Prognostic significance of nm23-H1 expression in oral squamous cell carcinoma

Recent studies indicated nm23-H1 played a role in cancer progression. Therefore, we investigated clinical significance of nm23-H1 expression in oral squamous cell carcinoma (OSCC). In total, 86 OSCC specimens were immunohistochemically stained with nm23-H1-specific monoclonal antibodies. Immunohistochemical staining of nm23-H1 was confirmed by immunoblotting. The relations between nm23-H1 expression and clinicopathologic variables were evaluated by chi(2) analysis. As increased size of primary tumour could escalate metastatic potential and the data of patients at the late T stage might confound statistical analyses, we thus paid special attention to 54 patients at the early T stage of OSCC. Statistical difference of survival was compared by a log-rank test. Immunohistochemically, nm23-H1 expression was detected in 48.8% (42 out of 86) of tumorous specimens. It positively correlated with larger primary tumour size (P = 0.03) and inversely with cigarette-smoking habit (P = 0.042). In patients at the early T stage, decreased nm23 expression was associated with increased incidence of lymph node metastasis (P = 0.004) and indicated poor survival (P = 0.014). Tumour nm23-H1 expression is a prognostic factor for predicting better survival in OSCC patients at the early T stage, which may reflect antimetastatic potential of nm23. Therefore, modulation of nm23-H1 expression in cancer cells can provide a novel possibility of improving therapeutic strategy at this stage. In addition, our results further indicated cigarette smoking could aggravate the extent of nm23-H1 expression and possibly disease progression of OSCC patients. (C) 2004 Cancer Research UK

Detection of severe acute respiratory syndrome-associated coronavirus in pneumocytes of the lung

Previous reports have indicated that patients with severe acute respiratory syndrome (SARS)-associated coronavirus infection could develop atypical pneumonia with fulminant pulmonary edema. However, the target cells of SARS viral infection have not been characterized in detail. We report the pathologic findings of the lung in 3 cases of SARS. Chest radiographs at 2 to 3 weeks of infection revealed an atypical pneumonia with pulmonary consolidation, a clinical characteristic of SARS infection. The presence of the SARS virus was determined by nested reverse transcription polymerase chain reaction (RT-PCR), and the infected cells were identified by in situ hybridization in open-lung biopsy and postmortem necropsy specimens. Expression of SARS virus-encoded RNA was detected in all 3 cases by RT-PCR, and the SARS viral signal was localized in pneumocytes by using in situ hybridization

Correlation of Drug- and Radio-Resistance with Down-Regulation of Gene Expression of DNA Repair-Associated Proteins in Lung Adenocarcinoma Cells (III)

肺腺癌細胞之抗藥性及放射線耐受能力與DNA 修護作用相關基因異常表現之探討由上一個國科會研究計畫 (NSC92-2320-B-005-011) 中，我們發現 (1) 二氫二醇去氫.之表現與細胞對抗藥性與細胞耐放射線能力有關；(2) 肺腺癌細胞之抗藥性與細胞對放射線耐受能力可能與 DNA 修護作用有關；此外，(3) 當我們檢測肺腺癌細胞之基質金屬蛋白.群的差異表現時，更發現某些基質金屬蛋白.與二氫二醇去氫.之表現成正比。這種發現證實我們以前使用鑑別顯示法檢測非小細胞肺癌之新基因表現時，發現二氫二醇去氫.基因之高度表現，與其顯著預後價值有關 (Cancer Res 2001;61:2727-31)；此類關聯在第一期肺癌患者中也相當顯著 (Oncol Report 2002;9:515-9)。二氫二醇去氫.在癌細胞中的功能主要可能是去除抗癌藥之活性 (J Biol Chem 2002;277: 15035-43.)，此外，本類型之酵素能將前列腺素 D2 轉化為 9α,11β-前列腺素 F2，9α,11β-前列腺素F2 除了活性比前列腺素D2強許多倍之外，它又和新血管生成以及癌細胞之轉移能力有關。因此，本研究計畫將繼續研究肺腺癌細胞中二氫二醇去氫.之表現與細胞抗藥性、基質金屬蛋白.與腫瘤轉移能力之關係；在金計畫中更藥將本研究計畫的範圍擴充延伸到1. 探討肺腺癌細胞之抗藥性及放射線耐受力是否與 apoptosis inducing factor (AIF), checkpoint 2 protein (chk2) 和 Nejmegen break syndrome protein 1 (NBS1) 之基因表現有關；2. 檢測肺腺癌細胞中二氫二醇去氫.之表現，如何反映到 PGF 之濃度差異，並如何影響 AIF, chk2 和 NSB1 以及細胞抗藥性、放射線耐受力與腫瘤轉移之能力；3. 探討肺腺癌細胞對抗藥性及放射線耐受力如何與粒線體功能有關之基因如 AIF, NM23-H1 (nucleoside diposphate kinase, NDK), mitofusin 1/2 等交叉反應；4. 完成 AIF 之表現與細胞對抗藥性及放射線耐受力的研究，此研究重點將擺在 AIF之基因表現之調控，即以人為方法增加或降低肺腺癌細胞中AIF 基因之表現，並觀察其對肺腺癌細胞之抗藥性與細胞耐放射線能力之影響；5. 檢測所篩檢之抑制劑或刺激劑對肺腺癌細胞表現二氫二醇去氫.與 AIF, nm23 及 Mfn 之表現與抗藥性能力的關係；6. 探討所篩檢之抑制劑或刺激劑對肺腺癌細胞表現二氫二醇去氫.與 AIF, nm23 及 Mfn 之表現與細胞耐放射線能力的關係；和7. 探討肺腺癌細胞表現二氫二醇去氫.與前列腺素 F受體 peroxisome proliferator- activated receptor 及epidermal growth factor receptor (EGFR) 之表現與細胞中 Erk 之磷酸化程度的關係

