人類乳突病毒和台灣女性肺癌之相關性研究

中山醫學大學機構典藏 CSMUIR > 醫學院 > 醫學分子毒理學研究所 > 研究計劃 > Item 310902500/3336

Please use this identifier to cite or link to this item: https://ir.csmu.edu.tw:8080/ir/handle/310902500/3336

Title:	人類乳突病毒和台灣女性肺癌之相關性研究 The Association of Hpv with Lung Cancer in Taiwanese Women
Authors:	鄭雅文;吳德威 Cheng, Ya-Wen;Wu, De-Wei
Contributors:	中山醫學院毒理學研究所
Keywords:	肺癌;人類乳突病毒 Lung cancer;human papillomavirus
Date:	2004
Issue Date:	2010-12-17T03:05:21Z (UTC)
Abstract:	為了進一步篩檢HPV可能參與肺癌形成之重要基因，本年度計劃利用有及沒有轉染HPV E6 之肺癌細胞株進行cDNA microarray分析研究，本研究之初步成果主要將表現差異較大之基因依據Nees et al., 2001的研究區分為四類， Cluster I: INF-related gene，在microarray 的分析結果中此類基因在HPV+與HPV- 的腫瘤組織中表現量隨HPV 16/18 E6/E7 之表現而有抑制之現象。Cluster II：NF-κB related gene，已知NF-κB基因受 TNF-α 所調控，TNF-α 可活化 NF-κB及AP1 等轉錄因子，並進而刺激許多與發炎反應有關之基因表現，而在本研究之結果中此部份基因有58.8% 在HPV 致癌蛋白表現者有被抑制的現象，因此肺腫瘤組織中HPV 16/18 E6/E7 致癌蛋白的表現確實會影響細胞之免疫功能。Cluster III：Cell cycle related gene and DNA synthesis related gene，本研究結果顯示，此類基因在有HPV感染之肺腫瘤之表現量較沒有感染之肺腫瘤有增加的現象，這結果顯示HPV 感染確實會影響細胞週期之調控。而Cluster IV：Other，此類主要包括膜蛋白及結構蛋白等而此部份選取之基因亦有66.7%被抑制，主要影響細胞間物質交換之通道及細胞之結構，但詳細之機轉仍需進一步研究。在先前計畫之cDNA microarray的結果已獲知，HPV感染之肺腫瘤中參與細胞週期與DNA合成以及免疫相關之基因表現輿圖與E6與E7 轉形之子宮頸角質細胞(transformed cervical keritinocytes)以及肺癌細胞轉染 E6, E7之表現輿圖相似。這顯示HPV感染確實會改變肺腫瘤基因之表現，而與子宮頸角質細胞感染HPV引起轉形之過程相似。本研究室過去以RS-PCR結果發現HPV 16/18 DNA會嵌入到染色體1, 12, 17, 18和X中，顯示HPV DNA 嵌入到肺腫瘤細胞可能是隨機性的，因此欲找到HPV嵌入引起之染色體LOH，可能須要以genome-wide全面篩檢方式，較能找到參與HPV感染引起之肺腫瘤化的抑癌基因(tumor suppressor genes, TSGs)。本計畫進一步以genomic-wide 分析人類乳突病毒感染之不抽菸女性肺腺癌主要發生異質性基因座缺失 (loss of heterozygosity, LOH)之染色體區域，並希望能經由發生LOH位置，找到並確定主要參與之已知與未知之TSGs。本計劃目前已完成Chromosome 1-10的基因不穩定分析，結果發現HPV嵌入宿主細胞染色體中的可能造成下列位置發生基因不穩定，分析結果發現HPV16/18感染可能如子宮頸癌一樣，會經由嵌入宿主染色體造成基因不穩定而參與肺腫瘤之形成。本計畫之研究結果除了發現肺組織確實可偵測到HPV病毒外，並由血液中之檢測推測HPV之感染可能來自血液循環，且肺組織中感染之HPV 病毒確實會嵌入宿主DNA 中產生E6 致癌蛋白，並使宿主發生基因不穩定，而造成肺腫瘤化。本年度計畫之研究結果，將有助於了解HPV感染與肺細胞腫瘤化之相關性。 Our previous report have indicated that extremely higher frequencies of HPV 16 (60%) and 18 (73%) infections were found in female lung cancer patients as compared with male cases (24% for HPV 16, 26% for HPV 18). Therefore, we suggested that HPV 16/18 infection may be associated with female lung cancer development in Taiwan. Our recent case-control study showed that subject with HPV 16/18 infections in blood circulation had 38.52-fold of lung cancer risk of subject without HPV 16/18 DNA in blood circulation (P = 0.0004,). Thus, the presence of HPV 16/18 DNA in blood circulation may act as a risk marker of lung cancer in Taiwan. To elucidate the HPV transmission route in female lung cancer patient, palp smear and blood samples were collected from 15 female lung cancer patients for HPV 16/18 DNA detection. Our data showed that HPV 16 infection was correlated between palp smear and blood cells (p = 0.022), but the correlation was not observed in HPV 18. This result seems to support