雄性荷爾蒙接受體是上泌尿道泌尿上皮癌的潛在治療標的

中國醫藥大學機構典藏 China Medical University Repository, Taiwan > 醫學院 > 臨床醫學研究所 > 博碩士論文 > Item 310903500/50306

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題名:	雄性荷爾蒙接受體是上泌尿道泌尿上皮癌的潛在治療標的 Androgen Receptor Is a Potential Therapeutic Target for Upper Urinary Tract Urothelial Cell Carcinoma
作者:	謝登富;Teng-Fu Hsieh
貢獻者:	臨床醫學研究所博士班
關鍵詞:	雄性荷爾蒙;雄性荷爾蒙接受體;上泌尿道;泌尿上皮癌;治療標的;Androgen;Androgen receptor;Upper urinary tract;Urothelial cell carcinoma;Therapeutic target
日期:	2013-07-03
上傳時間:	2013-10-02 11:22:01 (UTC+8)
出版者:	中國醫藥大學
摘要:	在台灣，上尿路尿路上皮癌是最常見的上尿路癌症。這個疾病的治療方式有根除性腎臟及輸尿管切除手術或化療，但結果都不理想。同時這些治療對患者的生活都會產生很大的影響。許多研究發現，性別差異影響疾病發生率是一個重要特點。於是，我們假設，雄激素受體 (AR) 在這個上尿路尿路上皮癌扮演一個重要的角色。為了印證這個假設，我們設計了以下四個實驗目標。目標1：檢測上尿路尿路上皮癌組織性激素受體的表現及測試上尿路尿路上皮癌細胞性激素受體的功能。目標2：利用人類上尿路尿路上皮細胞癌培養細胞來當作預後預測指標和藥物敏感試驗。目標3：檢測AR在上尿路尿路上皮腫瘤細胞能否增加化療藥物的細胞毒作用。目標4：檢測AR對於上尿路尿路上皮癌細胞的遷移和侵襲的作用。在目標1，我們評估性激素受體於上尿路尿路上皮癌表現。採用免疫組織化學檢測性激素受體在上尿路尿路上皮癌的表現數量。 XTT試驗用於評估細胞對抗癌藥物治療效果。傷口癒合檢測方式以確定細胞的遷移能力。在輸尿管的上尿路尿路上皮癌標本中發現於表淺或低級別腫瘤有較高的AR表達。相比之下，ERβ表達差異不大。此外，AR會降低上尿路上皮癌對於化療藥物的敏感性，但會增加細胞的遷移能力。在目標2，我們取得手術後的上尿路尿路上皮癌腫瘤組織細胞，建立了個別原代培養的細胞系，我們使用E-cadherin蛋白和Ki-67來證實這是上尿路尿路上皮癌腫瘤組織細胞，進而進行化療藥物敏感性測試。觀察藥物在這些細胞中的毒殺作用。結果顯示這個方式可能可以來預測預後，並為化療的療效評估提供了一個新的平台。在目標3，我們發現AR在上尿路尿路上皮癌細胞（BFTC909）的過度表達可以減少化療藥物的細胞毒作用並協助上尿路尿路上皮癌細胞抵抗化療藥物。同時會增加化療藥物抗藥性的基因如ABCG2和ATP-binding cassette half-transporter的表現。此外，使用的AR的拮抗劑 ASC-J9 ，則可以減少AR增加的影響。在目標4，我們使用BFTC 909細胞來研究。 AR的過度表達會促進 BFTC909 細胞的遷移和侵襲。我們使用PCR定量和Western blot分析來研究遷移/侵襲相關的基因表達。結果證明，AR增強上尿路尿路上皮癌細胞遷移和侵襲的基因如MMP-9和COX-2的表現。隨後，阻斷MMP-9和COX-2信號就可以抑制AR增強的細胞遷移和侵襲。我們總結整個研究，可以發現AR影響上尿路尿路上皮癌細胞的增殖，遷移和侵襲。它還可以增加化療藥物的細胞毒作用。因此，AR可能可以作為上尿路尿路上皮癌的一種新的生物標誌物和潛在的治療標靶。 In Taiwan, upper urinary tract urothelial cell carcinoma is the most common cancer in upper urinary tract. Remove the disease kidney with ureter or chemotherapy are the treatment of the upper urinary tract urothelial cell carcinoma, but the result are dispointment. The upper urinary tract urothelial cell carcinoma has great impact to the patients. Many studies indicate that gender difference is an important characteristic in upper urinary tract urothelial carcinomas. We offer a hypothesis that the androgen receptor plays a role in this cancer. To this purpose, we design the following four aims. Aim 1: To assay the expression and actions of androgen receptor in upper urinary tract urothelial carcinoma tissues and the primary cultured cells. Aim 2: To assay the potential use of primary human upper urinary tract urothelial cell carcinoma cultured cells for prognostic indicators and chemosensitivity test. Aim 3: To assay the androgen receptor role in the cytotoxic effects of chemotherapeutic drugs in upper urinary tract urothelial carcinoma cells. Aim 4: To assay the androgen receptor role in the migration and invasion of upper urinary tract urothelial carcinoma cells. In aim 1, we aimed to evaluate sex hormone receptors expression in upper urinary tract urothelial carcinomas of ureter and renal pelvis with different tumor stages and grades as well as their possible roles in tumor progression. Immunohistochemistry was used to assay the expression of AR and ERs in the primary upper urinary tract urothelial carcinomas. XTT viability test In Taiwan, upper urinary tract urothelial cell carcinoma is the most common cancer in upper urinary tract. Remove the disease kidney with ureter or chemotherapy are the treatment of the upper urinary tract urothelial cell carcinoma, but the result are dispointment. The upper urinary tract urothelial cell carcinoma has great impact to the patients. Many studies indicate that gender difference is an important characteristic in upper urinary tract urothelial carcinomas. We offer a hypothesis that the androgen receptor plays a role in this cancer. To this purpose, we design the following four aims. Aim 1: To assay the expression and actions of androgen receptor in upper urinary tract urothelial carcinoma tissues and the primary cultured cells. Aim 2: To assay the potential use of primary human upper urinary tract urothelial cell carcinoma cultured cells for prognostic indicators and chemosensitivity test. Aim 3: To assay the androgen receptor role in the cytotoxic effects of chemotherapeutic drugs in upper urinary tract urothelial carcinoma cells. Aim 4: To assay the androgen receptor role in the migration and invasion