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研究生:鄭翠芬
研究生(外文):Tsui-Fen Cheng
論文名稱:乳癌石蠟包埋組織於人身鑑別的可適用性
論文名稱(外文):The Availability of Paraffin-Embedded Cancer Tissue in Personal Identification: A Model of Breast Cancer
指導教授:孫家棟孫家棟引用關係
指導教授(外文):Shun Chia-Tung
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:法醫學研究所
學門:醫藥衛生學門
學類:其他醫藥衛生學類
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:48
中文關鍵詞:乳癌石蠟包埋DNA萃取基因突變STR-15型
外文關鍵詞:breast cancerparaffin-embedded DNA extractiongene mutation15 STR loci
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"鑑定"是法醫的主要工作,然而法醫學的鑑定不是局限在死因之鑑定,在司法領域中尚有許多和法醫有相關的鑑定工作,例如:親子鑑定、血緣鑑定也稱人身鑑別、物證鑑定、傷痕鑑定,在面對種種來源的證物、血斑、精斑、唾液斑、汗斑、皮膚、毛髮、指甲,甚至牙齒、骨骼等組織檢體,大部份都是仰賴DNA的鑑定技術。其中組織包埋在石蠟中理論上其相對DNA是呈穩定狀態被貯存的,而且從石蠟中去萃取組織DNA來當作鑑定已不是困難的事,不過,值得探討的是,假如使用的蠟塊包埋組織來源是癌症組織,是否有可能因有基因突變(癌症細胞增生)而造成與本來個體之正常組織的DNA不一樣了呢?而它能否可用來當成作鑑定的標準?
本文的研究利用在一年期間,2007年8月~2008年6月搜集了50個外科切除的乳癌病患之癌組織來供作研究中的實驗組,並同時研究病人身體的其他正常組織如靜脈血液當對照組。在石蠟組織的 DNA萃取,只有36個達到可分析的量,另外有14位病人的石蠟組織的DNA萃取的STR (short Tandem Repeats 短串聯重複序列)表現不完全,所以剔除在本分析之外,故可分析率是72% ( 36/50) ,血液的DNA萃取則全數成功。結果發現有92 ﹪(33/36)其癌組織STR 15 基因座與其血液是完全相匹配,而有3人的蠟塊包埋組織發現有突變,突變率是8.% ( 3/36), 3位突變均只發生在單一個基因座上( single locus ),在血緣鑑定中,以STR分型來評量,只發生單一基因座的突變並不足以用來排除二者非來自於同一個體,必須有二個或以上的位點不一樣 才能懷疑來自不同個體。 所以藉由本研究結果, 似乎至少證明大部份癌變組織中的DNA與未發生癌變的同一人身上其他正常組織的DNA是相同的,或許在沒有其他可比對的情況下石蠟包埋癌症組織仍可以提供作為DNA鑑定及人身鑑別的輔助來源或參考。
“Investigation” is the major work of Forensic Medicine. The forensic identification is not only restricted to realize the reason of death, but also involves a lot of judicial investigations, such as parentage analysis, paternity test or personal identification and biological evidence examination. Most of the forensic evidence examination, for example, blood, sperm, sweat, skin, hair, nail and even teeth and bone, relies on DNA analysis technology. Short tandem repeat (STR) system is one of the main methodologies used in criminal investigation activities. Evidences have shown that DNA in paraffin-embedded tissues is relatively stable, and it is not difficult to extract DNA from paraffin-embedded tissues. Furthermore, it has been proved that cancer is induced by accumulation of genetic mutations. Thus, it is worthy to consider that if the paraffin-embedded cancer tissues could provide a model of forensic identification.