Studies on Canine Oral/Nasal Tumors and EBV-Related Virus

EB病毒(Epstein-Barr virus, EBV)普遍感染人類，與人類多種腫瘤，如鼻咽癌(nasopharyngeal carcinoma, NPC)及某些口腔腫瘤(oral tumors)有關聯。EBV在分類上屬於丙型疱疹病毒(gammaherpesvirus, γ-HV)亞科之隱淋巴球病毒(Lymphocryptovirus, LCV)屬，是已知感染人類之唯一的LCV屬別病毒。我們先前研究證明犬普遍曾受到與EBV極相近病毒感染，許多犬具有特異抗體辨識EBV的蛋白，在一些犬隻白血球發現EBV病毒特有之DNA序列，以及發現有些骨骨遀細胞與白血球強烈表現與EBV潛伏感染有關的EBV特異RNA (EBV-encoded RNA, EBER)。我們的研究為首篇犬感染γ-HV之研究報導，且為首度發現LCV屬病毒有可能感染非靈長類動物。更值得注意的是，此研究開啟了將來後續探討EBV是否會在人與犬間傳播之重要研究議題，以及利用犬做為EBV感染之動物模式之可能性。最近我們繼續觀測陽性犬，發現其中一隻陽性犬於軟顎中發展出口腔腫瘤，我們此項的發現，首度揭示了犬口腔腫瘤與EBV相關病毒的可能關聯性。因此，本計畫擬進行犬口腔及鼻腫瘤(oral and nasal tumors)與EBV相關病毒之研究，調查犬常見的口腔腫瘤及犬鼻腫瘤帶有EBV相關病毒訊號(DNA序列、RNA表現)之情形，統計分析犬常見的口腔及鼻腫瘤與EBV相關病毒之關聯性，並致力於選殖EBV相關病毒基因或完整基因體，以及建立腫瘤細胞株。Epstein-Barr virus (EBV) is a widespread virus in human population. It is associated with several human malignancies such as nasopharyngeal carcinoma (NPC) or certain oral tumors. As a member of the lymphocryptovirus (LCV) genus of gammaherpesvirus (γ-HV), EBV is the only LCV known to infect human. Previously, we demonstrated that EBV-like virus infection could be frequently detected in the peripheral blood of pet dogs. We discovered that many dogs possessed antibodies against EBV-specific proteins. In several leukocyte DNA samples, we detected EBV-specific DNA sequence. Furthermore, EBV-encoded RNA (EBER) signal, which is often expressed during EBV latency, was detected in dog lymphocytes and bone marrow sections. This was the first report about canine γ-HV infection. Our data suggest that the host of LCV might not be restricted to the primates. Most of all, our study may reveal the possibility of zoonotic transmission or shed a light on the correlation of canine EBV-like virus infection with human disorders.Recently, oral tumor was developed in one dog that had been determined as positive for EBV-related DNA sequence. This observation further uncovers a possible link between canine oral tumors and EBV-related virus. In this project, we intend to explore EBV-related virus on canine oral and nasal tumors. We will investigate EBV-related viral signals (DNA sequence and RNA expression) on canine oral and nasal tumors. Association between any canine oral and nasal tumors will be examined by statistical analysis. In addition, we will proceed with molecular cloning of EBV-related viral sequences or complete genome and establishing tumor cell line

