請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26390完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 郭彥彬,蕭宏昇 | |
| dc.contributor.author | Meng-Yi Lin | en |
| dc.contributor.author | 林孟誼 | zh_TW |
| dc.date.accessioned | 2021-06-08T07:08:34Z | - |
| dc.date.copyright | 2008-09-11 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-08-01 | |
| dc.identifier.citation | References
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26390 | - |
| dc.description.abstract | 我們發展了一種方便的基因轉染進入細胞的方式(surfection)，將DNA PEI Gelatin複合物混合乾燥在96 well細胞培養盤的表面並儲存在-80℃直到將細胞放入進行基因轉染。跟一般的基因轉染方式比較， surfection 可以使用在有血清的狀況下，重要的是，DNA/PEI 複合物幫助細胞黏貼在培養盤上。而且，培養盤經過長時間的儲存後並不會降低轉染效率也不會對細胞有毒性。因為此複合物的的穩定性，surfection可以用來進行大規模的基因轉染。對於蛋白質，基因調控和病毒產生的篩選都可以使用。而且有再現性、價格低、多方面適用在不同的應用上。
在台灣，頰黏膜鱗狀細胞癌佔男性惡性腫瘤的發生率的第四位。發病原因和抽煙以及吃檳榔等生活習慣息息相關。已經有研究證實尼古丁和鹼性環境會誘發磷酸化Akt作用和口腔癌的形有關。 魚藤素(deguelin)是一種類黃酮家族中的類血藤酮，可以經由抑制Akt來降低在動物模式中在肺臟、腸、乳房和皮膚中腫瘤的發生。我們使用surfection去生產慢病毒的shRNA進而感染口腔癌細胞SAS。在口腔癌中進行高速RNAi篩選分析Akt 路徑對細胞生長或死亡的調控。結合魚藤素(deguelin)和RNAi進行高速篩選在分析化合物的作用機制和未知但想研究的副作用以及識別對於藥物敏感有潛力的基因標靶有特別的優勢。 | zh_TW |
| dc.description.abstract | We developed a straightforward surfection method for cells wherein DNA/polyethylenimine/gelatin complexes are coated on the surface of 96 well plates and stored in -80℃ until cells are ready for transfection. Comparing to most of the conventional transfection, surfection can be performed in the presence of serum. Importantly, DNA/polyethylenimine complexes promote cell adhesion to the plates. Also, long-term storage of the plates did not reduce the transfection efficiency nor it had any effects on the cell toxicity. Because of the stability of complexes, surfection enables large-scale tansfection thus providing a reproducible, cost-effective and versatile tool for parallel screening of proteins, gene regulatory elements and virus production used in diverse applications.
In Taiwan, the incidence and the mortality of oral squamous cell carcinoma (OSCC) were ranked fourth in men among all malignancies. It is well documented that cigarette smoking and areca nut chewing contribute to the risk of oral squamous cell carcinoma (OSCC). The role of phosphorylated Akt (p-Akt) in oral carcinogenesis induced by nicotin and alkaline environments was investigated. Deguelin, a rotenoid of the flavonoid family, is able to decrease tumor incidence in animal models for lung, colon, mammary and skin carcinogenesis through Akt inhibition. We used surfection to produce lentivirus shRNA to infect oral cancer cell SAS. High-throughput RNAi screening along Akt pathway was carried out in oral cancer cell to analyze cell apoptosis and growth. High throughput screening with Degulin and RNAi is of particular value as it’s not only for analyzing a compound’s mechanism of action and understanding of unwanted side effects but also for identifying potential gene targets for developing sensitizing agents for existing drugs. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T07:08:34Z (GMT). No. of bitstreams: 1 ntu-97-R95450003-1.pdf: 4085922 bytes, checksum: 560d1a7e5e45598e8ef2a9896707c8ad (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | Contents 1
Abbreviations 2 中文摘要 3 Abstract 4 Chapter 1 General Introduction 6 1.1 Background and significance 7 1.2 Specific aims 20 Chapter 2 Materials and Methods 21 Chapter 3 Results 30 3.1 To determine adaptable volume of plasmid in surfection 3.2 To study the applicability of surfection 3.3 To determine the stability of surfection 3.4 To determine virus production in surfection 3.5 To determine plasmid prepared method 3.6 To evaluate the shRNA effects on drug sensitivity in SAS. Chapter 4 Discussion 41 Chapter 5 Future Perspectives 48 Figures 50 References 74 | |
| dc.language.iso | en | |
| dc.subject | 魚藤素 | zh_TW |
| dc.subject | 口腔癌細胞 | zh_TW |
| dc.subject | 短髮夾核醣核酸 | zh_TW |
| dc.subject | deguelin | en |
| dc.subject | oral cancer cell | en |
| dc.subject | shRNA | en |
| dc.title | 在口腔癌細胞株中利用短髮夾核醣核酸進行綜合性致死篩選 | zh_TW |
| dc.title | Synthetic lethal and shRNA screen in oral cancer cell lines | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 洪志遠 | |
| dc.subject.keyword | 口腔癌細胞,短髮夾核醣核酸,魚藤素, | zh_TW |
| dc.subject.keyword | oral cancer cell,shRNA,deguelin, | en |
| dc.relation.page | 93 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2008-08-01 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 口腔生物科學研究所 | zh_TW |
| 顯示於系所單位： | 口腔生物科學研究所 | |
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