(3.227.249.155) 您好!臺灣時間:2021/05/07 06:09
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果

詳目顯示:::

我願授權國圖
: 
twitterline
研究生:祝振興
研究生(外文):Chen-Hsing
論文名稱:使用p53與Caspase-3蛋白作為大腸癌預後指標之研究
論文名稱(外文):A Study of Using p53 and Caspase-3 Proteins as Prognosis Indicators in Colorectal Cancer
指導教授:周芬碧周芬碧引用關係
指導教授(外文):Fen-Pi Chou
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:生化暨生物科技研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:52
相關次數:
  • 被引用被引用:0
  • 點閱點閱:82
  • 評分評分:系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔系統版面圖檔
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
局部性大腸直腸癌(CRC)病患主要治療方法為手術切除,但其中仍有高達百分之30至70的局部復發率。此篇論文主要研究局部晚期大腸直腸癌患者接受術前化學放射治療後,觀察其細胞凋亡情形,以評估是否可以p53及Caspase-3蛋白做為病患的預後指標。臨床上選擇II或III期的大腸直腸癌患者,在病患進行診斷切片及在接受術前化放療後手術切除腫瘤時收集檢體。將檢體的部分做病理的檢驗及病理分期。以免疫組織化學染色方法觀察p53與Caspase-3蛋白於檢體中表現情形,比較兩者之表現模式是否有相關性。結果顯示:檢體中p53的表現相異性頗大,在診斷檢體可觀察到無表現(20%)、少量表現(12%)及顯著的表現(68%),其中有表現p53之診斷檢體在細胞中之表現位置比例分別為細胞核(5%)、細胞質(15%)及兩者都有(80%);在接受術前化放療後之手術檢體可觀察到p53無表現(52%)、少量表現(24%)及顯著的表現(24%),其中有表現p53之手術檢體在細胞中之表現位置比例分別為細胞核(42%)、細胞質(16%)及兩者都有(42%)。所有檢體中Caspase-3之表現較p53為弱,區域亦較小,在診斷檢體可觀察到無表現(52%)、少量表現(8%)及顯著的表現(40%),其中有表現Caspase-3之診斷檢體在細胞中之表現位置比例分別為細胞核(5%)、細胞質(5%)及兩者都有(90%);在接受術前化放療後之手術檢體則無觀察到Caspase-3表現(100%)。以連續切片觀察,同時表現p53與Caspase-3的診斷檢體可觀察到兩者表現位置有重疊(80%)及相反(20%)的情形。觀察同一病例可發現多數患者(62%)接受術前化放療後,手術檢體較診斷檢體之p53的表現顯著減少,在Caspase-3的表現方面也有半數以上的患者(56%)表現顯著減少,有些患者(31%)可共同觀察到此二蛋白的表現減少。有部分病患(45%)在接受術前化放療後,癌期症狀可觀察到有減緩的現象,其中又有患者(36%)診斷檢體之p53有顯著表現,而在接受術前化放療後,手術檢體之p53表現明顯減少。接受術前化放療後,在患者手術檢體上無觀察到Caspase-3表現。在此研究可觀察到術前化放療對大腸直腸癌患者,有使其病理趨緩的情形,且其p53表現亦有明顯減少,Caspase-3蛋白的表現甚至觀察到為完全消失。p53與Caspase-3的表現對於術前接受化放療病患具有其特定性,推估可以觀察患者之癌期變化,配合比較其p53與Caspase-3蛋白在術前化放療前後之表現量,作為預後評估之指標。

Surgery is the main curative modality in patients with localized colorectal cancer (CRC), but there is still about 30 to 70 % probability that local relapses occurred. The aim of the present work was to observe cell apoptosis of those patients with localized colorectal cancer who took preoperative chemoradiotherapy and evaluated the possibility that use p53 and Caspase-3 proteins as prognosis indicators on patients. Patients with clinical stage II or III colorectal cancer were selected for sample collection at the time of diagnosis or surgery. The pathology of cancer cell was recorded. Immunohistochemistry was performed to observe the expression of p53 and Caspase-3 proteins in tissue sections. Compared the p53 protein with Caspase-3 protein and analyzed the connections between these two proteins. Our data showed that the expressions of p53 were various differences between the tissue sections. In diagnosis sections, it was observed no detectable p53 (20%), few expressed p53 (12%) and significant expressed p53 (68%). For those diagnosis sections expressed p53, some were detected expression in nucleus only (5%), some were detected expression in cytoplasm only (15%) and some were detected in both nucleus and cytoplasm (80%). In surgery sections, which are collected after preoperative chemoradiotherapy, there were observed no detectable p53 (52%), few expressed p53 (24%) and significant expressed p53 (24%). For those surgery sections expressed p53, some were detected expression in nucleus (42%), some were detected expression in cytoplasm (16%) and some were detected in both nucleus and cytoplasm (42%). Compared with p53, the expressions of Caspase-3 in all sections were weaker and detected in