博碩士論文 105826002 詳細資訊




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姓名 蕭如晴(Ju-Ching Hsiao)  查詢紙本館藏   畢業系所 系統生物與生物資訊研究所
論文名稱 透明腎臟細胞癌發生前期與組織發炎之關係研究
(Study of pre-cancerous tissue inflammation in the development of clear-cell renal cell carcinoma)
相關論文
★ 由基因微陣列分析發炎與腎臟細胞癌發生之機制★ VHL基因突變在癌前期的組織發炎機制
★ VHL剔除模型之轉錄體差異以及台灣透明細胞腎細胞癌族群之特定基因體變異之研究★ VHL knockdown HK-2 cells induce macrophage endothelial extravasation
★ ITPR2, an ER calcium channel, regulates ER stress and inflammatory response in pre-cancerous kidney tubule cells★ 應用大腸桿菌與酵母菌蛋白質體晶片系統性分析抗菌肽及抗生素作用之目標蛋白質
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摘要(中) 馮·希佩爾·林道(VHL)基因是抑癌基因。 VHL 基因的缺失會導致人類透明細胞腎細胞癌(ccRCC)的發生。在先前的研究中,我們證明了腎小管細胞中剔除小鼠VHL 等位基因(Vhlh)會導致組織炎症和增生。但是ccRCC 和炎症之間的機制仍然不清楚。
另一方面,過去的研究表明他汀類藥物具有抗發炎作用。VHL 基因剔除小鼠會產生腎臟炎症的病理徵象,並可能進一步引起腎臟細胞的病理變化,例如ccRCC。因此,我們對VHL 基因剔除小鼠進行他汀類抗炎藥物注射試驗,通過已知的藥物對影響癌症的炎症機制基因治療實驗以進一步探討他汀類藥物對VHL 突變小鼠的腎臟炎症相關反應。
在這項研究中,我們也結合了顯性突變p53R172H / + 和Vhlh 基因剔除雙突變的表達,將其作為ccRCC 的新型模型。我們的結果表明,Ki-67 蛋白在p53R172H / + 和Vhlh 雙等位基因組織中具有高度表達,並且在上皮細胞中透明細胞堆積及囊泡變得明顯且增多。結果表明,p53R172H /+ 和Vhlh 雙等位基因突變可作為研究炎症誘導和ccRCC 形成機制的新小鼠模型。
摘要(英) The von Hippel-Lindau (VHL) gene is a tumor suppressor gene. Loss of the VHL gene causes clear-cell renal cell carcinoma (ccRCC) in human. In previous study, we demonstrated that knockout of the mouse VHL allele (Vhlh) in the kidney tubule cells resulted in tissue inflammation and hyperplasia. However, the relationship between ccRCC and inflammation remains unclear.
In order to further test whether tissue inflammation is important for hyperplastic transformation, we examined the efficacy of statins in relieving the fibrotic and proliferative phenotypes of the Vhlh knockout kidney, since past studies have shown that statins have anti-inflammatory effects. Vhlh knockout mice produce pathological signs of kidney inflammation and may further cause changes in kidney cells that lead to ccRCC. Therefore, we performed an anti-inflammatory statin injection test on Vhlh knockout mice to examine the causal relationship between inflammation and hyperplasia. The result indicates that statin may be a potential preventive agent for blocking tumor development in inflamed tissue.
In this study, we also combined the expression of dominant-negative p53R172H/+ and Vhlh knockout mutations as a novel model of ccRCC. Our results indicated that proliferation marker Ki-67 protein is highly expressed in p53R172H/+ and Vhlh double allele tissue, and cystic and pile-up epithelia became apparent. The results suggest that p53R172H/+ and Vhlh double allele could be used as a novel model in the study of mechanism underlying inflammation-induced ccRCC formation.
關鍵字(中) ★ 腎透明細胞癌
★ 發炎機制
★ 他汀類藥物
關鍵字(英) ★ ccRCC
★ VHL
★ p53
★ Simvastatin
論文目次 中文摘要.........................................................................................................................I
Abstract..........................................................................................................................II
致謝...............................................................................................................................III
Chapter 1 Introduction ...................................................................................................1
1-1 kidney cancer and the VHL gene .....................................................................1
1-1-1 Kidney ..................................................................................................1
1-1-2 ccRCC ..................................................................................................1
1-1-3 The function of VHL gene ....................................................................3
1-1-4 Inflammation and cancer .................................................................5
1-1-5 Anti-inflammation effect of the statin drug..........................................7
1-1-6 Vhlh and simvastatin ............................................................................8
1-1-7 p53........................................................................................................8
1-1-8 Dominant negative mutation ................................................................9
1-1-9 p53R172H/+ dominant negative mutation..............................................10
Chapter 2 Materials and Method..................................................................................12
2-1 Experimental materials..................................................................................12
2-1-1 p53 and Vhlh mutant mouse model....................................................12
2-1-2 Phosphate buffer saline solution (PBS)..............................................13
2-1-3 Sirius Red Solution and staining ........................................................13
2-1-4 P53R172H/+ Allele .................................................................................14
2-2 Method...........................................................................................................15
2-2-1 Animal model of Vhlhloxp/loxp allele ....................................................15
2-2-2 preparation of simvastatin ..................................................................15
2-2-3 Simvastatin IP injection......................................................................15
2-2-4 Generate Vhlh-p53R172 H/+ double mutant...........................................16
2-2-5 IHC staining (Ki-67) ..........................................................................17
2-2-6 IHC staining (Sirius Red)...................................................................18
2-2-7 Tissue Analysis...................................................................................19
Chapter 3 Results and Discussion................................................................................20
3-1 Kidney tissue H&E staining of Vhlh mutant mouse......................................20
3-2 Simvastatin IP injection reduces inflammation .............................................22
3-2-1 Simvastatin drug IP injection of wile type (WT) control...................23
3-3-2 Simvastatin drug IP injection rescued phenotypes of Vhlh mutant mice
......................................................................................................................24
3-3 Quantification of fibrotic index (sirius red staining).....................................25
3-4 Relationship between simvastatin and clear cell formation ..................25
3-5-1 mouse breeding ..................................................................................30
3-5-2 Characterization of Vhlh and p53R172H/+ double mutation..................31
3-5-4 p53 mutant enhances proliferation index in Vhlh knockout...............33
3-5-6 Vhlh/p53
R172H/+
double allele exhibits hyperplastic growth ...............35
Chapter 4 Conclusion...................................................................................................36
Reference .....................................................................................................................39
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指導教授 徐沺(Tien Hsu) 審核日期 2020-8-20
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