In this study, we assayed 50 paraffin-embedded breast cancer tissues, which were collected from August 2007 to June 2008, and their counterpart blood specimens as control groups. The extraction rates of DNA were 72% (36/50) in paraffin and 100% in blood specimens, respectively. We revealed that 92 % (33/36) of 15 STR loci polymorphism in cancer tissues was perfectly matched with blood samples. The rest of 3 paraffin-embedded specimens DNA were found to be mutated from control group, which the mutation rate was approximately 8% (3/36). In addition, we also discovered that these 3 tissues have the same single locus mutation. Using STR system to evaluate the parentage relationship, single locus mutation is not enough to exclude the possibility that the two specimens are from the identical source. More than two loci of different are needed. Thus, these results demonstrate that DNA in most of cancer tissues is not mutated from normal parts, indicating that in some circumstances, paraffin-embedded cancer tissues might provide usefulness source of DNA analysis and forensic personal identification.
口試委員會審定書…………………………………………………………………………………...i
中文摘要……………………………………………………………………………………………..ii
英文摘要………………………………………………………………………………………….....iii
目錄……………………………………………………………………………………………..…...iv
圖目錄………………………………………………………………………………………………..v
表目錄……………………………………………………………………………………………….vi
第一章 …………………………………………………………………………………………….1
第一節 緒論 ………………………………………………………………………………….1
第二節 研究動機………………………………………………………… …………………...2
第三節 研究目的………………………………………………………… ………………….. 2
第四節 文獻回顧………………………………………………………………………………2
第二章 材料與方法………………………………………………………………………………..9
第一節 檢體來源………………………………………………………………………………9
第二節 實驗方法……………………………………………………………………………..10
第三節 儀器與設備…………………………………………………………………………..13
第三章 實驗結果…………………………………………………………………………………14
第四章 討論………………………………………………………………………………………15
第五章 結論與展望………………………………………………………………………22
第六章 參考文獻…………………………………………………………………………………24

圖目錄
圖一 病例.A.之組織病理學H&E染色 .40X ,左下角插圖.100X………………………… 30
圖二 病例 B之組織病理學H&E 染色 40X ,左下角插圖.100X…………………………...30
圖三 病例 C之組織病理學H&E 染色 40X ,左下角插圖.100X………………………......31
圖四 病例A STR-15基因型 石蠟包埋組織與血液………………………………… 32
圖五 病例A 第二次石蠟包埋組織實驗結果 ………………………………………….. 33
圖六 病例 A 突變發生在vWA 基因座………………………………………………… 34
圖七 病例B STR-15基因型 石蠟包埋組織與血液……………………………............ 35
圖八 病例 B 突變發生在FGA基因座…………………………………………………... 36
圖九 病例C STR-15基因型 石蠟包埋組織與血液………………………………….. 37
圖十 病例C 第二次石蠟包埋組織實驗結果……………………………………......... 38
圖十一 病例 C 突變發生在 D21S11 基因座…………………………………………… 39
圖十二 體突變 (Somatic Mutation) 之 圖解說表目錄

表一 本研究乳癌病人的年齡分佈………………………………………………………….. 40
表二 本研究病人乳癌組織型態的分佈…………………………………………………… 40
表三 本研究病人的乳癌期別分佈……………………………………………………………41
表四 本研究實驗組與對照組結果……………………………………………………………41
表五 本研究STR-15型實驗組與對照組比對結果……………………………………………41
表六 癌症組織類別與STR-15型基因座發生突變之關係……………………………………42
表七 乳癌病人的年齡與突變之關係…………………………………………………………42
表八 乳癌期別與突變關係…………………………………………………………………….43
表九 發生突變的乳癌案例的年齡、期別與突變基因座分佈……………………………….43
表十 病例.A.基因座分佈………………………………………………………………… 45
表十一 病例.B 基因座分佈……………………………………………………………….. 45
表十二 病例.C 基因座分佈……………………………………………………………….. 46
表十三 部分商品化STR檢測試劑盒之識別率…………………………………………… 46
表十四 FORDIS 資料庫比對病例A,B,C 結果................................................................. 47
附錄 人體實驗委員會 (IRB) 通過文號...................................................................... 48
1. Jeffreys AJ, Wilson V, Thein SL, Hypervariable ''minisatellite''regions in human DNA. Nature, 1985; 314:67-73
2. Jeffreys AJ, Royle NJ, Wilson V, Wong Z, Spontaneoumutation rates to new length alleles at tandem-repetitive hypervariable loci in human DNA. Nature, 1988; 332:278-281
3. Thyagarajan B., Anderson K E, Kong F , Selk F R, Lynch C F . New approaches for genotyping paraffin wax embedded breast tissue from patients with cancer: the Iowa women’s health study. J. Clin. Pathol. 2005; 58: 955-961.
