2011-05-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656708摘要：肺腺癌佔了非小細胞肺癌細胞型態中的最高比例，從 53-70% 不等。有別於白種人，肺腺癌在東亞人種有較高的表皮生長因子受體突變，以及對表皮生長因子受體抑制劑治療較高的反應率。肺腺癌的致病機轉至今不明。近來有研究提出多階段致病機轉的可能性：從異型腺瘤性增生到細支氣管肺泡腺癌，再進展到侵犯性肺腺癌。煙草的暴露現今被認為是誘發肺癌的危險因子，然而造成非吸煙族群肺腺癌的誘發因子，至今不明。現今許多癌症已知和感染症有關。最近已知羊的肺腺癌是由 jaagsiekte sheepretrovirus 引起並且會經由羊奶傳染給其它羊隻。近來不少病毒基因在人類肺腺癌檢體中被發現，並且被認為可能和肺癌的發生有關。這些包含了humanpapillomavirus 16/18, JC virus, jaagsiekte sheep retrovirus, 以及measles virus。然而，截至目前的研究都是針對某幾株已知病毒所進行的研究，並未有廣泛性微生物的偵測實驗。為瞭解肺癌的發生是否與微生物感染有關聯，我們利用現在所有已知的病毒及細菌基因體序列進行分析，建立病毒和細菌檢測探針的基因庫，並已成功發展微生物檢測晶片。該微生物晶片共有44000 探針，可偵測至少1000 種病毒及細菌。利用該微生物晶片技術，我們可以在一次的實驗中篩檢上千種的微生物。本計畫前兩年已完成晶片設計與客製，蒐集80 個肺腺癌檢體並完成核酸萃取，並建立微生物晶片以及高量高速定序的標準實驗流程與分析方法。利用多種癌症細胞株，所建立之標準實驗流程已被證實其實際可行性，並且在實際人類肺癌檢體中篩選出可能的致病相關病毒。經由前述實驗的成功執行，目前已完成本計畫前兩年之預期目標與結果。<br> Abstract: Adenocarcinoma accounts for the highest proportion of non-small cell lungcancer, ranging from 53-70%. It is a distinct disease entity in East Asian population inthat there is a significantly high mutation rate of epidermal growth factor receptor.Although cigarette smoking is believed to be an important risk factor in lung cancerdevelopment, the initiating factors of cell malignant transformation in non-smokerpulmonary adenocarcinoma are not yet found.Recent studies point out the possible inverse correlation between the hostimmune status and the risk of lung cancer development. Aside from clinical andepidemiological studies, researchers have also identified several microbes fromhuman pulmonary adenocarcinoma specimens, such as human papillomavirus 16/18,JC virus, jaagsiekte sheep retrovirus, adenovirus, and measles virus. All these cluessuggest the association between infections and pulmonary adenocarcinomadevelopment. However, previous studies were all designed to detect specific viruses,and no means have been used to perform generalized microbial identification in lungcancers.In order to perform a comprehensive investigation of microbial etiology in lungcancers, we have designed a set of microarray probes encompassing all viral andbacterial sequences archived in GenBank for microbial identification in humanpulmonary adenocarcinoma. The microbial array features more than 40,000 probesand is able to interrogate the presence of more than 1000 species of viruses andbacteria in the test samples. We have also collected 80 lung adenocarcinoma tissuesand have extracted the total RNA and genomic DNA from the tissues. Theexperimental protocols, from sample preparation, microbial microarray hybridization,RT-PCR and Sanger sequencing validation, to microbial microarray sequence capturefollowed by next-generation sequencing have been established. We have verified theprocedural pipeline of the experiment by using cancer cell lines known to harborviruses (XMRV in CWR22 and HPV18 in HeLa cell-lines) and then applied thewhole procedure to human non-smoker pulmonary adenocarcinoma tissues. Weobtained several putative oncogenic viruses in our first-stage experiments and needfurther in-depth verification. All in all, we have completed the milestones outlined inthe original proposal for the first two years.epithelial-mesenchymal cell interactionstooth regenerationbone morphogenic proteinsfibroblast growth factors