Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/92108
標題: Cdk5蛋白於膀胱癌及前列腺癌中扮演角色之探討
Investigation of the roles of Cdk5 protein in bladder cancer and prostate cancer
作者: Eugene Lin
林育慶
關鍵字: Cdk5;p35;膀胱癌;前列腺癌;Cdk5;p35;bladder cancer;prostate cancer
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摘要: 
Prostate cancer is the most common malignancy among men and bladder cancer is the fourth most common malignancy among men in the United States. In Taiwan, prostate cancer and bladder cancer also play an important role in male urinary tract malignancies. Recent studies have explored the roles of cyclin-dependent kinase 5 (Cdk5) and its activator p35 in cancer research. We previously suggested that Cdk5 regulates the proliferation of prostate and thyroid cancer cells; thus, the aim of this thesis is to investigate the relation between Cdk5/p35 protein expression and the biophysical function in prostate and bladder cancer cells.

Part I: The overexpression of cyclin-dependent kinase 5 and p35 in urinary bladder urothelial carcinoma and associated with bladder cancer cells migration activity

We collected bladder cancer specimens from 27 patients (admitted by Institutional Review Board of Chang Bing Show Chwen Memorial Hospital). The levels of Cdk5 and p35 protein were analyzed using immunohistochemical staining or western blotting and compared between tumors and adjacent non-cancerous bladder tissues from individual patient. Migration array was conducted using bladder cancer cell line T24 in while Cdk5 was overexpressed or inhibited by inhibitor. The data indicated that the levels of Cdk5 and p35 protein expression were higher in tumors compared with the adjacent normal bladder tissues in 18 out of 27 patients. In addition, Cdk5 overexpression increased the migration activity of bladder cancer cell line T24 and the migration can be inhibited by treating with the Cdk5 inhibitor, roscovitine (p < 0.05). We first report the overexpression of Cdk5 and p35 proteins in bladder cancer specimens. These findings suggest that Cdk5 might be a tumor marker of bladder cancer and respond to regulate cell migration. Thus, these findings should help future bladder cancer diagnosis and treatment.

Part II: Cyclin-dependent kinase 5 and p35 protein expressions were associated with prostate cancer grade and stage

Prostate cancer is the most frequently diagnosed cancer and second leading cause of cancer-related deaths in men. The roles of cyclin-dependent kinase 5 (Cdk5) and its activator p35 in cancer biology have been extensively studied. We previously demonstrated that Cdk5 may regulate proliferation of prostate and thyroid cancer cells. The aim of this study was to investigate the relationship between Cdk5 and p35 protein expressions and prostate cancer cell grade and stage. We collected 212 patient specimens of prostate cancer from commercial tissue array. Cdk5 and p35 protein levels were evaluated by immunohistochemical staining. The images were double-blind evaluated by two experts in accordance with a scoring system based on the intensity and distribution of staining signals. The correlations between Cdk5 and p35 protein levels and tumor grade or stages were analyzed using Fisher's exact test. The overexpression of Cdk5/p35 was significantly correlated with prostate cancer grade (p = 0.0277) as well as stage (p = 0.0150). A higher grade and stage of prostate cancer was associated with higher expressions of Cdk5 and p35 proteins. This study first identified overexpression of Cdk5 and p35 in prostate cancer using tissue array specimens, which implies that Cdk5 is a potential tumor marker of prostate cancer. These findings may help making early diagnosis of prostate cancer and in the development of new treatments in the future.

To sum up the conclusions of part I and part II, we indicate the role of Cdk5 and p35 in bladder cancer and prostate cancer. By investigating on the expression of Cdk5 and p35 proteins, we explored the potential roles of Cdk5 and p35 proteins in prostate cancer and bladder cancer. We look forward to the further cell biology evidence to demonstrate the mechanism of Cdk5 and p35 proteins in tumors of prostate and bladder.