4. Lipponen P, Saarelainen E, Ji H, Aaltomaa S, Syrjanen K. Expression of E-cadherin (E-CD) as related to other prognostic factors and survival in breast cancer. J Pathol. 1994; 174:101-109.
5. Serth J, Kuczyk MA, Paeslack U, Lichtinghagen R, JonasU. Quantitation of DNA extracted after micropreparationof cells from frozen and formalin-fixed tissue sections. Am J Pathol 2000; 156: 1189–1196.
6. Paik Soonmyung, Kim Chung-yeul, Song Yong-kuk and Kim. Wan-seop Technology Insight: application of molecular techniques to formalin-fixed paraffin-embedded tissues from breast cancer. Cooperative Human Tissue Network. Am J Clin Pathol 2002: 118: 733–741.
7. Isidre Ferrer, Judith Armstrong, Sabina Capellari, Piero Parchi, Thomas Arzberger, Jeanne Bell, Herbert Budka, Thomas Strobel, Giogio Giaccone, Giacomina Rossi, Nenad Bogdanovic, Peter Fakai, Andrea Schmitt, Peter Riederers ,Safa Al-Sarra, Rivka Ravid, Hans Kretzschmar. Effects of Formaline Fixation, Paraffin Embedding, and Time of Storage on DNA Preservation in Brain Tissue: A BrainNet Europe Study. Brain Pathol 2007; 17: 297-303
8. Farid Ahmed Abu-Bedair, Basiouny Ahmed El-Gamal, Nagi Ali Ibrahim and Abdelbaset Anwer El-Aaser. Serum Lipids and Tissue DNA Content in Egyptian Female Breast Cancer Patients . Jpn J Clin Oncol .2003; 33(6): 278-82
9. Bonin S, Petrera F, Rosai J, Stanta G. DNA and RNA obtained from Bruin’s fixed tissues. J Clin Pathol 2005; 58:313–316.
10. Gang Wang, Elizabeth Maher, Cameron Brennan, Lynda Chin, Christopher Leo, Manjit Kaur, Penny Zhu, DNA amplification method tolerant to sample degradation Genome Res. 2004; 14: 2357-2366
11. Simon A Joosse, Erik H van Beers, Petra M Nederlof.Automated array-CGH optimized for archival formalin-fixed, paraffin-embedded tumor material. BCM Cancer 2007; 7: 43
12. Lof M, Sandin S, Lagiou P, Hilakivi-Clark L, Trichopoulos D, Adami H-O, Weiderpass E. Dietary fat and breast cancer risk in Swedish women’s lifestyle and health cohort. Br J Cancer 2007;97:15701576
13. Karin B Michels .The contribution of the environment (especially diet) to breast cancer risk. Breast Cancer Res 2002; 4:58-61
14. Thierry Soussi .The p53 Mutation Handbook, Version 1.0 02/2007
15. Harris CC, Hollstein M. Clinical implications of the p53 Tumor-suppressor Gene. N Engl J Med. 1993; 329:1318-27
16. Chia-Tung Shun, Jaw-Town Lin, Shih-Pei Huang, Min-Tsan Lin, Ming-Shiang Wu,.
Expression of macrophage migration inhibitory factor is associated with enhanced angiogenesis and advanced stage in gastric carcinomas. World J Gastroenterol 2005; 11(24): 3767-3771.
