2011-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/657506摘要：子宮頸癌多導因人類乳突瘤病毒（Human papilloma virus，HPV）感染，女性一生中有60-80%的機會可能感染該病毒，目前診斷子宮頸癌最常採用陰道抹片與陰道鏡檢查，進一步可以DNA核酸分子標幟的方式，亦即利用PCR將人類乳突病毒的DNA 序列訊號複製放大，再以不同區段的探針(primer probe)進行核酸雜交，偵測出人類乳突瘤病毒感染，並定出病毒的基因型。人類乳突瘤病毒是一種小分子（直徑55 nm）Papovavirus科的DNA病毒，目前已知的人類乳突瘤病毒有一百多種基因亞型，其中有三十餘種會感染人類生殖器官的皮膚及黏膜，造成各種疾病。許多的研究已經顯示子宮頸癌前病變跟高危險性的HPV基因型有關。目前已經知道的高危險的基因型有16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, 73等；低危險群有6, 11, 26, 42, 43, 44, 53, 54, 55, 62, 66等。在1百多型人類乳突病毒中，第16、18型的人類乳突病毒具有高危險性，大約70%子宮頸癌病患與這2型病毒感染有關，其他約15-25%的病患則與第52、55、58型有關。近來原子力顯微術（atomic force microscopy，AFM）實驗有效地被運用在量測個別分子的力曲線繪製(force mapping)。AFM技術被認為在分子結構上的研究是相當具有前景，且最能夠表現出的單分子力學實驗。我們過去多年來使用原子力顯微術在奈米醫學方面已有相當經驗。在本計劃中，我們將從奈米操控與量測具有高致癌性人類乳突病毒單分子生物結構力學探討。我們已初步開始著手研究生物奈米操控與癌細胞奈米診斷、病毒奈米牛頓力診斷的力學特性與理論推導。本計劃為三年期計劃，第一年預期完成: (1).分離與存純化具有高致癌性人類乳突病毒單顆粒;(2).奈米操控具有高致癌性人類乳突病毒單分子結構。第二年計劃預期執行:奈米量測具有高致癌性人類乳突病毒單顆粒的生物力學探討。第三年計劃預期完成並建立生物奈米操控與癌細胞-HPV奈米診斷平台，同時建立人類乳突病毒奈米牛頓力診斷的力學特性與推導理論。<br> Abstract: Human papillomavirus (HPV)-induced cervical carcinogenesis is proposed to be multi-step in nature. The major steps in cervical carcinogenesis include persistent infection of the metaplastic cervical epithelium with one or more of the oncogenic HPV infection, clonal progression of the infected epithelium to cervical precancer, and further invasion.1 Although these fundamental steps are well established, several new genetic and immunologic studies have shed light on the factors that influence each of these transitions. Over 150 different HPV subtypes have been identified so far, with a subset of these being classified as high risk for oncogenesis. Persistent infection with oncogenic HPV is the main cause of cervical cancer. Polymerase-chain-reaction (PCR)-based assays show that HPV DNA exist in around 90.7- 96.6% patients with cervical cancer and in 13.4 -15.6% control women.2 About the detected HPV types in patients, in descending order of frequency, are types 16, 18, 45, 31, 33, 52, 58, and 35. Fifteen HPV types are classified as high-risk types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, and 82); 3 are classified as probable high-risk types (26, 53, and 66); and 12 are classified as low-risk types (6, 11, 40, 42, 43, 44, 54, 61, 70, 72, 81). The most frequently detected high risk HPV types (HPV 16, 51, 52, and 59) are similar in male of different sites, which is compatible with the female incidence. The conformational transition between oncogenic HPV viral particles by using a single molecular measurement, it is now possible to study the mechanical properties of helical polymers. Recently, atomic force microscopy (AFM) has been used to measure the binding forces of antibody-antigen, receptor-ligand, and those between complementary DNA strands. In this project, we are going to use AFM to determine the oncogenic HPV viral particle to the nano-molecular level in correlated with clinical significance. In the first year project, we will focus on: 1) the purification and identification of high risk HPVs; 2) the application of AFM in HPVs. In the second year project, we will try to understand the structure-function relationship of HPV molecules and to design nano-mechanical devices based on viral chemistry and energetics of stability. In the 3rd year project, we will try to illustrate the nano-molecular level of cancer-HPV interaction. Hopefully, the present project will shed new lights on the nano-molecular mechanism in further clinical & biological platform of HPVs in human malignancy.人類乳突瘤病毒（Human papilloma virusHPV）原子力顯微術(atomic force microscopy)奈米牛頓力(nano-scale Newton)作用力繪製(force mapping)相互作用力(interaction force)Human papilloma virusatomic force microscopyforce mappingnano-scale Newtoninteraction force