of upper urinary tract urothelial carcinoma cells. In aim 1, we aimed to evaluate sex hormone receptors expression in upper urinary tract urothelial carcinomas of ureter and renal pelvis with different tumor stages and grades as well as their possible roles in tumor progression. Immunohistochemistry was used to assay the expression of AR and ERs in the primary upper urinary tract urothelial carcinomas. XTT viability test was applied to evaluate cell responses for anticancer drug treatment. Wound healing assay was performed to determine cell migration abilities. AR and ERβ immunoreactivities were observed in both upper urinary tract urothelial carcinomas, but ERα was not detected in either upper urinary tract urothelial carcinomas. In upper urinary tract urothelial carcinomas of ureter specimens, higher AR expression was found in superficial or lower grade tumors. In contrast, little difference of ERβ expression was found in superficial versus muscle-invasive tumor stages or low grades versus high grades in upper urinary tract urothelial carcinomas of ureter specimens. Furthermore in the primary cultured cells from upper urinary tract urothelial carcinomas specimens, the addition of functional AR reduced cell chemosensitivity, but increased cell migration. These results provide the first data showing the expression patterns of sex hormone receptors in both renal pelvis and ureter upper urinary tract urothelial carcinomas. From results, we concluded that there is a positive correlation for higher AR expression found in superficial or low-grade upper urinary tract urothelial carcinomas s of ureter and identified the functional roles of AR in upper urinary tract urothelial carcinomas progression. In aim 2, we have obtained cells from upper urinary tract urothelial carcinomas tumor tissues after surgery and established individual primary cultured cell lines, which were used to evaluate E-cadherin and Ki-67 proliferation marker expression and their chemosensitivity to chemotherapeutic drugs. Differential Ki-67 expression and chemosensitivity were observed in these primary cultured cell lines, suggesting these types of upper urinary tract urothelial carcinomas cell lines could provide a platform for determining prognostic makers and evaluating the efficacy of chemotherapy. In conclusion, primary cultured cell lines from individual patients will be a great tool for evaluating and determining each individual’s personalized chemotherapy course and for testing and screening new chemotherapeutic agents against upper urinary tract urothelial carcinomas. In aim 3, AR overexpression in upper urinary tract urothelial carcinomas cells (BFTC 909) was identified to reduce the cytotoxic effect of chemotherapeutic drugs, including doxorubicin, cisplatin and mitomycin C and protected cells from drug-induced death. The expression of ABCG2, an ATP-binding cassette half-transporter associated with multidrug resistance, was increased in AR-overexpressing BFTC cells. In addition, use of the AR degradation enhancer, ASC-J9?, repressed the AR effect on increasing cell viability. In aim 4, we investigated the role of the AR in upper urinary tract urothelial carcinomas by using upper urinary tract urothelial carcinomas -derived BFTC 909 cells. The overexpression of AR promotes the migration and invasion of BFTC 909 cells. Expression of migration/invasion-related genes was increased in BFTC 909 cells overexpressing AR determined by qPCR and western blot analyses. The results showed that AR-enhanced migration and invasion of upper urinary tract urothelial carcinomas cells are linked to the upregulation of the matrix-degrading enzyme MMP-9 and cyclooxygenase (COX)-2. Subsequently, the blocking of MMP-9 and COX-2 signaling by inhibitors suppressed AR-enhanced cell migration and invasion. The results of the present study provide evidence for the first In the conclusions, the androgen receptor plays a role in the proliferation, migration and invasion of upper urinary tract urothelial carcinoma cells. It also can increase cytotoxic effects of chemotherapeutic drugs. Thus, the androgen receptor may also serve as a novel biomarker and potential therapeutic target for upper urinary tract urothelial carcinomas.
顯示於類別:	[臨床醫學研究所] 博碩士論文

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