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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊雅倩 | |
| dc.contributor.author | Wei-Ting Weng | en |
| dc.contributor.author | 翁瑋廷 | zh_TW |
| dc.date.accessioned | 2021-06-13T01:15:38Z | - |
| dc.date.available | 2021-12-31 | |
| dc.date.copyright | 2011-10-07 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-08-03 | |
| dc.identifier.citation | 1. http://www.who.int/en/.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/29707 | - |
| dc.description.abstract | 大腸直腸癌目前不論在全世界或是台灣皆是癌症死亡原因的第三名。實驗室先前針對大腸直腸癌腫瘤檢體進行第四號染色體微衛星標記的失異合性(loss of heterozygosity)分析，發現於染色體4q25-4q28.3間，具有3個較高的基因刪除區域，本論文所探討之PCDH10基因即位於4q28.3。近年來的研究顯示，PCDH10在許多癌症中可能扮演抑癌基因的角色，而本實驗室則研究PCDH10於大腸直腸癌所扮演的角色。首先，PCDH10於大腸直腸癌腫瘤組織的mRNA表現量明顯低於其成對之正常大腸黏膜組織。而針對PCDH10進行體外功能的探討，發現當暫時性表現PCDH10於大腸直腸癌細胞株時，可以抑制癌細胞的生長、胞落形成以及侵犯能力，支持PCDH10對於大腸直腸癌可能為抑癌基因。本論文利用挑選穩定表現PCDH10之HCT116細胞，針對PCDH10進行一系列體外及動物活體內之功能性探討，研究結果顯示：當PCDH10再表現於HCT116細胞時，在體外的實驗觀察到PCDH10可以抑制癌細胞之生長、胞落形成、移動及侵犯能力；在動物活體內的實驗觀察到PCDH10可以抑制HCT116細胞於SCID小鼠皮下之腫瘤生成以及肝臟轉移能力，並進而使小鼠的存活率上升。為進一步探討PCDH10的抑癌機制，針對細胞凋亡、細胞週期及細胞老化做分析，在細胞凋亡的實驗結果顯示：PCDH10可以增加HCT116細胞之自發性細胞凋亡；在細胞週期的實驗結果顯示：當PCDH10再度表現會減緩HCT116細胞自G1進入S的細胞週期；在細胞老化的實驗結果顯示：PCDH10可以增HCT116細胞的老化作用。此外，PCDH10可透過調控p53表現量增加進而提升細胞週期抑制因子(cell cycle inhibitor) p21的蛋白表現量，導致HCT116細胞減緩自G1進入S的細胞週期及增加細胞老化。綜合以上結果可以進一步支持PCDH10在大腸直腸癌化過程中具有抑癌的功能。 | zh_TW |
| dc.description.abstract | Colorectal cancer (CRC) is the third leading cause of cancer death in the world and Taiwan. We had used loss of heterozygosity study to detect genetic deletion of chromosome 4 in CRC, and found out three regions with higher frequencies of genetic losses at 4q25-4q28.3. Protocadherin 10 (PCDH10) is located at 4q28.3, one of the three common deletion regions defined. In addition, PCDH10 mRNA expression in CRC tumors was significantly lower than the matched normal mucosas. Functional studies showed that transient expression of PCDH10 in HCT116 cells reduced cell proliferation, colony formation and invasion in vitro, suggesting that PCDH10 is as a tumor suppressor in CRC. In the present study, single stable clones of PCDH10 re-expression in HCT116 were selected. We used these cell clones to identify PCDH10 function via in vitro and in vivo assays. The in vitro studies showed that cell proliferation, colony formation, migration and invasion were significantly reduced by re-expression of PCDH10 in HCT116 cells. We also found that PCDH10 had the tumor suppressor function in tumorigenesis and liver metastasis in xenograft tumor models of SCID mice. To study tumor suppression mechanism mediated by PCDH10, apoptosis, cell cycle and senescence analysis were performed in the PCDH10-expressing single stable clones. Re-expression of PCDH10 increased spontaneous apoptosis of HCT116 cells. As for cell cycle analysis, PCDH10 re-expression delayed cell cycle from G1 phase to S phase in HCT116 cells after serum starvation. The senescence study indicates that PCDH10 expression can induce senescence in HCT116 cells. Futhermore, we found that PCDH10 can upregulate p53 protein, and then lead to p21-mediated retardation of G1 to S transition and senescence in HCT116 cells. In conclusion, the results from in vitro and in vivo assays support PCDH10 might play a tumor suppressor role in colorectal tumorigenesis and metastasis. