垂体泌乳素腺瘤中雌激素受体信号传导通路影响细胞增殖和泌乳素分泌作用机制及其药物治疗评价体系的研究

Prolactinoma is regulated both by dopaminergic pathway and estrogen (E) /estrogen receptor (ER) signaling pathway. Many disadvantages exist in the treatment of prolactinoma. Great importance should be attached to anti-estrogen or ER antagonists treatment. The mechanism of E/ER signaling pathway in prolactinomas is still unclear, and the endocrine therapy system of prolactinoma based on anti-estrogen/ER has not been established yet. The current literature and our previous study have found that the expression of CaBP-9k is closely related to ER, and CaBP-9k/ER may play an important role in prolactinoma tumorigenesis and development, however, the mechanisms remain unclear. In this study, we consider CaBP-9k as the breakthough point, using the signal transduction inhibitors, luciferase reporter gene and immunoprecipitation techniques to analyze the regulation mechanisms of estrogen genomic and non-genomic pathways in prolactinoma cell proliferation and prolactin (PRL) secretion, clarify the relationship between CaBP-9k and ER, illustrate the possibilty of using CaBP-9k and ER as selecting molecular markers for anti-estrogen/ER antagonists treatment, and illuminate that E/ER and DA/D2R pathways are competitor in Pit-1 gene transcription process, which may be the pivotal target in prolactinoma tumorigenesis and development. Futher, we will use the CaBP-9k gene silencing and over-expressing prolactinoma cells, and gene knockout mice for in vitro and in vivo experiments to confirm that CaBP-9k plays a pivotal role in E/ER signaling pathways. Eventually, the study will reveal the prolactinoma tumorigenesis mechanisms and the fuctional mechanism of E/ER signaling pathways in prolactinoma, establism an evaluation system for anti-estrogen/ER antagonists endocrine treatment, and provide a novel therapeutic aprroach for pituitary prolactinoma treatment.

泌乳素腺瘤受多巴胺能途径和雌激素(E)/雌激素受体(ER)途径双重调节。现有治疗手段存在诸多弊端，抗E/ER治疗应当受到重视。但E/ER途径还不明确；缺乏抗E/ER药物治疗的筛选体系。文献和前期工作表明，在E/ER途径，CaBP-9k表达与ER密切相关，二者可能促进肿瘤发展，但机制不清楚。本课题拟采用信号传导干扰、荧光素酶报告基因、免疫沉淀等方法，以CaBP-9k为切入点，分析E两种途径对细胞增殖和PRL分泌的调控机制；ER与CaBP-9k相关性；阐明抗E/ER治疗、ER与CaBP-9k作为筛选抗E/ER治疗指标的可行性；E/ER、DA/D2R途径对Pit-1位点的竞争性抑制关系。采用CaBP-9k基因沉默和过表达细胞系及基因敲除鼠，揭示CaBP-9k在E/ER通路中的作用。通过本研究揭示泌乳素腺瘤发病机制、E/ER在泌乳素腺瘤中的作用机理，建立抗E/ER治疗的评价体系，开辟新的治疗途径。

泌乳素腺瘤受多巴胺能途径和雌激素(E)/雌激素受体(ER)途径双重调节。现有治疗手段存在诸多弊端，抗E/ER治疗应当受到重视。但E/ER途径还不明确；缺乏抗E/ER药物治疗的筛选体系。文献和前期工作表明，在E/ER途径，CaBP-9k表达与ER密切相关，二者可能促进肿瘤发展，但机制不清楚。本课题采用信号传导干扰、荧光素酶报告基因、免疫沉淀等方法，以CaBP-9k为切入点，揭示CaBP-9k在E/ER通路中的作用，分析E两种途径对细胞增殖和PRL分泌的调控机制；ER与CaBP-9k相关性；阐明CaBP-9k在泌乳素腺瘤细胞增殖与PRL分泌中的重要作用，垂体泌乳素腺瘤中E信号转导通路基因组途径(nER-ERE途径)和非基因组途径(mER或G蛋白偶联受体与下游PI3K/AKT/NK-κB、ERK/MAPK途径)对CaBP-9k表达的调控机制， ER与CaBP-9k对泌乳素腺瘤细胞增殖、凋亡与PRL分泌的作用机制，以及ER与CaBP-9k表达的相关性及二者与雌激素受体拮抗剂抑制作用的关系，并通过对人垂体泌乳素腺瘤组织标本ER与CaBP-9k表达与临床参数分析最终揭示ER与CaBP-9k联合检测可以作为筛选垂体泌乳素腺瘤内分泌治疗类型的分子标志，与此同时为泌乳素腺瘤以及其他类型雌激素相关性肿瘤发病机制的研究和新型药物开发寻找新的突破。泌乳素腺瘤发病机制中的两个重要信号传导途径E/ER与DA/D2R在Pit-1这个作用点中存在竞争性抑制关系可能为泌乳素腺瘤发病的关键，填补了垂体泌乳素腺瘤发病机制研究领域的空缺。