smaller area. In diagnosis sections, there were observed no detectable Caspase-3 (52%), few expressed Caspase-3 (8%) and significant expressed Caspase-3 (40%). For those diagnosis sections expressed Caspase-3, some were detected expression in nucleus (5%), some were detected expression in cytoplasm (5%) and some were detected in both nucleus and cytoplasm (90%). In surgery sections, there was observed no detectable Caspase-3 at all (100%). Serial sections were showed that p53 and Caspase-3 proteins expressed at the same location in diagnosis sections (80%) and some of them expressed p53 and Caspase-3 proteins at the different location instead (20%). Compared diagnosis section with surgery section both of them were from the same patient, lots of the patients (62%) were observed that surgery sections had less expression of p53 than that in diagnosis sections and more than half of patients (56%) were observed that surgery sections had less expression of Caspase-3 than that in diagnosis sections. Some of those patients (31%) were observed that surgery sections had less expression both of p53 and Caspase-3 than that in diagnosis sections. It was observed that the pathology of colorectal cancer on patients (45%) who took preoperative chemoradiotherapy were downstaged. Some of them (36%) were detected significant expression of p53 in diagnosis sections and the expression of p53 was reduced significantly after preoperative chemoradiotherapy. There was no detectable Caspase-3 in surgery sections from patients after preoperative chemoradiotherapy. At the present work, we observed that the pathology on patients with colorectal cancer was downstaged, the expression of p53 was reduced significantly and the expression of Caspase-3 was disappeared after preoperative chemoradiotherapy. Expressions of p53 and Caspase-3 are specific to the patients who were treated with preoperative chemoradiotherapy. It was suggested that p53 and Caspase-3 can be the prognosis indicators by comparing the expressions of these two proteins with the stage of colorectal cancer before and after preoperative chemoradiotherapy.

中文摘要 2
Abstract 4
縮寫檢索表 6
壹、緒論 7
貳、實驗動機 15
參、材料與方法 16
一、 材料 16
(一) 檢體來源 16
(二) 免疫組織化學染色 16
二、 實驗方法 19
(一) 免疫組織化學染色 19
肆、結果 22
伍、討論 25
陸、圖表與圖表說明 28
柒、參考文獻 42
捌、附錄 49

Attallah, A.M., Abdel-Aziz, M.M., El-Sayed, A.M., Tabll, A.A. (2003) Detection of serum p53 protein in patients with different gastrointestinal cancers. Cancer Detect Prev 27, 127–131.
Baker, S.J., Fearon, E.R., Nigro, J.M., Hamilton, S.R., Preisinger, A.C., Jessup, J.M., van Tuinen, P., Ledbetter, D.H., Barker, D.F., Nakamura, Y., White, R., Vogelstein, B. (1989) Chromosome 17 deletions and p53 gene mutations in colorectal carcinomas. Science 244, 217-221.
Bertolini, F., Bengala, C., Losi, L., Pagano, M., Iachetta, F., Dealis, C., Jovic, G., et al. (2007) Prognostic and predictive value of baseline and posttreatment molecular marker expression in locally advanced rectal cancer treated with neoadjuvant chemoradiotherapy. Int J Radiat Oncol Biol Phys 68, 1455-1461.
Bertorelle, R., Esposito, G., Belluco, C., Bonaldi, L., Del Mistro, A., Nitti, D., Lise, M., Chieco-Bianchi, L. (1996) p53 gene alterations and protein accumulation in colorectal cancer. J Clin Pathol: Mol Pathol 49, M85-M90.
Bleday, R., Wong, W.D. (1993) Recent advances in surgery for colon and rectal cancer. Curr Probl Cancer 17, 1-68.
