- 學位論文
趨化激素CXCL14（BRAK）與人類腎細胞癌轉移的分子機制
The molecular mechanism of chemokine CXCL14（BRAK） induced the migration of human renal cell carcinoma, A-498 cells
摘要
腎細胞癌（renal cell carcinoma；RCC）是腎臟惡性腫瘤中最常見的疾病，主要源於近端腎小管細胞。腎癌初期的症狀並不明顯，多數的病人發現時已經處於腎癌末期並伴隨著強烈的抗藥性，且多半患者已具有轉移的現象。腎細胞癌發生轉移後的病人存活率相當低，因此找出RCC的分子標記以利於RCC的治療是目前重要的課題。趨化激素（Chemkine）在早期的研究主要是與免疫和發炎反應有關，近年來有越來越多研究發現趨化激素與腫瘤的轉移和血管新生扮演著很重要的角色。先前有文獻指出CXCL14扮演調控胰臟癌細胞、前列腺癌細胞以及乳癌細胞轉移（migration）和侵犯（invasion）的功能。根據先前我們實驗室cDNA microarray的分析得知CXCL14在腎癌細胞中有大量表現的情形，因此我們推測CXCL14可能與癌症的惡化有關。在本研究首先利用real-time PCR，西方點墨法檢測腎癌細胞CXCL14基因的表現，我們也利用Enzyme-linked immunosorbent assay （ELISA）檢測腎癌細胞分泌CXCL14的情形，發現CXCL14在腎細胞癌中的蛋白質表現和分泌量都比正常的腎細胞株具還較高，接著外加處理CXCL14進行細胞移行的實驗，結果發現CXCL14確實具有刺激腎癌細胞移行的能力。進一步利用西方點墨法探討其分子機制，發現CXCL14透過增加EGFR與Integrin表現增強，Erk、Src、AKT訊號活化，在細胞遷移過程中相關的Rho GTPases家族分子如：RhoA、Rac、Cdc42也觀察到表現有增加的情形，因此我們推測CXCL14在腎細胞癌中扮演促進細胞轉移的角色，也可能可以當作腫瘤轉移的分子標記。
並列摘要
Renal cell carcinoma (RCC) is one of the most common kidney diseases and it is mainly developing from proximal tubular cells. The early symptoms of kidney cancer are not obvious; in most cases they were already the last phases when patients discovered them, and there had also appeared metastatic phenomenon. Survival rates of patients with spread RCC are very low, therefore, it is a very important issue to identify molecular markers of RCC in order to improve RCC therapy. In most earlier studies, chemokine is mainly related to immune and inflammatory responses, however in recent years there are more and more studies have realized the important role chemokine plays in tumor metastasis and angiogenesis. Some previous documents have indicated that, CXCL14 plays various functions such as regulation of pancreatic cancer cells, prostate cancer cells; migration and invasion of breast cancer cells. According to our previous analysis of cDNA microarray done in laboratory, we know that CXCL14 is massively expressing in renal cancer cells, thence, we suppose CXCL 14 is related to deteriorating cancers. In this study, we first adopted real-time PCR and Western blot to detect expressions of CXCL14 gene in renal cancer cells and we also used Enzyme-linked immunosorbent assay (ELISA) to detect the status how renal cancer cells secreted CXCL14.We found the values of protein expressions and secretions were way higher than normal kidney cells. Afterward, we processed experiments for CXCL14 migrations and we found CXCL14 exactly had abilities to stimulate migrations of renal cancer cells. Further, we still adopted Western Blot to explore its molecular mechanisms and we found CXCL14 was strengthen by increasing EGFR and Integrin expressions and activated signals of Erk, Src and AKT. Among relevant family molecules during cell migrations such as RhoA, Rac and Cdc42, we also observed increases of expressions. Therefore, we concluded CXCL14 to be the role of promoting cell metastasis in renal cell carcinoma and it could also be used as molecular markers for tumor metastasis.
參考文獻
延伸閱讀
- 劉斐翠（2009）。趨化激素 CXCL14 (BRAK) 在人類腎細胞癌扮演之分子角色〔碩士論文，高雄醫學大學〕。華藝線上圖書館。https://www.airitilibrary.com/Article/Detail?DocID=U0011-2707200913004300
- 陳彥羽（2019）。探討知母皂苷AIII在腎臟癌細胞轉移之扮演角色和分子機轉〔碩士論文，中山醫學大學〕。華藝線上圖書館。https://doi.org/10.6834/CSMU201900148
- 何佳珞（2014）。探討SPZ1引起人類腎細胞癌化學抗藥性與EGFR活化之關係〔碩士論文，高雄醫學大學〕。華藝線上圖書館。https://www.airitilibrary.com/Article/Detail?DocID=U0011-2508201415445200
- 孫瑛穗（2008）。乳薊萃取物(silibinin)抑制腎臟癌細胞侵襲和轉移的能力及輔助抗癌藥(taxol, vinblastine, 5-fluorouracil)誘導細胞凋亡機制之探討〔碩士論文，中山醫學大學〕。華藝線上圖書館。https://doi.org/10.6834/CSMU.2008.00019
- 蘇士豪（2011）。Molecular mechanism of SPZ1 mediated drug-resistance in human renal cell carcinoma〔碩士論文，高雄醫學大學〕。華藝線上圖書館。https://www.airitilibrary.com/Article/Detail?DocID=U0011-2908201119303400