前列腺癌為美國男性排名第一常被發現之癌症,而膀胱癌為美國男性排名第四常被發現之癌症;在台灣,膀胱癌與前列腺癌亦為男性泌尿系統重要之癌症。於近期癌症生物學的研究中,Cdk5蛋白及其活化子p35已被廣泛的探討,根據以往本團隊發表之文獻顯示,Cdk5蛋白可影響前列腺癌和甲狀腺癌細胞的增殖;因此,本研究希望探討Cdk5和p35蛋白表現與前列腺癌及膀胱癌之間的關係,希望這些研究結果應該有助於促進未來的前列腺癌及膀胱癌的診斷和治療。

第一部份: Cdk5蛋白質在膀胱癌之表現及與膀胱癌細胞遷移活性之關係

首先,我們通過彰濱秀傳紀念醫院人體試驗委員會審核,並收集了27個新鮮臨床膀胱癌標本,進一步利用西方墨點法分析Cdk5和p35的蛋白表現,並將腫瘤組織和腫瘤旁非癌化膀胱組織之間進行比較其腫瘤細胞惡性度和Cdk5及p35的蛋白表現的相關性;另一方面,我們也採用膀胱癌細胞株T24、RT4和TSGH8301,將其Cdk5基因過度表現或抑制來進行細胞侵犯性實驗。結果顯示,27位病患檢體中有18位的膀胱癌標本之Cdk5和p35的蛋白表現高於配對的腫瘤旁正常膀胱組織;另外Cdk5蛋白的過度表現可增加膀胱癌T24細胞的遷移活性;而這個現象可以透過Cdk5的抑制劑roscovitine加以抑制,此為膀胱癌中Cdk5和p35蛋白的過度表現與細胞活動性之間的關係首次被發現。由上述結果可知,Cdk5未來可能可以成為膀胱癌的腫瘤標記物,且抑制Cdk5蛋白之活性可以抑制膀胱癌細胞遷移反應。希望這些研究結果應該有助於促進未來的膀胱癌的診斷和治療。

第二部份: Cdk5及p35蛋白之過度表現與前列腺癌細胞惡性度及期別相關

前列腺癌是美國男性最常見的癌症和第二大癌症相關死亡的原因。Cdk5蛋白及活化子p35的作用於癌症生物學已被廣泛研究;根據我們之前發表之文獻指出,Cdk5基因可調節前列腺癌和甲狀腺癌細胞的增生。本研究的目的是觀察Cdk5和p35蛋白表現與前列腺癌的細胞惡性度和分期之間的關係。我們利用市售的組織微陣列切片觀察了212個前列腺癌組織標本。Cdk5和p35蛋白表現以組織免疫染色評估。這些影像由兩位專家以雙盲之模式,依據染色信號強度和分佈的評分系統加以評估。Cdk5和p35蛋白表現與腫瘤的細胞惡性度和分期之間的相關性採用Fisher精確檢驗統計。結果顯示,Cdk5和P35蛋白的表現與前列腺癌細胞惡性度(p = 0.0277)或分期(p = 0.0150)呈現顯著地相關性,亦即較高細胞惡性度和分期的前列腺癌有較高的Cdk5和P35蛋白之表現。由此推論, Cdk5可能是前列腺癌的一個潛在腫瘤標記物,配合本團隊已發表的文獻可知,抑制Cdk5蛋白表現或活性可抑制前列腺癌的生長。希望這些發現未來有助於使前列腺癌的早期診斷和發展新的治療方法。
綜合第一及第二部份的研究結論,我們確立了Cdk5及p35蛋白在膀胱癌及前列腺癌之重要性。在腫瘤研究上,我們以膀胱癌及前列腺癌細胞來探討Cdk5及p35蛋白在泌尿系癌症之表現,期待後續進一步之分子細胞研究能解答其機轉,並確立其在泌尿系癌症之標的角色。
URI: http://hdl.handle.net/11455/92108
Rights: 同意授權瀏覽/列印電子全文服務,2017-06-30起公開。
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