Broll, R., Duchrow, M., Oevermann, E., Wellm, C., Schwandner, O., Schimmelpenning, H., et al. (2001) p53 autoantibodies in sera of patients with a colorectal cancer and their association to p53 protein concentration and p53 immunohistochemistry in tumor tissue. Int J Colorectal Dis 16, 22–27.
Bruso, C.E., Shewach, D.S., Lawrence, T.S. (1990) Fluorodeoxyuridine-induced radiosensitization and inhibition of DNA double strand break repair in human colon cancer cells. Int J Radiat Oncol Biol Phys 19, 1411-1417.
Budihardjo, I., Oliver, H., Lutter, M., et al. (1999) Biochemical pathways of caspase activation during apoptosis. Ann Rev Cell Dev Biol 15, 269–290.
de Heer, P., de Bruin, E.C., Klein-Kranenbarg, E., Aalbers, R.I.J.M., Marijnen, C.A.M., Putter, H., de Bont, H.J., Nagelkerke, J.F., van Krieken, J.H.J.M., Verspaget, H.W., van de Velde, C.J.H., Kuppen, P.J.K. (2007) Caspase-3 activity predicts local recurrence in rectal cancer. Clin Cancer Res 13, 5810-5815.
Elsaleh, H., Powell, B., McCaul, K., Grieu, F., Grant, R., Joseph, D., et al. (2001) p53 alteration and microsatellite instability have predictive value for survival benefit from chemotherapy in stage III colorectal carcinoma. Clin Cancer Res 7, 1343–1349.
Forslund, A., Kressner, U., Lindmark, G., Inganas, M., Lundholm, K. (2001) Serum anti-p53 in relation to mutations across the entire translated p53 gene in colorectal carcinomas. Int J Oncol 19, 501–506.
Gatta, G., Faivre, J., Capocaccia, R., Ponz, de Leon M. (1998) Survival of colorectal cancer patients in Europe during the period 1978–1989. Eur J Cancer 34, 2176–2183.
Giralt, J., de las Heras, M., Cerezo, L., et al. (2005) The expression of epidermal growth factor receptor results in a worse prognosis for patients with rectal cancer treated with preoperative radiotherapy: Amulticenter, retrospective analysis. Radiother Oncol 74, 101-108.
Heidelberger, C., Griesbach, L., Montag, B.J., et al. (1958) Studies on fluorinated pyrimidines, II effects on transplanted tumors. Cancer Res 18, 305-317.
Huang, L.C., Clarkin, K.C., Wahl, G.M. (1996) Sensitivity and selectivity of the DNA damage sensor responsible for activating p53-dependent G1 arrest. Proc Natl Acad Sci 93, 4827-4832.
Jass, J.R., Ajioka, Y., Allen, J.P., Chan, Y.F., Cohen, R.J., Nixon, J.M., Radojkovic, M., Restall, A.P., Stables, S.R., Zwi, L.J. (1996) Assessment of invasive growth pattern and lymphocytic infiltration in colorectal cancer. Histopathology 28, 543-548.
Jass, J.R., Constable, L., Sutherland, R., Winterford, C.M., Walsh, M.D., Young, J., Leggett, B.A. (2000) Adenocarcinoma of colon differentiating as dome epithelium of gut-associated lymphoid tissue. Histopathology 36, 116–120.
Kastan, M.B. (1996) Signaling to p53: where does it all start? Bio Essays 18, 617-619.
Klionsky, D.J., Emr, S.D. (2000) Autophagy as a regulated pathway of cellular degradation. Science 290, 1717-1721.
Kobayashi, T., Masumoto, J., Tada, T., Nomiyama, T., Hongo, K., Nakayama, J. (2007) Prognostic significance of the immunohistochemical staining of cleaved caspase-3, an activated form of caspase-3, in gliomas. Clin Cancer Res 13, 3868-3874.
Krook, J.E., Moertel, C.G., Gunderson, L.L., et al. (1991) Effective surgical adjuvant therapy for high-risk rectal carcinoma. N Engl J Med 324, 709–715.
Lechpammer, M., Lukac, J., Lechpammer, S., Kovacevic, D., Loda, M., Kusic, Z. (2004) Humoral immune response to p53 correlates with clinical course in colorectal cancer patients during adjuvant chemotherapy. Int J Colorectal Dis 19, 114–120.