Studies on the Association between Canine EBV-Related Virus and Tumors

EB病毒(Epstein-Barr virus, EBV)普遍感染人類，與人類多種腫瘤例如淋巴瘤、鼻咽癌及某些口腔腫瘤有關聯。EBV在分類上屬於丙型疱疹病毒(gamma herpesvirus, γ-HV)亞科之隱淋巴球病毒(Lymphocryptovirus, LCV)屬，是已知感染人類之唯一的LCV屬別病毒。我們先前發表犬普遍曾受到與EBV極相近病毒感染，許多犬具有特異抗體辨識EBV的蛋白，在一些犬隻白血球DNA發現EBV病毒特有之BamH I W序列，並發現有些骨骨遀 及白血球表現與EBV潛伏感染有關的EBV特異RNA (EBV-encoded RNA, EBER)。我們的研究為首篇犬感染γ-HV之報導，且為首度發現LCV屬病毒有可能感染非靈長類動物。更值得注意的是，此研究開啟了將來後續探討EBV是否會在人與犬間傳播之重要研究議題，以及利用犬做為EBV感染之動物模式之可能性。最近我們在犬隻白血球又發現EBV致癌基因潛伏膜蛋白1 (latent membrane protein 1, LMP1) 之DNA序列，與BamHI H右讀碼1 (BamHI H rightward reading frame 1, BHRF1) (與bcl-2 同源)之基因表現。繼續觀測白血球攜帶EBV病毒特有訊號之陽性犬，發現其中一隻於軟顎中發展出口腔腫瘤，初步再篩檢13隻口腔或鼻腫瘤犬之檢體，其中高達8隻（61%）於檢體偵測到EBV之DNA序列，這些數據意指犬EBV相關病毒可能亦與犬腫瘤有關。為瞭解犬EBV相關病毒與腫瘤之關聯，本研究擬收集犬口腔腫瘤、鼻腫瘤、及淋巴瘤檢體，調查各腫瘤帶有犬EBV相關病毒訊號(DNA序列、RNA表現)之情形，致力於選殖EBV相關病毒基因以及培養腫瘤細胞，並統計分析犬EBV相關病毒與各腫瘤之關聯性。Epstein-Barr virus (EBV) is a widespread virus in human population. It is associated with several human malignancies such as lymphoma, nasopharyngeal carcinoma (NPC) or certain oral tumors. As a member of the Lymphocryptovirus (LCV) genus of gamma herpesvirus (γ-HV), EBV is the only LCV known to infect human. Previously, we demonstrated that EBV-related virus infection could be frequently detected in the peripheral blood of pet dogs. We discovered that many dogs possessed antibodies against EBV-specific proteins. In several leukocyte DNA samples, we detected EBV-specific BamH I W sequence. Furthermore, EBV-encoded RNA (EBER) signal, which is often expressed during EBV latency, was detected in dog lymphocytes and bone marrow sections. This was the first report about canine γ-HV infection. Our data suggest that the host of LCV might not be restricted to the primates. Most of all, our study may reveal the possibility of zoonotic transmission or shed a light on the correlation of canine EBV-related virus infection with human disorders.Recently, we identified the presence of latent membrane protein 1 (LMP1) DNA sequence and the gene expression of BamHI H rightward reading frame 1 (BHRF1) in canine leukocytes. Both LMP1 and BHRF1 are EBV-encoded oncogenes. Moreover, oral tumor was developed in one dog that had been determined as positive for EBV-related DNA sequence. We carried out a pilot study on 13 canine specimens of oral or nasal tumors and further identified 8 (61%) with EBV-specific DNA sequence. These observations further uncover a possible link between canine EBV-related virus and tumor development. In this project, we intend to explore whether canine EBV-related virus is associated with any tumor. We will examine EBV-related viral signals (DNA sequence and RNA expression) on canine oral tumors, nasal tumors, and lymphomas. In addition, we will proceed with molecular cloning of EBV-related viral sequences and culturing tumor cells. Finally, association between canine EBV-related virus and any tumors will be determined by statistical analysis

Studies on the Canine Ebv-Like Virus

EB病毒(Epstein-Barr virus, EBV)普遍感染人類，可引起傳染性單核球增生症(infectious mononucleosis)，並與多種腫瘤及紅斑性狼瘡(systemic lupus erythematosus, SLE)有關聯。EBV在分類上屬於γ-.疹病毒(gammaherpesvirus, γ-HV)亞科之隱淋巴球病毒(Lymphocryptovirus, LCV)屬，是已知感染人類之唯一的LCV屬別病毒。我們最近研究證明犬普遍曾受到與EBV極相近病毒感染，許多犬具有特異抗體識別EBV的胸腺嘧啶激.(thymidine kinase, TK)、EBV的DNA結合蛋白(EBV-encoded DNA binding protein, DBP)、與EBV的DNA聚合.(EBV-specific DNA polymerase, EDP)，在一些犬隻白血球發現EBV病毒特有BamHI W 片段之DNA序列，以及發現有些細胞強烈表現與EBV潛伏感染有關的EBV特異RNA (EBV-encoded RNA, EBER)。我們的研究為首篇犬感染γ-HV之研究報導，且為首度發現LCV屬病毒有可能感染非靈長類動物。更值得注意的是，此研究開啟了將來後續探討EBV是否會在人與犬間傳播之重要研究議題，以及利用犬做為EBV感染之動物模式之可能性。在此二年研究計畫中，我們擬繼續深入瞭解是否有更多EBV相關基因序列，特別是與細胞基因同源之EBV-encoded cellular homologues，乃至於完整的病毒基因體(complete genome)存在於犬白血球，探索所存在的EBV相關基因與犬細胞之交互作用，以及最終建立感染EBV相關病毒之犬細胞株

