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博碩士論文 etd-0720113-005056 詳細資訊
Title page for etd-0720113-005056
論文名稱
Title
泛素接合酶UBE2C 在乳房癌變中的角色
The Role of Ubiquitin-Conjugating Enzyme UBE2C in Breast carcinogenesis
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
73
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2013-06-28
繳交日期
Date of Submission
2013-08-20
關鍵字
Keywords
乳癌、乳房微鈣化、UBE2C (ubiquitin conjugation E2 enzyme)、人類乳癌細胞
Breast cancer, Breast microcalcification, UBE2C (ubiquitin conjugation E2 enzyme), human breast cancer cells
統計
Statistics
本論文已被瀏覽 5702 次,被下載 476
The thesis/dissertation has been browsed 5702 times, has been downloaded 476 times.
中文摘要
乳房微鈣化組織為乳癌發生初期的一個重要特徵,可透過乳房 X光攝影進行診斷。乳房微鈣化點可來自乳房發炎、管內乳頭狀瘤、纖維腺瘤、纖維囊腫、脂肪細胞壞死所產生的鈣化和乳癌等。約 85% 的乳房組織微鈣化為良性,只有 15% 的鈣化可能合併癌症,由於鈣化本身可以是一種正常或不正常細胞死亡代謝生理現象或結果,因此乳房微鈣化組織並不就是等同於乳癌。然而若能及早偵測出乳房微鈣化組織,並分析這些微鈣化的生化特性,可為有效防治乳癌的方法。 UBE2C 為泛素蛋白解體系統(ubiquitin-proteasome system, UPS)中的一種 E2 泛素接合酶(E2 conjugating enzyme),主要參與細胞生長週期的運行。研究發現,在多種癌組織中可檢出 UBE2C 的大量表達,因此 UBE2C 被鑑別為一種新的腫瘤生物標記。由於 UBE2C 在乳癌形成中所扮演的角色仍不清楚,此研究針對 UBE2C 的表現與乳癌之間的關係進行分子生化與細胞生物學的分析,並結合臨床病理學之診斷報告以評估 UBE2C 在臨床應用的可能性。此研究透過乳癌篩選所納入之臨床採樣人數共計 72 位,其中完成 RT-qPCR 分析者計 56 位,完成免疫組織化學染色者計 50位。乳房微鈣化組織檢體及未微鈣化檢體以 RT-qPCR 實驗分析 UBE2C mRNA 的表現,結果顯示約有 70% 的微鈣化組織檢體其 UBE2C 表現量比未微鈣化組織增加 2 倍以上,我們將 UBE2C mRNA倍數增加之數據與病理學診斷結果分析比對,發現 UBE2C 的表現量與乳房組織之癌化的具有關聯性。另外免疫組織化學染色結果顯示約有 68% 的微鈣化組織病理切片呈現 UBE2C 陽性。為進一步探討 UBE2C 在乳癌發生的過程中的分子調控機制,我們分析三株乳癌細胞株 MCF-7,MDA-MB-231 及 MDA-MB-361 的 UBE2C 表達情形,其結果顯示具侵襲性的乳癌細胞 MDA-MB-231 及 MDA-MB-361 高度表達 UBE2C,而不具侵襲性之乳癌細胞 MCF-7 表現的 UBE2C 則相對較低。分析shRNA 抑制UBE2C 表現的 MCF-7 細胞,發現其細胞的生長複製被抑制,而且細胞凋亡的程度增加;反之,過度表現外源性 UBE2C (overexpression)則提升了 MCF-7 細胞增生的能力。我們的結果證實 UBE2C 在乳癌細胞增生與癌化過程中扮演重要角色。
Abstract
Breast microcalcification is an important feature of early breast carcinoma. Mammography and histology diagnosis reveal that 15% of breast microcalcification could be further defined as breast cancer. UBE2C, an ubiquitin conjugation E2C, is a newly identified biomarker candidate in various types of cancers. It contributes to the ubiquitin-mediated proteasome degradation in cell cycle. To evaluate the clinical implication of UBE2C in early breast cancer diagnosis, we measured the mRNA expression level of UBE2C from 56 sets of breast biopsy samples; each set contains non-microcalcification and microcalcification tissues from individual. Approximately 70% microcalcification samples with 2 fold increased UBE2C mRNA expression were detected by quantitative RT-PCR; in addiction, the immunohistochemistry also revealed that 68% microcalcification tissues were UBE2C positive. The UBE2C expression level and pathology diagnosis data of the microcalcification biopsies were future analysed. The results indicated a correlation between highly expression of UBE2C and early breast cancer, which suggested that UBE2C might be a comportable biomarker of early breast cancer. To further investigate the role of UBE2C in breast cancer oncogenesis, we determined the UBE2C expression level in 3 types of breast cancer cells. We found that the invasive cells, MDA-MB-231 and MDA-MB-361, highly express UBE2C, and the relative lower expression of UBE2C were observed in non-invasive cells, MCF-7. By using sh-RNA gene silencing vector of UBE2C, the cell growth reduction and cell death increasing were found in UBE2C knockdown MCF7 cells. In contrast, the MCF-7 growth rate was enhanced by over-expression of UBE2C. Our data suggest that the UBE2C plays the important role in promoting cell proliferation and breast oncogenesis.
目次 Table of Contents
中文摘要 iii
英文摘要 vi
目錄 viii
圖表目錄 ix
壹、導論 1
一、乳癌 1
二、乳房微鈣化 3
三、UBE2C 4
貳、研究動機 7
參、材料與方法 8
肆、結果 26
伍、討論 33
陸、圖表與說明 37
柒、參考文獻 51
捌、附錄 55
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