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| 題名: | 探討CCNE2 (Cyclin E2)在乳癌中所扮演的角色 |
| 作者: | 劉明芳 |
| 貢獻者: | 醫學科學研究所 |
| 日期: | 2010 |
| 上傳時間: | 2010-10-20 11:50:58 (UTC+8) |
| 摘要: | 根據行政院衛生署生命統計數據顯示，國人近十年來女性乳癌發生率及死亡率均有逐年攀升情形，且有年輕化趨勢，因此，探討乳癌相關研究也就刻不容緩。
本研究主要是探討Cyclin E2與乳癌細胞、臨床病患相關性；透過細胞株與臨床檢體實驗，觀察Cyclin E2在乳癌中所扮演的角色。在細胞實驗上，藉由反轉錄-聚合酶連鎖反應方法，初步觀察Cyclin E2在細胞株表現情形，進而選擇實驗室常用MDA-MB-231、MCF-7乳癌細胞株作天然化合物木犀草素及數種人工化合物24小時處理，發現在乳癌細胞株MDA-MB-231有明顯抑制細胞生長現象，且Cyclin E2 mRNA/Protein表現量會隨著劑量增加而減弱。在臨床方面，則利用即時定量聚合酶連鎖反應方法去偵測257名乳癌病患之正常組織及癌化組織檢體之Cyclin E2表現量，同時收集病患診斷相關資料，並應用SPSS統計軟體作分析，進而發現Cyclin E2在癌化組織有較高表現量；且正常組織與癌化組織表現量比值超過10倍者，在T＞N組病例發生率是較高。而在臨床乳癌病患之病理組織切片利用免疫組織化學染色方法，亦發現Cyclin E2在乳癌組織癌細胞區域呈現陽性棕色反應。
According to the Department of Health, Executive Yuan,R.O.C. vital statistics data show that people over the past decade the incidence of female breast cancer and mortality cases are increasing year by year, and there is getting younger and younger, therefore, studies of breast cancer also imperative.
This study was to Cyclin E2 and breast cancer cells, clinical disease associated; through cell experiments and clinical samples, observe Cyclin E2 in the role of breast cancer. In cell experiments, by reverse transcription - polymerase chain reaction method, preliminary observations Cyclin E2 expression in cell lines and select the laboratory used MDA-MB-231, MCF-7 breast cancer cells to natural compounds luteolin and artificial compounds treatment 24 Hrs, found that breast cancer cell line MDA-MB-231 significantly inhibited the growth of the phenomenon, and Cyclin E2 mRNA / Protein expression levels decreased as the dose increases. In clinical terms, the use of real-time quantitative polymerase chain reaction method to detect breast cancer patients of 257 normal tissue and cancerous tissue samples of Cyclin E2 expression, and to collect patient diagnostic information, application of SPSS statistical software for analysis , Cyclin E2 was found in cancerous tissue had higher expression; and normal tissue and cancerous tissue of more than 10 times the performance compared to those in the T＞N group is a higher incidence of cases. Immunohistochemical stain of CCNE2 expression in clinical patient tissues, we observed that Cyclin E2 in tumor region of breast cancer tissue showed positive reaction(brown). |
| 關聯: | 80頁 |
| 描述: | 中文摘要(Abstract in Chinese) I~II
英文摘要(Abstract in English) III~IV
目錄(Contents) V~VII
圖目錄(Figure lists) VIII
縮寫表(Abbreviations) IX ~XI
第一章緒論(Introduction) 1
前言 2
一、乳癌簡介 2
二、Cyclin E2 6
三、天然化合物木犀草素(Luteolin) 7
第二章研究目的與實驗架構 (Purposes and construction) 9
第三章實驗材料與方法 (Materials and Methods) 11
一、藥品與試劑 12
二、常用溶液 15
三、常用儀器 16
四、實驗方法 18
(一)Primer設計及配製 18
(二)細胞培養及藥物處理 19
(三)mRNA萃取 21
(四)反轉錄-聚合酶連鎖反應及核酸洋菜凝膠電泳 22
(五)即時定量聚合酶連鎖反應 25
(六)蛋白質聚丙烯醯胺凝膠電泳及西方式墨點法分析 27
(七)免疫組織化學染色 29
(八)細胞存活率分析 31
第四章實驗結果 (Results) 32
一、Cyclin E2在乳腺細胞株中mRNA的表現 33
二、Cyclin E2在乳腺細胞株中Protein的表現 33
三、臨床乳癌病患之正常組織及癌化組織檢體，利用Real-time PCR偵測Cyclin E2 mRNA，發現在癌化組織中有較高表現量 34
四、臨床乳癌病患之正常組織及癌化組織檢體，透過Real-time PCR方法所偵測到Cyclin E2 copy number，進一步與臨床醫師所提供病患診斷資料，經SPSS統計軟體分析其相關性 35
五、臨床乳癌病患之病理組織切片利用免疫組織化學染色方法，發現Cyclin E2在乳癌組織癌細胞區域被偵測(棕色) 36
六、天然化合物Luteolin對乳癌細胞株MDA-MB-231及MCF-7之Cyclin E2的表現中，發現在MDA-MB-231細胞Cyclin E2表現被抑制 37
七、天然化合物Luteolin對乳癌細胞株MDA-MB-231及MCF-7細胞生長的影響 38
八、數種人工化合物具有抑制乳癌細胞株MDA-MB-231之Cyclin E2的表現 39
第五章討論 (Discussion) 40
第六章圖表 (Figures) 44
第七章參考文獻 (References) 62
附錄 (Appendices) 71
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