Li, S., Nie, Z., Li, N., Li, J., Zhang, P., Yang, Z., et al. (2003) Colorectal cancer screening for the natural population of Beijing with sequential fecal occult blood test: A multicenter study. Chin Med J 116, 200–202.
Luo, J.C., Neugut, A.I., Garbowski, G., Forde, K.A., Treat, M., Smith, S., et al. (1995) Levels of p53 antigen in the plasma of patients with adenomas and carcinomas of the colon. Cancer Lett 91, 235–240.
Matkowskyj, K.A., Cox, R., Jensen, R.T., and Benya, R.V. (2003) Quantitative immunohistochemistry by measuring cumulative signal strength accurately measures receptor number. J Histochem Cytochem 51, 205–214.
Miller, E.M. and Kinsella, T.J. (1992) Radiosensitization by fluorodeoxyuridine: Effects of thymidylate synthase inhibition and cell synchronization. Cancer Res 52, 1687-1694.
Minsky, B.D., Cohen, A.M., Kemeny, N., et al. (1992) Combined modality therapy of rectal cancer: Decreased acute toxicity with the preoperative approach. J Clin Oncol 10, 1218–1224.
Mrozek, A., Petrowsky, H., Sturm, I., Kraus, J., Hermann, S., Lorenz, M., Dorken, B., Daniel, P.T. (2003) Combined p53/Bax mutation results in extremely poor prognosis in gastric carcinoma with low microsatellite instability. Cell Death Differ 10, 461-467.
Nelson, W.G., Kastan, M.B. (1994) DNA strand breaks: the DNA template alterations that trigger p53-dependent DNA damage response pathways. Mol Cell Biol 14, 1815-1823.
NIH Consensus Conference. (1990) Adjuvant therapy for patients with colon and rectal cancer. JAMA 264, 1444–1450.
Rau, B., Sturm, I., Lage, S., et al. (2003) Dynamic expression profile of p21 WAF1/CIP1 and Ki-67 predicts survival in rectal carcinoma treated with preoperative radiochemotherapy. J Clin Oncol 21, 3391-3401.
Repetto, L., Venturino, A., Fratino, L., Serraino, D., Troisi, G., Gianni, W., et al. (2003) Geriatric oncology: A clinical approach to the older patient with cancer. Eur J Cancer 39, 870–880.
Rodrigues, N.R., Rowan, A., Smith, M.E.F., Kerr, I.B., Bodmer, W.F., Gannon, J.V. and Lane, D.P. (1990) p53 mutations in colorectal cancer. Proc Natl Acad Sci 87, 7555-7559.
Rudy, D.R., Zdon, M.J. (2000) Update on colorectal cancer. Am Fam Physician 61, 1759-1770, 1773-1774.
Russo, A., Migliavacca, M., Zanna, I., Valerio, M.R., Latteri, M.A., Grassi,N., Pantuso, G., Salerno, S., Dardanoni, G., Albanese, I., La Farina, M., Tomasino, R.M., Gebbia, N., Bazan, V. (2002) p53 mutations in L3-loop zinc-binding domain, DNA-ploidy, and S Phase fraction are independent prognostic indicators in colorectal cance: A prospective study with a five-year follow-up. Cancer Epidemiol Biomarkers Prev 11, 1322-1331.
Sandler, B., Smirnoff, P., Shani, A., Idelevich, E., Kaganski, N., Pfefferman, R., et al. (1999) The role of blood vessels of soluble 53 kDa protein and CEA in monitoring colon cancer patients. Anticancer Res 19, 4229–4233.
Schelwies, K., Sturm, I., Grabowski, P., Scherubl, H., Schindler, I., Hermann, S., Stein, H., Buhr, H.J., Riecken, E.O., Zeitz, M., Dorken, B., Daniel, P.T. (2002) Analysis of p53/BAX in primary colorectal carcinoma: Low BAX is a negative prognostic factor in UICC stage III tumors. Int J Cancer 99, 589-596.
Seifert, P., Baker, L.H., Reed, M.L., et al. (1975) Comparison of continuously infused 5-fluorouracil with bolus injection in treatment of patients with colorectal adenocarcinoma. Cancer 36, 123-128.
