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研究生:余乾碩
研究生(外文):CHIEN-SHUO YU
論文名稱:蒙古黃耆對大腸癌影響並降低miR-29a的表現量之研究
論文名稱(外文):Down regulation of miR-29a by Astragalus mongholicus treatment in Colorectal Cancer
指導教授:蘇立仁
指導教授(外文):LI-JEN SU
學位類別:碩士
校院名稱:國立中央大學
系所名稱:系統生物與生物資訊研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2019
畢業學年度:107
語文別:中文
論文頁數:60
中文關鍵詞:黃耆大腸癌降低表現量研究
外文關鍵詞:Down regulationmiR-29aAstragalus mongholicusColorectal Cancer
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蒙古黃耆 (學名:Astragalus mongholicus,AM) ,在過去數篇文章中指出將它用於治療癌症有一定的功效[1-4],但其中的分子機轉的部分仍然尚未明確。本研究目的是欲找出黃耆加藥後影響大腸癌細胞以及組織的分子機轉,用系統生物的分析技術去研究黃是是如何影響或治療大腸癌。本研究先從基因晶片分析開始,使用GeneSpring軟件GX 7.3版(安捷倫,CA,USA)先行選出差異基因,並使用miRTarBase生物資料庫合併分析,最後再結合曾愛倫博士的論文內容合併篩選出候選的基因---的miR-29a[5],一個非編碼RNA,以進行後續的實驗。本研究使用即時聚合酶鏈鎖反應實驗先去證明在大腸癌HCT116細胞株以及使用本研究室過去研究生曾愛倫留下來的24隻小鼠組織樣本,這些組織樣本是先將牠們皮下注入HCT116細胞並長出腫瘤,再接受黃耆加藥治療,最後在將腫瘤組織處理研究。組織中的miR-29a的表現量有因黃耆造成出現抑制的影響。之後,由於本研究很榮幸能跟陳文逸老師實驗室有合作,因此本研究得以獲得陳老師的專利nDNA修飾的探針使用的權利,以用於進行原位雜交實驗。我們可以確認的miR-29a在HCT116細胞株中表現量有因黃耆加藥後而產生差異。之後本實驗使用先前本研究室留下的小鼠組織陣列晶片去進行組織學原位雜交實驗,進一步去證明黃耆加藥後,在組織學上亦可以看出黃耆加藥之後造成的miR-29a表現量的抑制。或許在未來的miR-29a的標靶藥物會結合黃耆開發出新的醫學以及商業用途用來治療大腸癌。
Astragalus Mongholicus (AM), has pointed out in the past several articles that it has effect on the treatment of colorectal cancer, but the molecular mechanism of the part is still unclear. The purpose of this study is to find out the molecular mechanism of the influence of jaundice on colorectal cancer cells and tissues, and to study how Astragalus mongholicus affects or treats colorectal cancer from the perspective of molecular biology. This study begins with the analysis of the gene chip, uses the GeneSpring software GX version 7.3 (Agilent, CA, USA) to select the differential genes first, and uses the miRTarBase biological database to combine the analysis, and finally combines the previous research results of the laboratory to select the candidate genes. --- miR-29a, a non-coding RNA for subsequent experiments. In this study, the real-time polymerase chain reaction assay was used to prove that the HCT116 cell line of colorectal cancer and the 24 Rats left by the graduate students Zeng Heather in this laboratory were injected subcutaneously into HCT116 cells and grew tumors, and then received jaundice. Drug treatment, and finally in the treatment of tumor tissue. The amount of miR-29a in the tissue is affected by the inhibition caused by jaundice. Later, because this study has cooperated with Chen Wen yih laboratory, this study obtained the right to use Chen's patented nDNA modified probe for in situ hybridization experiments. We can confirm that the expression of miR-29a in HCT116 cell line is different due to the addition of jaundice. After this experiment, the mouse tissue array wafer left by the previous laboratory was used for histological in situ hybridization experiment, and further proved that after the administration of jaundice, the miR caused by the administration of jaundice can also be seen in histology. -29a inhibition of the amount of expression. Perhaps in the future, miR-29a's target drugs will be combined with Astragalus to develop new medical and commercial uses for the treatment of colorectal cancer.
目錄
中文摘要 I
Abstract II
致謝 III
圖目錄 V
第1章介紹 1
1.1流行病學 1
1.2治療 2
1.3結直腸癌的分子分析 2
1.4意義和目標 4
第二章材料和方法 5
2.1細胞和培養條件 5
2.2 alamarBlue®生存力分析 5
2.3 FACS分析 5
2.4微陣列基因表達晶片分析 6
2.5即時聚合酶鏈式反應實驗 6
2.6具有差異表達的微小RNA的數據庫分析 7
2.7原位雜交實驗 7
第3章黃耆影響大腸癌細胞內的微小RNA的研究結果 9
3.1介紹和目標 9
3.2結果 11
3.2.1 黃耆抑制體外HCT116細胞的生長 11
3.2.2 miRNA的分析 12
3.2.3 即時聚合酶鏈鎖反應實驗 17
3.2.4細胞學及組織學原位雜交實驗 18
3.2.5功能分類、途徑和網絡分析 22
第4章結論 27
第5章討論 28
參考文獻 30
參考資料 35
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