2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38529Genome-wide analysis showed that HPV-positive lung adenocarcinomas (AD) had higher allelic imbalance (AI) at some loci than those of HPV-negative AD. Among these loci, a higher AI of DDX3 located at Xp11.4 in HPV-infected AD was observed as compared with HPV-noninfected AD. RS-PCR data showed that Xp11.4 was a integrated site of HPV DNA in lung AD. We thus speculated that DDX3 AI could be caused by HPV DNA integration. Immunohistochemistry analysis showed that a higher prevalent of DDX3 immunostaining negative was found in AD with DDX3 AI, suggesting that DDX3 AI could reduce the expression of DDX3 proteins in HPV-infected lung AD. A recent report indicated that p21 transcription was upregulated by DDX3 by p53 independent pathway. Real-time RT-PCR data showed that p21 mRNA level in DDX3-negative AD was lower than those with DDX3-postive AD. We thus suggest that reduced DDX3 by HPV integration may alter p21 transcription to involve in lung tumorigenesis. To further verify whether HPV infection was linked with DDX3 aberration in genome level, DDX3 DNA copy number of HPV16 E6-positive TL-1 and HPV16 E6-negative TL-4 lung adenocarcinoma cells, which were established from patient’s pleural effusions, were evaluated by real-time PCR. Our data showed that DDX3 DNA copy number in E6-positive TL-1 cells was 0.6-fold of E6-negative TL-4 cells. Additionally, DDX3 DNA copy number in TL-4 cells was not changed by the transfection of HPV16 E6. These observations suggest that a lower DDX3 DNA copy number in TL-1 cells could be due to HPV16 DNA integration, not by E6. In conclusion, DDX3 alteration by HPV16/18 DNA integration at Xp11.4 may at least in part involved in HPV-associated lung tumorigenesis.本研究利用Genome-wide 分析發現，HPV感染的肺腺癌在許多宿主染色體位置發生基因不穩定，其頻率遠高於沒有HPV感染的肺腺癌，其中包含了位於Xp11.4的DDX3基因。RS-PCR的結果HPV感染之肺腫瘤確實發現Xp11.4是HPV16 DNA嵌入的位置。因此推測DDX3的基因不穩定，可能會經由HPV DNA 嵌入該位置所致。首先利用免疫組織染色分析DDX3蛋白是否與基因不穩定相關？結果發現DDX3基因不穩定腫瘤之DDX3的蛋白表現顯著低於沒有發生DDX3基因不穩定之腫瘤。過去研究指出DDX3會不經由p53路徑調控p21Waf/Cip1之轉錄活性。Real-time RT-PCR和免疫組織化學分析的結果都顯示在DDX3低表現之腫瘤較DDX3表現高者之p21Waf/Cip1 mRNA 和其蛋白有較低的表現。因此本研究推測HPV DNA嵌入可能會造成DDX3基因不穩定，而降低DDX3基因的表現，進而抑制p21Waf1/Cip1之轉錄活性。本研究以real-time PCR分析同一肺癌病患胸水培養出來之HPV16感染之肺腺癌細胞 (TL-1)以及沒有HPV感染之TL-4細胞之DDX3 DNA複製數目（copy number），結果發現有HPV16感染之TL-1之DNA複製數目，僅有TL-4細胞的0.6倍。若將HPV16 E6轉殖入TL-4細胞，結果DDX3 DNA的複製數目並沒有改變。由以上結果推測HPV16 E6的表現，並不會影響DDX3 DNA的複製數目，有可能主要是經由HPV16 DNA嵌入宿主染色體而引起DDX3 DNA缺失。因此HPV感染引起肺腫瘤，可能是經由HPV DNA之嵌入而造成DDX3去活化。因此會不經由p53路徑而抑制p21Waf1/Cip1的表現而參與肺腫瘤化。HPVDDX3肺癌肺腺癌HPV1618DDX3lung cancerProject