肺腺癌為非小細胞型肺癌(non-small cell lung cancer)的一種，是台灣目前最常發生的惡性腫瘤之一。肺腺癌早期症狀不明顯且不易察覺，但末期常併發遠端轉移及惡性肋膜腔積水。臨床上肺癌末期病人若併發惡性肋膜腔積水，通常被認為預候較差及有較高的死亡率。目前針對併發惡性肋膜腔積水的病人尚無較好的治療方式，因而尋求有效的治療策略是極必須的。腫瘤的生長及轉移需要血管新生來提供其氧氣及養分，乃至於向外轉移的路徑，因此目前有許多研究致力於阻斷腫瘤生長中所需的血管新生作用以達到治療癌症的目的。除了抑制腫瘤的生長，抗血管新生治療也可藉降低血管通透性以減緩肋膜腔積水的生成。因此本實驗嘗試以重組腺病毒攜帶血管阻斷素(angiostatin)作基因治療，來針對併發惡性肋膜腔積水的肺腺癌作治療。在基因治療中，治療基因的表現效率是關係治療成功與否很重要的一環。一般在基因治療中常使用人類巨細胞病毒啟動子(cytomegalovirus major immediate-early promoter)這個被認為可持續大量表現目標基因的啟動子來表現治療基因。在針對人類巨細胞病毒啟動子的研究中指出，在其DNA序列上具有四個nuclear factor-κB (NF-κB)的結合位置，且啟動子的轉錄活性會在細胞受到壓力時上升。因此，本實驗擬利用肺癌治療中常使用的化療藥物cisplatin與用人類巨細胞病毒啟動子表現血管阻斷素的重組病毒作合併治療，希望藉cisplatin產生的細胞毒殺壓力使人類巨細胞病毒啟動子的轉錄效率提高，使抗血管新生基因治療有更好的效果。在細胞實驗中我們證實cisplatin的確可使人類巨細胞病毒啟動子轉錄效率提高，使得治療基因血管阻斷素的表現量提高。為進一步證實此治療策略的效果，我們建立了以胸腔注射路易斯氏肺癌(Lewis lung carcinoma, LL2)細胞株，使其在肋膜腔中生長，進一步產生惡性肋膜腔積水的動物模式來評估治療效果。動物實驗的結果證實，合併cisplatin及重組腺病毒的基因治療，可有效減緩肋膜腔中腫瘤的生長並降低惡性肋膜腔積水的生成。由結果顯示，利用化療藥物強化治療基因表現的治療策略或許可以成為具潛力的癌症治療方法。

　Lung adenocarcinoma, one cell type of non-small cell lung cancer (NSCLC), is one of the most common human malignant tumors. The end stage of lung adenocarcinoma is often associated with distant metastasis and formation of malignant pleural effusion (PE). Malignant PE has consistently been shown to indicate a poor prognosis in advanced lung cancer patients, and associated with high morbidity and mortality. Thus, an effective therapeutic modality is urgently needed. Angiogenesis is necessary to supply oxygen and nutrition for tumor growth in both primary and metastatic sites. Increased vascular permeability and leakage resulting from angiogenesis play a pivotal role in the development of malignant PE. Therefore, blockade of angiogenesis has been expected to prevent not only tumor growth but also PE formation, and thus improving the prognosis of patients with NSCLC. Although the human cytomegalovirus (CMV) major immediate-early promoter has been regarded as a constitutive promoter, there are four nuclear factor-κB (NF-κB) response elements on CMV IE promoter, and cellular stress responses are known to enhance transcription from this promoter. It has been reported that induction of genotoxic stress by cisplatin, a commonly used chemotherapeutic agent for lung cancer, enhances NF-κB activity. Therefore, cisplatin treatment in combination with adenoviral vector encoding anti-angiogenic molecules driven by the CMV IE promoter is expected to have synergistic therapeutic efficacy against lung adenocarcinoma. In this study, we used Lewis lung carcinoma (LL2) cells to establish an orthotopic lung adenocarcinoma in C57BL/6 mice. LL2 cells grew in the pleural cavity, invaded contiguous structures, including diaphragm, mediastinum, pericardium, and lung parenchyma, and produced PE in the mice. In this model, when tumor-bearing mice were treated with cisplatin, smaller tumor nodules and fewer or no PE was found, suggesting that this model can be used to evaluate various treatment regimens. Cisplatin treatment enhanced CMV IE promoter activity probably due to NF-κB in LL2 cells, as determined by a reporter assay. Cisplatin treatment in conjunction with AdAST, an adenoviral vector encoding angiostatin, significantly reduced tumor weight and decreased PE volume in mice bearing orthotopic LL2 lung adenocarcinoma. Taken together, these results suggest that angiostatin delivered by adenoviral vector in combination with cisplatin may have therapeutic potential for the treatment of malignant PE associated lung adenocarcinoma.

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