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T01:15:38Z (GMT). No. of bitstreams: 1 ntu-100-R98424020-1.pdf: 3333097 bytes, checksum: 0aaab03e2a5b21a0824799badd3454fb (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | 中文摘要 i
英文摘要 ii 縮寫表 iv 1. 研究背景 1 1.1. 大腸直腸癌 1 1.2. 大腸直腸癌的分期 2 1.2.1. Dukes分期系統 2 1.2.2. TNM分期系統 3 1.2.3. AJCC /UICC分期系統 4 1.3. 大腸直腸癌的生成機制 4 1.3.1. 染色體的不穩定性 4 1.3.2. 微衛星不穩定性 5 1.3.3. CpG島甲基化表現型 5 1.4. 抑癌基因 5 1.5. 細胞計畫性死亡 6 1.5.1. 細胞凋亡 6 1.5.2. 細胞自噬 8 1.5.3 細胞壞死 8 1.6. 細胞週期 8 1.6.1 Cyclin、CDK以及細胞週期抑制因子 9 1.6.2. 細胞週期檢查點 9 1.7. 細胞老化 10 1.8. Protocadherin 10 (PCDH10) 10 1.8.1. Protocadherin家族 10 1.8.2. PCDH10之結構 11 1.8.3. PCDH10先前之研究 11 1.8.4. PCDH10與癌症相關之研究 11 1.9. 第4號染色體於癌症之研究 12 1.10. 實驗室先前之相關研究結果 13 2. 研究目標 14 3. 材料與方法 15 3.1. 細胞培養 15 3.2. 動物來源 15 3.3. 試劑、材料及抗體 15 3.4. 穩定表現株之篩選 16 3.5. 蛋白質的抽取及定量 17 3.6. 西方點墨法 17 3.7. 細胞增生分析 17 3.8. 胞落形成試驗 18 3.9. 細胞傷口癒合試驗 18 3.10. 基底膜基質侵犯能力試驗 18 3.11. 細胞凋亡分析 19 3.12. 細胞週期分析 19 3.13. 细胞老化分析 20 3.14. 異種移植腫瘤之小鼠模式 20 3.15. 脾臟內注射腫瘤細胞之小鼠模式 21 3.16. 病理組織處理及染色 21 3.16.1 病理組織處理 21 3.16.2. 蘇木紫-伊紅染色 21 3.16.3. 免疫組織化學染色 22 3.17. 統計分析 23 4. 結果 24 4.1. 體外分析(in vitro assay)：PCDH10 於大腸直腸癌細胞株HCT116之抑癌功能探討 24 4.1.1. PCDH10使HCT116細胞之增生能力下降 24 4.1.2. PCDH10抑制HCT116細胞之胞落形成能力 24 4.1.3. PCDH10抑制HCT116細胞之移動能力 24 4.1.4. PCDH10抑制HCT116細胞之侵犯能力 25 4.1.5. PCDH10增加HCT116細胞之自發性細胞凋亡 25 4.1.6. PCDH10減緩HCT116細胞自G1進入S之細胞週期 25 4.1.7. PCDH10藉由p53訊息傳遞路徑調控p21進而減緩細胞週期由G1期進入S期 26 4.1.8. PCDH10透過調控p21表現進而增加HCT116細胞之細胞老化 27 4.2. 動物活體內分析(in vivo assay)：PCDH10 於SCID小鼠模式之抑癌功能探討 27 4.2.1. PCDH10減緩HCT116細胞於SCID小鼠皮下之腫瘤生成 27 4.2.2. PCDH10抑制HCT116細胞於SCID小鼠之肝臟轉移並進而延長小鼠存活率 27 5. 討論 29 6. 圖 33 7. 參考資料 52 8. 附錄 61 | |
| dc.language.iso | zh-TW | |
| dc.subject | 細胞老化 | zh_TW |
| dc.subject | 大腸直腸癌 | zh_TW |
| dc.subject | PCDH10 | zh_TW |
| dc.subject | 抑癌基因 | zh_TW |
| dc.subject | 細胞凋亡 | zh_TW |
| dc.subject | 細胞週期 | zh_TW |
| dc.subject | Colorectal cancer | en |
| dc.subject | Senescence | en |
| dc.subject | Cell cycle | en |
| dc.subject | Apoptosis | en |
| dc.subject | Tumor suppressor gene | en |
| dc.subject | PCDH10 | en |
| dc.title | PCDH10於大腸直腸癌之抑癌功能探討 | zh_TW |
| dc.title | Study on the tumor suppressor function of PCDH10 in colorectal cancer | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林淑萍,俞松良,蔡明宏 | |
| dc.subject.keyword | 大腸直腸癌,PCDH10,抑癌基因,細胞凋亡,細胞週期,細胞老化, | zh_TW |
| dc.subject.keyword | Colorectal cancer,PCDH10,Tumor suppressor gene,Apoptosis,Cell cycle,Senescence, | en |
| dc.relation.page | 83 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2011-08-03 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 醫學檢驗暨生物技術學研究所 | zh_TW |
| 顯示於系所單位： | 醫學檢驗暨生物技術學系 | |
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