肝癌(Hepatocellular carcinoma, HCC)為全球性常見的惡性腫瘤之一，儘管是初期之肝癌在治療上也是有相當的挑戰。雖然已有許多研究找出與肝癌相關的生物標記，但也不難在肝癌病患中發現這些常見生物標記有低表現的情形。因此研究於結合肝癌有力的生物標記而達早期診斷的目的為相當重要的。為此我們在先前的實驗中於互補去氧核糖核酸資料庫裡篩選出在肝癌有著高度表現的CDC28 protein kinase regulatory subunit 1B (CKS1B) 基因，並且在臨床的結果發現了有預後不良的情形發生在高表現CKS1B的病患中。此外在一些文獻指出了在肝癌中細胞內染色體其中的1q為變異最大的染色體，而1q21為最高度擴增的區域，哺乳類的CKS1B基因就是座落於此高度擴增區域1q21.2-q22，並在演化上其胺基酸序列為相當高度保守。CKS1B在一些癌症文獻中被報導為扮演著致癌的角色，而目前最熟知的路徑為，CKS1B會做為S-phase kinase-associated protein 2 (SKP2) 的輔因子與SKP2為E3 ligase所組成的SKP1-Cullin1-F-box泛素化複合體進而影響細胞週期抑制蛋白cyclin-dependent kinases 1A (p27kip1) 的降解，而無法正常抑制細胞週期的細胞將會造成不正常的增生。本研究結果顯示在外源性轉染CKS1B表現質體的肝癌細胞株中明顯地造成了其增殖與侵犯性的上升，相對的以short hairpin RNA干擾技術發現在抑制CKS1B的表現後使其存活與增殖率下降，並且會增加肝癌細胞的凋亡，這些結果說明了CKS1B在肝癌生成中有著多種的功能扮演著重要角色。

Hepatocellular carcinoma (HCC) or hepatoma is the fifth most common cancer and the third most common cause of cancer-related deaths worldwide. Early detection of HCC still remains challenge. Patients with late-stage HCCs have very limited therapeutic options and relatively poor prognosis. Therefore, recognition of useful tumor marker for the diagnosis of HCC may aid in the management with this lethal malignancy, and designing effective therapies.
In our previous study, through in silico data mining of numerous available cDNA microarray databases, the CDC28 protein kinase regulatory subunit 1B (CKS1B) transcript was found to be frequently upregulated in HCCs and the gene is strongly associated in clinical aggressiveness. Moreover, gain of chromosome 1q copy is one of the most frequently detected alterations in HCC and 1q21 is the most frequent minimal amplifying region. Mammalian CKS1B gene, a member of the highly conserved cyclin kinase subunit 1 protein family, mapped to 1q21.2-q22, which interacts with cyclin-dependent kinases and frequently up-regulated in human HCC specimens, and this gene also has been reported to play oncogenic roles in other human cancers. Furthermore, this gene was identified as an essential cofactor of S-phase kinase-associated protein 2 (SKP2) in the SKP2-containing SKP1-Cullin1-F-box ubiquitin ligase complex–mediated ubiquitination of the cell cycle inhibitor protein cyclin-dependent kinases 1A (p27kip1) that leads to the proteasomal degradation of p27kip1. In the present study, our results indicate that CKS1B overexpression may contribute to hepatocarcinogenesis by enhancing the cellular tumorigenic potential. In addition, CKS1B was correlated with the HCC-derived cells proliferation, and CKS1B knockdown significantly inhibited the tumorigenic potential of HCC-derived cells. In this study, we examined that CKS1B knockdown and overexpressed in HCC cells significantly changed the expression of genes known to participate in various cellular processes, including, apoptosis, cell cycle, proliferation, viability, invasion, and migration, implying that CKS1B may play considerable and diverse roles in hepatocarcinogenesis.

論文審定書..........................................................................i
致謝.....................................................................................ii
摘要.....................................................................................iii
Abstract………………………………………….....................iv
Introduction...……………………….....................................1
Materials and methods……................................................4
Results...............................................................................14
Discussion....……………………….....................................29
References.....………………………...................................31
Appendix.….......................................................................34

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