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Title: | 利用化學蛋白體學鑑定hnRNP A1為槲黃素之目標蛋白與其對攝護腺腫瘤細胞之影響探討 Chemical proteomics reveals hnRNPA1 as a quercetin target and its effects in PC-3 cell |
Authors: | Yun-Ju Chen 陳韻如 |
Advisor: | 周綠蘋(Lu-Ping Chow) |
Keyword: | 攝護腺癌,化學蛋白體學,槲黃素,hnRNP A1, prostate cancer,chemical proteomics,quercetin,hnRNP A1, |
Publication Year : | 2011 |
Degree: | 碩士 |
Abstract: | 檞黃素是一種普遍存在於蔬果和穀物中的植物性生物黃酮,已經被用在治療慢性攝護腺炎病患。槲黃素是天然的抗組織胺、抗發炎劑、抗氧化劑,還具有抗癌的功效。據研究報導,槲黃素可以誘導攝護腺癌細胞凋亡和抑制癌細胞的生長,並且是具有選擇性的殺死癌細胞而不影響到正常細胞。我們已經從文獻知道槲黃素會影響許多機制,但我們想更進一步了解是否還有我們尚未發現的機制與未知的槲黃素標靶蛋白,使得槲黃素能夠阻礙腫瘤細胞的生長。我們應用了最近新興的化學蛋白體學技術去找尋在攝護腺癌細胞中會與槲黃素交互作用的蛋白。化學蛋白體學是結合了傳統藥物親和層析和質譜儀的生物資訊分析技術以鑑定蛋白。在我們的研究結果中,我們發現hnRNP A1與槲黃素有交互作用,並在後續的體外實驗,以免疫沉澱、西方墨點法、和濾紙結合實驗確定了hnRNP A1和槲黃素之間直接的結合。我們的研究指出,槲黃素與hnRNP A1的結合會阻礙hnRNP A1在細胞核質間往返的能力,造成hnRNP A1堆積在細胞質中,進而阻礙mRNA進行轉譯,影響了抗凋亡蛋白的表現。隨後,細胞內蛋白的不平衡及抗凋亡蛋白的減少則造成癌細胞的死亡。我們對於「槲黃素-hnRNP A1-攝護腺癌細胞」提出新的機制,發現槲黃素與hnRNP A1之間的關連,並更加清楚了解槲黃素分子的作用機轉與可能的副作用。hnRNP A1以後可能可以作為新的藥物標靶蛋白,以用來治療復發性的非男性荷爾蒙依賴型攝護腺癌。同時,我們建立起化學蛋白體學技術平台,可以應用到各種藥物分子和疾病上,幫助了解藥物分子機制及新藥物的開發。 Quercetin is a plant-derived flavonoid found in fruits, vegetables, leaves and grains, and it is one of the pharmaceutical molecules used to treat patients with chronic prostatitis. Quercetin had been reported that it could induce cell apoptosis and inhibit cell growth in PC-3 cells and also enhance its anti-cancer efficacy when combining other dietary phytoestrogens. It appears to be a selective inhibitor of cell proliferation of tumor cells instead of normal cells. There were many mechanisms of quercetin involved, but we want to know if there still are some new pathways to hamper tumor progression that we did not know before. In order to find out the direct molecular targets of quercetin in its medicinal property in prostate cancers, we applied chemical proteomics techniques to reach our goal. Chemical proteomics is a postgenomic version of classical drug affinity chromatography that is coupled to subsequent high-resolution MS and bioinformatic analyses for protein identifications. We identified hnRNP A1 by using quercetin-affinity column and demonstrated a specific interaction between quercetin and hnRNPA1 by immunoprecipitation, immunoblotting and in vitro binding experiments. There were many studies indicated that aberrant mRNA processing, nuclear export, splicing and translation would cause imbalance of protein homeostasis and high risk of cancer occurring. Our studies pointed out the interaction of hnRNPA1 and quercetin impairs hnRNPA1’s nucleocytoplasmic shuttling and having impacts on the translation of mRNAs related to cell anti-apoptosis and cell proliferation. We connect the link of quercetin- hnRNPA1- prostate cancer, and try to clarify the mechanism of quercetin treatment to PC-3 cells and finally tumor growth decreased. Our study is a promising example and powerful platform to elucidate a potential drug’s target proteins and might apply to further drug designs to strength the specificity and selectivity. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/36185 |
Fulltext Rights: | 有償授權 |
Appears in Collections: | 生物化學暨分子生物學科研究所 |
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