作空间 ,具有视神经、视交叉减压充分 ,利于保护其供血动脉的优点。     

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。     

经颅与经蝶窦分期手术切除巨大垂体腺瘤

目的 探讨经颅入路与经蝶窭入路分期切除不同生长方式的巨大垂体腺瘤的手术方法及临床意义。方法 回顾性分析我科行经颅与经蝶窭分期手术的12例巨大垂体腺瘤患者的临床表现、影像学特点，根据肿瘤的不同形态及生长方式，结合手术入路和手术方式探讨其手术疗效。结果 本组病例根据生长方式的不同分为：①单纯向上呈葫芦形生长，经额下入路及经蝶窭入路手术者4例；②向鞍上及鞍旁生长，行经翼点入路及蝶窭入路手术者6例；③向三脑室及侧脑室生长，经侧脑室入路及经蝶窭入路手术者2例。二次手术间隔时间1～3月间不等，均采用显微手术治疗，其中肿瘤全切除8例，次全切除3例，大部切除1例，无死亡发生。结论 手术前应根据病史和影像学资料判断肿瘤的形态、生长方式、所累及的解剖腔隙和重要结构，选择合理的分期手术方法，才能取得较好的临床治疗效果。     

ACTH-producing pituitary adenoma in an infant with cysts of the kidneys and lungs

The light microscopical, electron microscopical and immunohistological features of a rare ACTH-producing chromophobe adenoma of the pituitary gland in an 11-month-old infant are presented. An unusual histological feature was the presence of numerous follicles. These follicles may develop secondary to necrosis of individual tumour cells. There is only one previous report in the literature of a pituitary adenoma occurring in infancy and that tumour also produced ACTH and showed a follicular histological pattern. It is of interest that our patient also had a combination of medullary cysts of the kidneys and congenital peripheral cysts of the lungs. The association of cysts in lungs with cysts in kidneys also has been reported only once previously. We conclude that the occurrence of cysts in kidneys and lungs and a pituitary tumour in infancy represents a bizarre coincidence although an unknown common embryonic insult to these organs cannot be ruled out.     

Characterization of aryl hydrocarbon receptor interacting protein (AIP) mutations in familial isolated pituitary adenoma families

Familial isolated pituitary adenoma (FIPA) is an autosomal dominant condition with variable genetic background and incomplete penetrance. Germline mutations of the aryl hydrocarbon receptor interacting protein (AIP) gene have been reported in 15–40% of FIPA patients. Limited data are available on the functional consequences of the mutations or regarding the regulation of the AIP gene. We describe a large cohort of FIPA families and characterize missense and silent mutations using minigene constructs, luciferase and β‐galactosidase assays, as well as in silico predictions. Patients with AIP mutations had a lower mean age at diagnosis (23.6±11.2 years) than AIP mutation‐negative patients (40.4±14.5 years). A promoter mutation showed reduced in vitro activity corresponding to lower mRNA expression in patient samples. Stimulation of the protein kinase A‐pathway positively regulates the AIP promoter. Silent mutations led to abnormal splicing resulting in truncated protein or reduced AIP expression. A two‐hybrid assay of protein–protein interaction of all missense variants showed variable disruption of AIP‐phosphodiesterase‐4A5 binding. In summary, exonic, promoter, splice‐site, and large deletion mutations in AIP are implicated in 31% of families in our FIPA cohort. Functional characterization of AIP changes is important to identify the functional impact of gene sequence variants. Hum Mutat 31:1–11, 2010. © 2010 Wiley‐Liss, Inc.