【摘要】：前言垂體腺瘤是顱內常見的良性腫瘤,約占顱內腫瘤的10%～25%。它們起源于高度分化的分泌一種或多種激素的垂體前葉細胞。根據病理類型,垂體腺瘤可以分為生長激素(growth hormone, GH)腺瘤、泌乳素(prolactin, PRL)腺瘤、促腎上腺皮質激素腺瘤(adrenocorticotrophic hormone, ACTH)腺瘤、無功能(nonfunctioning,NF)腺瘤等,不同類型腫瘤臨床表現亦不同,例如肢端肥大癥、閉經/溢乳、庫欣綜合征或僅僅表現為壓迫周圍結構等癥狀。無論何種病理類型,大多數垂體腺瘤生長緩慢,但少數垂體腺瘤表現為惡性生物學行為,例如侵襲性快速生長,對常規(guī)手術、放射、藥物治療無反應等。因此,判斷腫瘤生物學行為有助于判斷腫瘤預后和治療。因為目前常規(guī)的組織病理學檢查不能有效地區(qū)別不同垂體腺瘤細胞細微的形態(tài)差別和生物學意義。因此,進一步研究垂體腺瘤發(fā)生、發(fā)展的分子生物學機制,發(fā)現腫瘤標志物及其臨床意義,對于垂體腺瘤的預防、診斷、預后評價和治療選擇將有重大的意義。Janus蛋白酪氨酸激酶(Janus protein tyrosine kinase, JAK)/信號轉導和轉錄激活因子(Signal transducer and activator of transcription, STAT)信號通路是一條由細胞外至細胞核的信號轉導極其快速的信號通路。目前在哺乳動物已鑒定出STAT家族的7個成員：STAT1、STAT2、STAT3、STAT4、STAT5a、STAT5b和STAT6。這其中以STAT3研究最為廣泛。STAT3蛋白在細胞質中呈潛伏狀態(tài),被磷酸化后的STAT3即變?yōu)榛罨问降腟TAT3 (p-STAT3),形成同源或異源二聚體,易位至細胞核與特定的啟動子結合,誘導靶基因(如PTTG、VEGF、MMP2、Cyclin-Dl等)表達從而調控細胞增殖、分化、凋亡、血管形成等過程,如果STAT3被激活后持續(xù)活化可導致上述靶基因異常高表達,從而表現出致癌作用。目前,關于垂體腺瘤組織中STAT3表達和激活情況以及與下游靶基因PTTG的關系,國內外尚未見專題報道。目的通過免疫組化、Western blot. Real Time PCR檢測垂體腺瘤組織中STAT3、 p-STAT3. PTTG蛋白和mRNA表達水平,分析這三種基因編碼蛋白在垂體腺瘤發(fā)生發(fā)展中的作用以及STAT3、p-STAT3、PTTG與垂體腺瘤臨床病理因素之間的關系,探討STAT3調節(jié)PTTG的表達是否參與垂體腺瘤的致病過程。材料與方法垂體腺瘤標本來自2010年5月至2013年6月在北京協和醫(yī)院神經外科接受手術治療的80例垂體腺瘤患者。所有手術標本均經臨床及病理診斷證實。腫瘤標本包括20例PRL腺瘤,20例ACTH腺瘤,20例GH腺瘤,20例NF腺瘤。其中,男性27例,女性53例,年齡17-58歲,平均40歲。所有病例均未進行包括化療和放療等術前治療。根據Knosp標準,Ⅲ、Ⅳ級定義為侵襲性垂體腺瘤,Ⅰ、Ⅱ級定義為非侵襲性腺瘤,其中侵襲性腺瘤30例,非侵襲腺瘤共50例。結果1、免疫組化STAT3蛋白細胞核和細胞漿均有著色。STAT3蛋白在GH腺瘤的評分(6.15±2.90)顯著高于PRL腺瘤(3.90±2.29) ACTH腺瘤(3.55±1.53)和NF腺瘤(1.85±1.73),差異具有統計學意義(p0.01)。PRL腺瘤和ACTH腺瘤中STAT3蛋白的評分差異無統計學意義(p=0.57)。NF腺瘤評分最低,與GH、PRL和ACTH腺瘤相比,差異具有統計學意義(p0.05)。上述結果被Western blot結果證實。2、免疫組化P-STAT3蛋白細胞核著色。p-STAT3蛋白在GH腺瘤的免疫組化評分最高(3.20±1.50),與其他三種類型腫瘤相比,差異具有統計學意義(p0.01)。PRL腺瘤(1.50±1.23)和ACTH腺瘤(1.15±1.39)評分次之,NF腺瘤評分最低(0.85±0.74),這三種類型之間比較,差異無統計學意義(p0.05)。上述結果被Western blot結果證實。3、免疫組化PTTG蛋白細胞漿著色。PTTG蛋白在GH腺瘤評分最高(2.05±0.99),與其他三種類型腫瘤相比,差異具有統計學意義(p0.05)。ACTH腺瘤(1.60±1.05)和PRL腺瘤(1.60±0.75)次之,NF腺瘤評分最低(1.40±1.05),以上三種類型腫瘤,差異無統計學意義(p0.05)。上述結果被Western blot結果證實。4、垂體腺瘤組織中STAT3、p-STAT3和PTTG蛋白表達與患者性別、腫瘤大小、腫瘤侵襲性均無關(p0.05)。5、STAT3蛋白和PTTG蛋白表達正相關(rk=0.437, p0.01), p-STAT3蛋白和PTTG蛋白表達正相關(rk=0.322,p0.01)。6. STAT3 mRNA和PTTG mRNA在PRL、ACTH、GH和NF腺瘤均有表達,其中以GH腺瘤表達較高。結論1、STAT3 mRNA和蛋白在垂體腺瘤組織中均有表達。2、垂體腺瘤組織中存在STAT3的持續(xù)激活。3、STAT3、p-STAT3、PTTG在不同類型垂體腺瘤表達水平不一,在GH腺瘤表達水平最高,NF腺瘤表達水平最低。4、垂體腺瘤中活化的STAT3可能通過促進下游靶基因PTTG的轉錄來發(fā)揮作用。
[Abstract]:Pituitary adenomas are common benign intracranial tumors, accounting for about 10%-25% of intracranial tumors. They originate from highly differentiated anterior pituitary cells secreting one or more hormones. According to the pathological types, pituitary adenomas can be divided into growth hormone (GH) adenomas, prolactin (PRL) adenomas, adrenocorticotrophin (ACH). Adenomas (ACTH) adenomas, nonfunctioning (NF) adenomas, and other types of tumors have different clinical manifestations, such as acromegaly, amenorrhea/galactorrhea, Cushing's syndrome, or just compression of the surrounding structures. Regardless of the pathological type, most pituitary adenomas grow slowly, but a few pituitaries. Adenomas are characterized by malignant biological behaviors, such as aggressive rapid growth, no response to conventional surgery, radiation, and drug therapy. Therefore, judging the biological behavior of the tumor is helpful to judge the prognosis and treatment of the tumor. Therefore, further study of the molecular biological mechanism of pituitary adenoma development, discovery of tumor markers and their clinical significance will be of great significance for the prevention, diagnosis, prognosis evaluation and treatment of pituitary adenoma. Janus protein tyrosine kinase (JAK) / signal transduction and transcription activation Signal transducer and activator of transcription (STAT) signaling pathway is an extremely fast signal transduction pathway from extracellular to nuclear. At present, seven members of STAT family