the speculation that HPV 16 may be originated from cervix and transmitted into lung through blood circulation. To understand whether HPV infection was involved in lung tumorigenesis, LCM-cDNA microarray was performed to compare differently expressed genes including INF-, NF-kB-, cell cycle and DNA synthesis-related genes. These results suggested that HPV was indeed involved in lung tumoringenesis through alteration of immune response- and cell proliferation-related gene expression and the altered expression profile was similar with those of HPV 16/E6/E7 transfected keritinocytes. HPV DNA integration in HPV infected-lung tumors were determined by RS-PCR. All of HPV infected-lung tumors with E6 mRNA expression had positive RS-PCR result. This result strongly indicated that HPV 16/18 DNA can be integrated into the chromosomes of lung tumors. Seven integration sites were found in Xp11.3-11.4 (PAC95C20)？HHHAChromosome 12 (BACRP11-478H3), Chromosome 1 (RP11-477H21), chromosome 1 (RP11-147C23), Xp22.31-22.13 PHKA2, Chromosome 18 (RP11-138E9) and Chromosome 17 (953). Among these, the most prevalence site was observed in X chromosome (3 of 7, 42.9%), followed by Chromosome 1 (2 of 7, 28.6%). Our preliminary data seemed to reveal that HPV integration in lung cancer was not random. This phenomenon was not consistent to HPV associated-cervical cancer reported previously. A higher FHIT LOH frequency was found in HPV infected cervical cancer and this due to HPV integrated into FRB fragile site in FHIT to cause LOH. On the other hand, a higher FHIT LOH frequency was also observed in smoking lung cancer as compared to nonsmoking cases. Thus, we hypothesized that HPV infected-nonsmoking female lung cancer may have high FHIT LOH frequency as same as that of non-HPV infected-smoking male lung cancer. Our results from LOH analysis using D3S1300 marker showed that FHIT LOH frequency in female lung tumors with HPV 16 infection was significantly higher than those without HPV 16 infection, not in HPV 18. Additionally, the finding was not observed in smoking and nonsmoking male lung cancer patients. This result suggested that HPV 16 involved in female lung tumorigenesis was at least in part mediated through the increase of FHIT LOH. The genome-wide experiment was investigated in HPV-infected lung tumor in this project. Until now, chromosome 1 to 10 has been completed and our preliminary data seemed to reveal that the genome instability locus found in HPV infected lung cancer was similar to that of HPV associated cervical cancer. This result indicated that HPV was indeed involved in lung tumorigenesis through genome instability by HPV DNA integration.
URI:	https://ir.csmu.edu.tw:8080/handle/310902500/3336
Appears in Collections:	[醫學分子毒理學研究所] 研究計劃

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