孫家棟Shun, Chia-Tung臺灣大學:法醫學研究所鄭翠芬Cheng, Tsui-FenTsui-FenCheng2010-04-302018-07-132010-04-302018-07-132009U0001-0702200915151000http://ntur.lib.ntu.edu.tw//handle/246246/178360"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？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.n 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文摘要……………………………………………………………………………………………..ii文摘要………………………………………………………………………………………….....iii錄;……………………………………………………………………………………………..…...iv目錄;………………………………………………………………………………………………..v目錄;……………………………………………………………………………………………….vi一章 …………………………………………………………………………………………….1一節 緒論; ………………………………………………………………………………….1二節 研究動機………………………………………………………… …………………...2三節 研究目的………………………………………………………… ………………….. 2四節 文獻回顧………………………………………………………………………………2二章 材料;與方法………………………………………………………………………………..9一節 檢體來;源………………………………………………………………………………9二節 實驗方法……………………………………………………………………………..10三節 儀器與設備…………………………………………………………………………..13三章 實驗結果…………………………………………………………………………………14四章 討論;………………………………………………………………………………………15五章 結論;與展望………………………………………………………………………22六;章 參;考文獻…………………………………………………………………………………24目錄;一 病例;.A.之組織病理;學H&E染色 .40X ,左下角插圖.100X………………………… 30二 病例; B之組織病理;學H&E 染色 40X ,左下角插圖.100X…………………………...30三 病例; C之組織病理;學H&E 染色 40X ,左下角插圖.100X………………………......31四 病例;A STR-15基因型 石蠟;包埋組織與血液………………………………… 32五 病例;A 第二次石蠟;包埋組織實驗結果 ………………………………………….. 33六; 病例; A 突變發生在vWA 基因座………………………………………………… 34七 病例;B STR-15基因型 石蠟;包埋組織與血液……………………………............ 35八 病例; B 突變發生在FGA基因座…………………………………………………... 36九 病例;C STR-15基因型 石蠟;包埋組織與血液………………………………….. 37十 病例;C 第二次石蠟;包埋組織實驗結果……………………………………......... 38十一 病例; C 突變發生在 D21S11 基因座…………………………………………… 39十二 體突變 (Somatic Mutation) 之 圖解說;表目錄一 本研究乳癌病人的年齡分佈………………………………………………………….. 40二 本研究病人乳癌組織型態的分佈…………………………………………………… 40三 本研究病人的乳癌期別分佈……………………………………………………………41四 本研究實驗組與對照組結果……………………………………………………………41五 本研究STR-15型實驗組與對照組比對結果……………………………………………41六 癌症組織類別與STR-15型基因座發生突變之關係……………………………………42七 乳癌病人的年齡與突變之關係…………………………………………………………42八 乳癌期別與突變關係…………………………………………………………………….43九 發生突變的乳癌案例的年齡﹑期別與突變基因座分佈……………………………….43十 病例.A.基因座分佈………………………………………………………………… 45十一 病例.B 基因座分佈……………………………………………………………….. 45十二 病例.C 基因座分佈……………………………………………………………….. 46十三 部分商品化STR檢測試劑盒之識別率…………………………………………… 46十四 FORDIS 資料庫比對病例A,B,C 結果................................................................. 47錄 人體實驗委員會 (IRB) 通過文號...................................................................... 48application/pdf1688036 bytesapplication/pdfen-US乳癌石蠟包埋DNA萃取基因突變STR-15型breast cancerparaffin-embedded DNA extractiongene mutation15 STR loci[SDGs]SDG3[SDGs]SDG16http://ntur.lib.ntu.edu.tw/bitstream/246246/178360/1/ntu-98-R94452003-1.pdf