17. Edwards A Civitello A, Hammond HA, Caskey CT DNA typing and genetic mapping with trimeric and tetrameric tandem repeats. Am J Hum Genet 1991; 49:746-756
18. 柯滄銘 ; 許家愷 ; 潘亨文 ; 徐碧梅 ; 鍾宜珮. 利用短縱列重複序列進行親子與手足關係的鑑定 中央警察大學學報,民國 90年第38期, pp 375-390
19. Andersen, J.F, Greengage M.J, Butler, H.R. Butle, Kilpatrick, S.R, Percy, R.C, Way, K.A, Mayhill, H.S, Wright, J.C, Hallett, R, Parkin, B.H. Further validation of a multiplex STR system for use in routine forensic identity testing. Forensic Sci. Int. 1996; 78:47–64
20. Jeffreys AJ, Wilson V, Thein I SL Individual-specific ''fingerprints'' of human DNA Nature 1985; 316:76-79
21. Urquhart A., Kimpton C. R, Downes T.J, Gill R, Variation in Short Tandem Repeat sequences: a survey of twelve microsatellite loci for use as forensic identification markers. Int J Legal Med 1994; 107: 13-20
22. Lygo JE, Johnson RE, Holdaway DJ, Woodroffe S, Whitaker JR, Clayton TM, Kimpton CR - R Gill, The validation of short tandem repeat (STR) loci for use in forensic casework. Int J Legal Med 1994; 107:77-89
23. W.Bar, Brinkmann B, Lincoln P, Mayr W.R, Rossi .U DNA Commission of the International Society for Forensic Haemogenetics, DNA recommendations -1994 report concerning further recommendations of the DNA Commission of the ISFH regarding PCR-based polymorphisms in STR (short tandem repeat) systems. Int J Legal Med. 1994; 107:159~160.
24. James Chun-I Lee, Chia-Hwei Chen, Li-Chin Tsai, and Jan-Gowth Chang .The Screen of 13 short tandems repeat loci in the Chinese population. Forensic Sci Int.1997; 87:137-144
25. James Chun-I Lee , Marie Lin, Li-Chin Tsai, Population study of polymorphic microsatellite DNA in Taiwan .Forensic Sci Journal. 2002; 1:31-37
26. Chih-Wei Wang, Ding-Ping Chen, Chi-Yuan Chen, Shu-Chuan Lu, Chien –Feng Sun. STR data for the AmpFISTR SGM Plus and Profiler loci from Taiwan. Forensic Sci Int. 2003; 138: 119-122
27. Nu En Hung, Ranajit Chakra borty, Bruce Budowle.DSS80 allele frequencies in a Chinese population. Int Leg Med 1994; 107:118-120
28. 法醫學 常林 主編 中國人民大學出版社 2008 第一版 第十七章 pp.239-247
29. 法醫物證案例疑難解析 斐黎 王海生 編著 中國人民公安大學出版社 2007 第一版 pp . 121-164
30. S. Eva Singletary, Craig Allred, Pandora Ashley, Lawrence W. Bassett, Donald Berry, Kirby I. Bland, Patrick I. Borgen, Gary Clark, Stephen B. Edge, Daniel F. Hayes, Lorie L. Hughes, Robert V.P. Hutter, Monica Morrow, David L. Page, Abram Recht, Richard L. Theriault, Ann Thor, Donald L. Weaver, H. Samuel Wieand, Frederick L. Greene Revision of the American Joint Committee on Cancer Staging System for Breast Cancer J Clin Oncol. 2002; 20: 3628-3636.
31. Rengul Cetin-Atalay , Mehmet Ozturk. p53 mutations as fingerprints of environmental carcinogens. Pure Appl. Chem. 2000; 72(6): 995–999.
32. Sofia Kouidou A, Andigoni Malousi A, Anastasios Kyventidis A, Aikaterini Fragou
A, Nicks Maglaveras G: C [ A: T mutations and potential epigenetic regulation of p53 in breast cancer. Breast Cancer Res Treat 2007; 106:351–360
33. Chappuis P, Goffin J, Wong N, Perret C, Ghadirian P, Tonin P, Foulkes W . A significant response to neoadjuvant chemotherapy in BRCA1/2 related breast cancer. J Med Genet. 2002; 39(8): 608–610
34 Gianni L, Zambetti M, Clark K, Baker J, Cronin M, Wu J, Mariani G, Rodriguez J, Carcangiu M, Watson D, Valagussa P, Rouzier R, Symmans WF, Ross JS, Hortobagyi GN, Pusztai L, Shak S Gene expression profiles in paraffin-embedded core biopsy tissue predict response to chemotherapy in women with locally advanced breast cancer. J Clin Oncol. 2005; 23(10): 7265-7277.
