垂体泌乳素腺瘤诊断和耐药机制相关研究及他汀类药物治疗初探

发布时间：2018-05-01 09:09

本文选题：泌乳素腺瘤 + PRL　；参考：《北京协和医学院》2015年博士论文

【摘要】：研究背景：垂体泌乳素腺瘤是最常见的功能性垂体腺瘤,约占成人腺瘤的40%-50%。其典型的临床表现为高泌乳素血症、鞍区周围组织压迫症状、垂体前叶部分功能受损等。首选多巴胺受体激动剂治疗,可对90%的患者缓解症状。目前中外垂体泌乳素腺瘤相关指南认为血清泌乳素水平高于200ng/mL的垂体腺瘤明确诊断泌乳素腺瘤,而高于100ng/mL则主要考虑泌乳素腺瘤的诊断；但是对于泌乳素水平高于正常但低于100ng/mL的患者诊断尚不明确。而10%对多巴胺受体激动剂不敏感的患者称之为耐药性泌乳素腺瘤,目前对于泌乳素腺瘤耐药机制的研究及治疗是前沿领域,其中血管新生是否是可能的耐药机制及潜在治疗靶点尚待研究；而他汀类药物作为新兴的抗肿瘤药物在垂体泌乳素腺瘤中是否存在作用仍有待研究。本文主要正对泌乳素腺瘤诊断和治疗中的相关问题予以探索和分析。研究目的：1)明确垂体泌乳素腺瘤诊断的泌乳素水平临界值的范围。2)观察耐药性垂体泌乳素腺瘤与敏感性垂体泌乳素腺瘤在血管新生方面是否存在差异。3)初步探究他汀类药物对于垂体泌乳素腺瘤MMQ和GH3细胞系的作用及潜在的作用机制。研究方法：1)采用IPP软件半定量分析不同泌乳素水平的垂体腺瘤免疫组化PRL染色阳性程度的差异,并为确立诊断泌乳素腺瘤的血清泌乳素水平临界值提供依据。2)采用免疫组化观察耐药性垂体泌乳素腺瘤及敏感性垂体泌乳素腺瘤标本血管新生相关因子表达量的差异,如血管内皮生长因子(VEGF)和CD31；以及胞外基质蛋白如MMP2和MMP9表达量的差异。3)使用不同浓度的辛伐他汀处理大鼠MMQ和GH3细胞系的作用,采用CCK8、流式细胞学、Western Blotting和Elisa等方法分析其对细胞增殖、凋亡、PRL表达等的作用及潜在的分子机制。结果：1)泌乳素水平高于100ng/mL的垂体腺瘤,与泌乳素水平高于正常但小于100ng/mL的垂体腺瘤相比,免疫组化PRL表达量明显高于后者；2)耐药性垂体泌乳素腺瘤的VEGF表达量高于敏感性垂体泌乳素腺瘤组,但二者在MMP2、MMP9和CD31方面的表达量基本没有差异。3)耐药性垂体泌乳素腺瘤VEGF表达量的高低与Ki67数值相关,与肿瘤大小、泌乳素水平无关。4)他汀类药物可在MMQ和GH3细胞系中抑制增殖、诱导凋亡、抑制PRL的分泌并抑制VEGF蛋白表达量。结论：1)血清泌乳素水平高于100ng/mL的垂体腺瘤首先考虑诊断垂体泌乳素腺瘤。2)耐药性垂体泌乳素腺瘤VEGF的表达量与细胞增殖程度相关,可能与难治性泌乳素腺瘤或侵袭性泌乳素腺瘤相关。3)他汀类药物可能作为耐药性垂体泌乳素腺瘤的潜在治疗药物,更多分子机制有待研究。
[Abstract]:Background: pituitary prolactin adenomas are the most common functional pituitary adenomas, accounting for 40-50 percent of adult adenomas. The typical clinical manifestations were hyperprolactinemia, compression of periSellar tissues and impaired anterior pituitary function. The first choice of dopamine receptor agonists is to relieve symptoms in 90% of patients. At present, the related guidelines of pituitary prolactin adenoma in China and abroad think that the pituitary adenoma with higher serum prolactin level than 200ng/mL has definite diagnosis of prolactin adenoma, but higher than 100ng/mL mainly considers the diagnosis of prolactin adenoma. But the diagnosis of prolactin levels higher than normal but lower than 100ng/mL remains unclear. 10% of the patients who are insensitive to dopamine receptor agonists are called drug-resistant prolactin adenomas. Whether angiogenesis is a possible drug resistance mechanism and a potential therapeutic target remains to be studied, and whether statins are a new antitumor agent in pituitary prolactin adenoma remains to be studied. This article is mainly about the diagnosis and treatment of prolactinoma. Objective: to determine the range of critical value of prolactin level in diagnosis of pituitary prolactin adenoma. (2) to observe whether there are differences in angiogenesis between drug-resistant pituitary prolactin adenoma and sensitive pituitary prolactin adenoma. Effect of statins on MMQ and GH3 cell lines of prolactinoma and its potential mechanism. Methods IPP software was used to semi-quantitatively analyze the difference of immunohistochemical PRL staining in pituitary adenomas with different prolactin levels. In order to establish the critical value of serum prolactin level in the diagnosis of prolactinoma, the expression of angiogenic factors in drug-resistant pituitary prolactin adenoma and sensitive pituitary prolactin adenoma was observed by immunohistochemistry. For example, vascular endothelial growth factor (VEGF) and CD31, and the difference in expression of extracellular matrix proteins such as MMP2 and MMP9.) the effects of simvastatin on rat MMQ and GH3 cell lines treated with different concentrations of simvastatin were observed. The effects of CCK8, flow cytometry, Western Blotting and Elisa on cell proliferation, apoptosis and expression of PRL were analyzed and their potential molecular mechanisms were analyzed. Results the prolactin level in pituitary adenomas with 100ng/mL was higher than that in pituitary adenomas with higher prolactin level than in pituitary adenomas with higher prolactin level but smaller than 100ng/mL. The expression of VEGF in prolactin adenoma was significantly higher than that in the sensitive pituitary prolactinoma group, and the expression of VEGF in the resistant pituitary prolactinoma was significantly higher than that in the sensitive pituitary prolactinoma group. However, there was no difference in the expression of MMP2, MMP9 and CD31 between them. 3) the level of VEGF expression in drug-resistant pituitary prolactin adenoma was correlated with the value of Ki67, but not with tumor size, prolactin level. 4) statins could inhibit proliferation in MMQ and GH3 cell lines. Apoptosis was induced, PRL secretion was inhibited and VEGF protein expression was inhibited. Conclusion in pituitary adenomas whose serum prolactin level is higher than that of 100ng/mL, the first consideration is to diagnose prolactin adenoma. 2) the expression of VEGF in drug-resistant pituitary prolactin adenoma is correlated with cell proliferation. May be associated with refractory prolactin adenoma or invasive prolactin adenoma. 3) statins may be a potential therapeutic drug for drug-resistant pituitary prolactin adenoma, more molecular mechanisms need to be studied.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R736.4

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