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研究生：	張宜雯 Chang, Yi-Wen
論文名稱：	CDCP1藉由the c-Src/PKC-delta路徑促進尿路上皮癌細胞的幹細胞特性 CUB Domain-containing Protein 1 promotes stemness of urothelial carcinoma cells via the c-Src/PKC-delta pathway
指導教授：	黃暉升 Huang, Huei-Sheng
學位類別：	碩士 Master
系所名稱：	醫學院 - 醫學檢驗生物技術學系 Department of Medical Laboratory Science and Biotechnology
論文出版年：	2023
畢業學年度：	111
語文別：	英文
論文頁數：	53
中文關鍵詞：	尿路上皮癌、腫瘤幹細胞、抗藥性
外文關鍵詞：	CUB Domain-containing Protein 1, Urothelial carcinoma, Cancer stem cells, Drug resistance
相關次數：	點閱：59 下載：0
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尿路上皮癌 (urothelial carcinoma, UC) 是指在泌尿系統上皮發生的癌症，臨床上約有90%的UC病人皆屬於膀胱癌。根據美國統計男性十大常見的癌症中，尿路上皮癌位居第四名。在臨床上膀胱癌多以化學療法等方式來做治療，然而卻有越來越多的病人再給予化療藥物後產生抗藥性。因此了解膀胱癌病人的抗藥性機轉相當重要，並能提供臨床病患一個更好的治療策略。CUB Domain-containing Protein 1 (CDCP1)是一個細胞表面的跨膜蛋白，在許多癌症中發現CDCP1高度表現，並影響癌症的進展，而CDCP1在血液細胞中也被認為是一個重要的幹細胞標誌。根據一些報導指出，癌症幹細胞(Cancer stem cells)具有自我複製及分化的能力，與治療藥物抗性息息相關。但確切的機制仍不清楚，研究相關機制有助於提升膀胱癌病人的化療成效。本篇研究主軸為探討CDCP1在具有幹細胞特性的尿路上皮癌細胞中的角色。在我們的實驗結果中證明了T24過度表達CDCP1細胞與T24相比有較高的幹細胞特性，且表現了幹細胞相關標誌KLF4及SOX2、也影響抗藥性的產生。為了進一步探討原因，我們針對T24-flCDCP1使用特異性的Src及PKCδ抑制劑後發現幹細胞樣的表型及細胞遷移能力明顯下降。除此之外，我們將Gemcitabine及PKCδ抑制劑(Rottlerin)合併使用後減少了T24-flCDCP1的細胞存活率。總結來說，CDCP1/c-Src/PKCδ影響了幹細胞生長特性並且是影響抗藥性產生的原因，抑制這條細胞傳遞訊路徑可能有助於提升膀胱癌細胞中的化療抗性。

Urothelial carcinoma (UC) is the fourth most common cancer in men in the United States of America. Chemotherapy plays a vital role in the treatment of UC. More and more UC patients express no response to chemotherapy, high rate of disease recurrence and exhibition of drug resistance are significant challenges for the treatment. It is critical to improve the efficacy of chemotherapy in UC patients. Accumulating evidence indicates that cancer stem cells (CSCs) are correlated with self-renewal recurrence and drug resistance of tumors, including UC. However, the exact mechanism remains unclear. CUB Domain-containing Protein 1 (CDCP1) is a cell surface transmembrane protein overexpressed in various human cancers to drive cancer progression. CDCP1 has also been identified as a stem cell marker of hematopoietic cells. The study aims to elucidate the role of CDCP1 in the stemness of UC and resultant chemoresistance. Our results demonstrated that CDCP1-overexpressed T24 cells (T24-flCDCP1) revealed more stem cell-like phenotype than T24 cells, including sphere formation, expression of stem cell markers KLF4 and Sox2, and chemoresistance. To further elucidate the mechanisms, we treated T24-flCDCP1 with a specific Src inhibitor and PKCδ inhibitor (Rottlerin), and the inhibition of stem cell-like phenotype and cellular migration was observed. In addition, combined treatment of Gemcitabine with Rottlerin enhanced the cytotoxicity of T24-flCDCP1. We conclude that CDCP1/c-Src/PKCδ contributes to the stemness and chemoresistance of UC cells. Inhibition of the pathway might improve the chemoresistance of UC cells.

Abstract I
摘要 II
誌謝 III
Contents Ⅳ
Introduction 1
Hypothesis and Specific Aim 6
Materials and Methods 7
Results 20
CDCP1 increases stemness in UC cells 20
CDCP1 promotes cell migration and invasion in UC cells 21
Stemness of T24-flCDCP1 can be inhibited by Src and PKCδ inhibitors 22
Cell migration and invasion ability in T24-flCDCP1 can be inhibited by Src 
and PKCδ inhibitors 23
CDCP1 contributes to the chemoresistance of UC cells 24
A combination of Rottlerin/Gemcitabine enhanced the cytotoxicity of 
T24-flCDCP1 25
Discussion 26
Figures 31
Figure 1. CDCP1 increases stemness in UC cells 31
Figure 2. CDCP1 promotes cell migration and invasion in UC cells 34
Figure 3. Stemness of T24-flCDCP1 can be inhibited by Src and PKCδ inhibitors 36
Figure 4. Cell migration and invasion ability in T24-flCDCP1 can be inhibited by Src 
and PKCδ inhibitors 39
Figure 5. CDCP1 contributes to the chemoresistance of UC cells 41
Figure 6. A combination of Rottlerin/Gemcitabine enhanced the cytotoxicity of 
T24-flCDCP1 44
References 46
Appendix 50
Appendix 1. The structure of CDCP1 50
Appendix 2. Potential applications of CDCP1 in cancer 51
Appendix 3. Targeting proteolytic neoepitopes could provide therapeutic strategies 52
Appendix 4. Main characteristics of cancer stem cells(CSCs) 53
                                    

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校內：2024-01-01公開
校外：2024-01-01公開

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