Shepherd, N.A., Hall, P.A., Coates, P.J., Levison, D.A. (1987) Primary malignant lymphoma of the colon and rectum. A histopathological and immunohistochemical analysis of 45 cases with clinicopathological correlations. Histopathology 12, 235-252.
Shepherd, N.A., Saraga, E-P., Love, S.B., Jass, J.R. (1989) Prognostic factors in colonic cancer. Histopathology 14, 613–620.
Shingleton, W.W., Prosnitz, L.R. (1985) Adjuvant therapy of colorectal cancer. Curr Probl Cancer 9, 1-34.
Shim, K.S., Kim, K.H., Park, P.W., Lee, S.Y., Choi, J.H., Han, W.S., et al. (1998) Increased serum levels of mutant p53 proteins in patients with colorectal cancer. J Korean Med Sci 13, 44–48.
Shoshana, P., Joachim, Y. (2006) Pathways that regulate autophagy and their role in mediating tumor response to treatment. Autophagy 2, 291-293.
Sturm, I., Kohne, C.H., Wolff, G., Petrowsky, H., Hillebrand, T., Hauptmann, S., Lorenz, M., Dorken, B., Daniel, P.T. (1999) Analysis of the p53/BAX pathway in the colorectal cancer: low BAX is a negative prognostic factor in patients with resected liver metastasis. J Clin Oncol 17, 1364-1374.
Sturm, I., Petrowsky, H., Volz, R., Lorenz, M., Radetzki, S., Hillebrand, T.,Wolff, G., Hauptmann, S., Dorken, B., Daniel, P.T. (2001) Analysis of p53/BAX/P16(ink4a/CDKN2)in esophageal squamous cell carcinoma: High BAX and p16(ink4a/CDKN2)identifies patients with good prognosis. J Clin Oncol 19, 2272-2281.
Takeda, A., Shimada, H., Nakajima, K., Yoshimura, S., Suzuki, T., Asano, T., et al. (2001) Serum p53 antibody as a useful marker for monitoring of treatment of superficial colorectal adenocarcinoma after endoscopic resection. Int J Clin Oncol 6, 45–49.
Tang, R., Ko, M.C., Wang, J.Y., Changchien, C.R., Chen, H.H., Chen, J.S., et al. (2001) Humoral response to p53 in human colorectal tumors: a prospective study of 1209 patients. Int J Cancer 94, 859–863.
Thiis-Evensen, E., Hoff, G.S., Sauar, J., Majak, B.M., Vatn, M.H. (1999) Flexible sigmoidoscopy or colonoscopy as a screening modality for colorectal adenomas in older age groups? Findings in a cohort of the normal population aged 63–72 years. Gut 45, 834–839.
Tulchinsky, H., Rabau, M., Shacham-Shemueli, E., Goldman, G., Geva, R., Inbar, M., Klausner, J.M., Figer, A. (2006) Can rectal cancers with pathologic T0 after neoadjuvant chemoradiation (ypT0) be treated by transanal excision alone? Annals of Surgical Oncology 13, 347-352.
van de Velde, C. J. H. (2005) Treatment of liver metastases of colorectal cancer. Annals of Oncology 16, ii144–ii149.
Vogelstein, B., Kinzler, K.W. (1993) The multistep nature of cancer. Trends in Genetics 9, 138-141.
Wang, Y., Singh, R., Massey, A.C., Kane, S.S., Kaushik, S., Grant, T., Xiang, Y., Cuervo, A.M., Czaja, M.J. (2008) Loss of macroautophagy promotes or prevents fibroblast apoptosis depending on the death stimulus. J Biol Chem 283, 4766-4777.
Yatabe, Y., Nakamura, S., Nakamura, T., Seto, M., Qgura, M., Kimura, M., Kuhara, H., Kobayashi, T., Taniwaki, M., Morishima, Y., Koshikawa, T., Suchi, T. (1998) Multiple polypoid lesions of primary mucosa-associated lymphoid-tissue lymphoma of colon. Histopathology 32, 116–125.
Zhang, Y.L., Zhang, Z.S., Wu, B.P., Zhou, D.Y. (2002) Early diagnosis for colorectal cancer in China. World J Gastroenterol 8, 21–25.


QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關論文
 
系統版面圖檔 系統版面圖檔