35. Mo11er A, Meyer E, Brinkmann B, Different types of structural variation in STRs HumFES/FPS, HumVWA and HumD21S11. Int J Legal Med. 1994; 106:319-323
36. 霍振义,刘雅诚,唐晖.2718例亲子关系鉴定结果的分析. 中國法醫學雜誌. 2003;18(1):31-32.
37 Kiyoshi Minaguchi, Osamu Takenaka.Structural variations of vWA locus in humans and comparison with non-human primates. Forensic Sci Int.2000; 113:9-16.
38. Matti Lukka, Gunnar TasA, Pekka Ellonen, Krisi Moilanen, Vitali Vassiljev, Ismo Ulmanen. Triallelic pattern in STR loci used for paternity analysis: Evidence for duplication in chromosome 2 containing the TPOX STR locus. Forensic Sci Int. 2006; 164:3-9.
39. Hristopher Vidal, Marisa Cassar. A Case of tri-allelic pattern at locus DS3S1358 on chromosome 3p21 inherited from paternal grandmother. Forensic Sci Int. Genetics Suppl Series 2008; 2:372-375
40. Tim M.Clayton, Jennifer L.Guest, Andy J.Urquhart, Peter D.Gill.A Genetic Basis for
Anomalous Band Patterns Encountered During DNA STR Profiling. J Forensic Sci.2004; 49 (6): 1207-1213.
41. Cecelia A. Crouse, Sue Rogers, Elizabeth Amiott, Sandra Gibson, Arni Masibay. Analysis and Interpretation of Short Tandem Repeat Microvariants and Three-Banded Allele Patterns Using Multiple Allele Detection Systems. J Forensic Sci.1998: 87-94.
42. Holly A. Hammond, Li Jin Y. Zhong, C. Thomas Caskey, Ranajit Chakraborty Evaluation of 13 Short Tandem Repeat Loci for Use in Personal Identification Applications. Am. J. Hum. Genet. 55:175-189, 1994
43 Dauber E.M, Dorner G, Wenda S, Schwartz-Jungl E.M, Glock B, Bar W, Mayr.W.A Unusual FGA and D19S433 off-ladder alleles and other allelic variants at the STR loci D8S1132, vWA, D18S51 and ACTBP2 (SE33). Forensic Sci Int. Genetics Suppl Series 2008; 1: 109-111.
44. Sabita K. Murthy, Lisa M. DiFrancesco, R. Travis Ogilvie, Douglas J. Demetrick, Loss of Heterozygosity Associated with Uniparental Disomy in Breast Carcinoma. Mod Pathol 2002; 15(12): 1241–1250
45. 實用法醫學司法鑑定 莫耀南 主編 北京 科學出版社 2008 第十五章 pp 339-401
46. DNA鑑定 ---PCR PCR in Forensic DNA Anlysis 李俊億 著 1995 第三章 pp.43-62; 第十一章 pp.173-182.
47. Banaschak S, Chesen A.Du, Brinkmann B. Multiple interchanging of tissue samples in case of breast cancer. Forensic Sci Int. 2000; 113: 3-7.
48. Dora A.Popiolek, Mechthild K.Prinz, A.Brain west, Bianca L.Nazzaruolo, Sheila M.Estacio, Zoran M.Budimlija. Multiplex DNA Short Tandem Repeat Analysis a useful Method for Determining the Provenance of Minute Fragments of Formalin-Fixed, Paraffin Embedded Tissue. Am J Clin Pathol 2003; 120:746-751.
49. C. M. Hsu, N. E. Huang, L. C. Tsai, L. G. Kao, C. H. Chao, A. Linacre ,J. C.-I. Lee. Identification of victims of the 1998 Taoyuan Airbus crash accident using DNA analysis. Int J Legal Med . 1999 ; 113 : 43–46
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