Studies of Zeranol-Related Mycotoxin in Rice and Soybean-Identification of Structure, Estrogenic Activities and Epigenetic Alterations

Zeranol(Z)是一種具雌性素活性的非類固醇類黴菌毒素衍生物，亦為環境內分泌干擾素之一。此毒素可由Fusarium 屬黴菌污染穀物所產生，但在米之文獻極少。Z 為FDA 核准使用於牛隻的生長促進劑，實驗顯示，以Z 促進生長的牛肉萃取液，對乳房組織培養細胞具有穩定的雌性素生物活性，且這種活性無法經加熱去除。因此食用含有Z 的食品對人類的健康，尤其是對雌性激素敏感的器官，具有極大的潛在威脅。長期食用含有低量Z 的肉品長久以來即被認為與乳癌的發生有關連。流行病學研究並顯示，日裔移民赴美後乳癌發生比例即顯著上升，可見乳癌之發生除遺傳因子外，飲食之影響亦極重要。台灣人進食牛肉的量遠不及主食米，因此若發霉米中亦有Z 之活性，則臺灣人民暴露在Z 的情形將更形嚴重。我們研究發現，自然發霉之米萃取液中可分離到具Z 免疫活性的物質，電化學層析顯示其可能為Z、β-Zearalanol(產生率為50-500 μg/g米或穀類)，以及與Z 結構並不相似之成份。此結果不同於僅有文獻中米可能產生zearalenone 及zearalenol 之報告(已被接受發表於2007 世界毒理學年會)。可見在亞熱帶台灣可能有不同的Fusarium 屬或者其他黴菌在米作用。此一新發現促使我們欲進一步研究食米中黴菌毒素與乳癌間的關係。我們提出如下假說:米及豆類食品為亞洲人民攝入Z 免疫活性物之重要來源，且這些具動情素活性之Z 免疫活性物可導致對人體健康，尤其是具高動情素受器的器官及導致乳癌發生之不良影響。因此，本研究的目的即在探討台灣本土及進口米類、豆類產品及飼料中含Z 的比例及含量，並鑑定出此一Z 活性物之結構成份及產生黴菌。此外，亦將探討此物質之雌性素活性及其與乳癌相關分子標記PTPγ間之關係。除了應用已建立的組織細胞培養模式，亦將由合作對像以動物實驗驗證食用發霉米與乳癌發生在基因內及基因外(epigenetic)表現之關係

rTS beta as a Novel 5-fluorouracil Resistance Marker of Colorectal Cancer: A Preliminary Study

Introduction: Colorectal cancer is the most common form of malignancy in Taiwan and the third leading cause of death from cancer, preceded only by lung and hepatic cancers. Colorectal cancer is typically treated by surgical intervention and/or chemotherapy and radiotherapy, if necessary. To date, 5-fluorouracil (5-FU) is the most commonly used anti-cancer chemotherapy drug. However, patients commonly experience resistance to the drug therefore limiting its efficiency. In this study, we measured the expression of rTS beta in human colon cancer as a novel 5-FU resistance marker. Materials and Methods: We collected 172 colon cancer samples from 4 different hospitals (including 21 pairs of colon cancer biopsies and 151 pathologic slides of colon cancer). In vitro, we measured the cytotoxicity of 5-FU and 5-FU plus leucovorin in H630 and H630-1 colon cancer cell lines. Results: The results revealed that rTS beta was expressed in 115 (66.9 %) pathology samples and that tumour expression was higher than in corresponding normal tissue. Survival rates of up to 5 years following treatment was significantly higher for patients without rTS beta expression than for those with rTS beta expression (P = 0.0023). In vitro, H630-1 (with rTS beta overexpression) had significantly higher IC(50) of 5-FU than did H630. IC(50) of 5-FU decreased when leucovorin was added. Conclusions: Results indicate a close relationship between rTS beta expression and resistance to the drug 5-FU in human colorectal cancer. These results provide further evidence for rTS beta expression as a novel 5-FU resistance marker of colorectal cancer