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標題: | 研究LCN2蛋白質與子宮內膜疾病之相關性 Characterization of the relationship between LCN2 and endometrial diseases |
作者: | Chi-Jr Liao 廖啟志 |
指導教授: | 朱善德 |
關鍵字: | 子宮內膜異位症,子宮內膜增生症,子宮肌腺症,LCN2蛋白質, endometriosis,endometrial hyperplasia,adenomyosis,LCN2, |
出版年 : | 2014 |
學位: | 博士 |
摘要: | 子宮肌腺症、子宮內膜增生症以及子宮內膜異位症均是子宮內膜組織的病變,屬於發生率較高的婦科疾病。目前認為子宮內膜增生症屬於高危險疾病,是子宮內膜癌的前兆,相較子宮肌腺症則歸類於低危險的組織增生現象。子宮內膜異位症則除了與卵巢癌有高度相關外,與其他癌症似乎也有關聯性,而這些症狀的發生可能與組織中的基因調節有關。LCN2蛋白質為疏水性小分子結合蛋白家族成員,屬於急性反應蛋白,在生理週期中子宮內膜組織會受到雌激素的作用而分泌LCN2蛋白質,當細胞處於壓力狀態下,如環境養分貧瘠也會誘導細胞產生LCN2蛋白質,並且在不同的濃度即時間點具有誘導或是抑制細胞凋亡的不同現象。本研究分成兩部分,首先了解子宮肌腺症與子宮內膜增生症兩者組織內LCN2蛋白質的表現量與上皮細胞轉型間質細胞因子彼此間的關聯。另一部分則是根據此一相關性研究之結果,分析LCN2蛋白質在子宮內膜異位症發生的初期可能扮演角色探討。
分析人類子宮肌腺症與子宮內膜增生症檢體內LCN2與癌症相關的基因表現量,在蛋白質或基因表現的層級上,子宮內膜增生症LCN2的表現量均顯著高於子宮肌腺症。進一步利用免疫組織染色法確認LCN2蛋白質高度表現在子宮內膜增生症的腺上皮細胞而非基質細胞。LCN2基因的表現量與COX2、CDH1呈現正相關,與vimentin、CTNNB1呈現負相關,初步推論在子宮內膜增生症所高度表現的LCN2可能具有避免子宮內膜增生組織轉變成為惡性組織之機制。 進一步利用動物模式了解LCN2蛋白質在子宮組織病變中可能扮演的角色。子宮內膜異位症之動物模式是較易建立的,於是利用移植子宮組織的方式建立子宮內膜異位小鼠模式,分析Lcn2蛋白質於血液、腹腔液與異位組織中不同時間點的表現量,並觀察Lcn2蛋白質在初級內膜上皮細胞培養實驗中對細胞生長與型態的影響。由小鼠動物模式證實Lcn2蛋白質與子宮內膜異位組織的發育成正相關性,以抗Lcn2蛋白質抗體中和腹腔內Lcn2蛋白質之作用,則可以觀察到子宮內膜異位組織體積變小,體外的初級子宮內膜上皮細胞在營養缺乏的環境下會提高Lcn2蛋白質的表現量,並且使細胞型態趨向間質細胞型,同時也會增加細胞的移動與侵蝕能力。研究結果顯示Lcn2蛋白質與子宮內膜增生症具高度關聯性,並可能參與子宮內膜異位症的初期發展,因此LCN2有做為此疾病生物性指標之潛力,未來並希望可能成為臨床上治療的標的。 Adenomyosis, endometrial hyperplasia and endometriosis are the most common gynecological disorders that result from the abnormal growth of the endometrium. Adenomyosis is associated with benign lesions, but endometrial hyperplasia is a high-risk disease and is considered to be a precursor of endometrial carcinoma. Endometriosis is not only related to ovarian cancer but is also highly associated with other cancers. The pathogenesis of these disorders is associated with gene regulation. LCN2 is a member of the lipocalin family and is also a kind of acute-phase protein. Estrogen can induce the secretion of LCN2 protein in the endometrium during the menstrual cycle. Stress, such as cellular starvation, may also cause the production of LCN2 protein. LCN2 has the ability to elicit or inhibit the process of apoptosis under different conditions. Two major topics were investigated in this study. The first was the relationship between the protein expression level of LCN2 and the epithelial-mesenchymal transition (EMT) in adenomyosis and endometrial hyperplasia. Next, according to the relativity, functional characterization of LCN2 during the initial endometriosis was verified. Analysis of the mRNA and protein expression level of LCN2 and other cancer-related genes in adenomyosis and endometrial hyperplasia showed that LCN2 expression was significantly higher in endometrial hyperplasia. Furthermore, via immunohistochemistry, the overexpression of LCN2 protein in the glandular epithelial cells but not the stroma cells in endometrial hyperplasia was confirmed. The expression pattern of LCN2 correlated positively with cyclooxygenase 2 (COX2) and CDH1 and negatively with vimentin and CTNNB1. This suggests that the increased LCN2 expression in patients with endometrial hyperplasia may participate in the prevention of the transition from hyperplasia to carcinoma. The animal model was used to verify the possible role of LCN2 in these endometrial disorders. Compared to adenomyosis and endometrial hyperplasia, the animal model of endometriosis is easier to establish. Autotransplantation of mouse endometrial tissue into the peritoneum was used as an animal model to analyze the Lcn2 protein expression level in the blood, peritoneal fluid and ectopic tissues during the pathogenesis of endometriosis and to study the cellular behavior in primary mouse endometrial epithelium cells affected by Lcn2 protein. The role of LCN2 in the pathology of endometriosis was investigated. The growth of ectopic endometrial tissue was inhibited by the Lcn2 antibodies in mice with implanted endometrial tissue. The increase of EMT, cellular migration and invasion ability correlated with the up-regulation of LCN2 expression in primary endometrial epithelial cells under the influence of nutrient deprivation. Our results reveal that LCN2 is highly related to endometriosis and might be involved in the formation of endometriosis tissues. This suggests that LCN2 could be a potential biomarker and target for clinical therapy in treating hyperplasia and endometriosis. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58431 |
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