have been identified in mammals: STAT1, STAT2, STAT3, STAT4, STAT5a, STAT5b and STAT6. Among them, STAT3 is the most widely studied. STAT3 egg White is latent in the cytoplasm, and phosphorylated STAT3 becomes an activated form of STAT3 (p-STAT3), forming homologous or heterodimers, translocating to the nucleus and binding to specific promoters, inducing the expression of target genes (such as PTTG, VEGF, MMP2, Cyclin-Dl, etc.) to regulate cell proliferation, differentiation, apoptosis, angiogenesis and other processes, if STAT3 The expression and activation of STAT3 in pituitary adenoma tissues and the relationship between STAT3 and PTTG have not been reported at home and abroad.Objective To detect pituitary adenoma tissues by immunohistochemistry, Western blot and Real Time PCR. The expression levels of STAT3, p-STAT3. PTTG protein and mRNA in pituitary adenomas were analyzed. The relationship between STAT3, p-STAT3, PTTG and the clinicopathological factors of pituitary adenomas was analyzed. Whether the expression of STAT3 regulating PTTG was involved in the pathogenesis of pituitary adenomas was discussed. From May 2010 to June 2013, 80 patients with pituitary adenomas underwent surgical treatment in the Department of Neurosurgery, Peking Union Medical College Hospital. All surgical specimens were confirmed by clinical and pathological diagnosis. The tumor specimens included 20 PRL adenomas, 20 ACTH adenomas, 20 GH adenomas and 20 NF adenomas. Among them, 27 were males, 53 females, aged 17-58 years, with an average of 40 years. According to Knosp standard, grade III and IV were defined as invasive pituitary adenomas, grade I and II were defined as non-invasive adenomas, including 30 invasive adenomas and 50 non-invasive adenomas. Results 1. STAT3 protein was stained in the nucleus and cytoplasm of immunohistochemical staining. STAT3 protein was evaluated in GH adenomas. The score of STAT3 protein in PRL adenoma and ACTH adenoma was significantly higher than that in PRL adenoma (3.90 (+ 2.29)) ACTH adenoma (3.55 (+ 1.53)) and NF adenoma (1.85 (+ 1.73)). There was no significant difference in STAT3 protein between PRL adenoma and ACTH adenoma (p = 0.57). Results The Western blot results confirmed that. 2. P-STAT3 protein nuclear staining. P-STAT3 protein in GH adenoma had the highest immunohistochemical score (3.20 (+ 1.50). Compared with other three types of tumors, the difference was statistically significant (p0.01). PRL adenoma (1.50 (+ 1.23) and ACTH adenoma (1.15 (+ 1.39) were the second, and NF adenoma (0.85 (+ 0.7). There was no significant difference between the three types (p0.05). The above results were confirmed by Western blot. 3. The immunohistochemical PTTG protein cytoplasmic staining. PTTG protein in GH adenoma scored the highest (2.05.99), compared with other three types of tumors, the difference was statistically significant (p0.05). ACTH adenoma (1.60.05) and PRL adenoma (1.60.99). The expression of STAT3, p-STAT3 and PTTG protein in pituitary adenomas was not correlated with gender, tumor size and tumor invasion (p0.05). STAT3 protein and PTTG protein expression were positively correlated. (rk = 0.437, p0.01), p-STAT3 protein and PTG protein expression were positively correlated (rk = 0.322, p0.01). 6. STAT3 mRNA and PTG mRNA were expressed in PRL, ACTH, GH and NF adenomas, and GH adenomas were highly expressed. Conclusion 1. STAT3 mRNA and protein were expressed in pituitary adenomas. 2. There was a persistent activation of STAT3, STAT3, p-STAT3, and P-STAT3 in pituitary adenomas. The expression of PTTG was highest in GH adenomas and lowest in NF adenomas. Activated STAT3 in pituitary adenomas may play a role by promoting the transcription of PTTG.
【學位授予單位】：北京協和醫(yī)學院
【學位級別】：博士
【學位授予年份】：2015
【分類號】：R736.4

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本文編號：2220754