2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38574Non-small cell lung cancer (NSCLC) accounts for 85% of lung cancer patients and 70% of female NSCLC in Taiwan. They are usually associated with poor prognosis, more than one half of NSCLC patients have advanced disease at diagnosis and beyond the limits of surgery. In spite of new treatments, the overall five-year survival rate remains about 14% and most patients present with advanced disease. Obviously, novel therapeutic strategies to improve efficacy in according with safety are urgently needed. The survival in patients with advanced stage NSCLC can be prolonged with chemotherapy, but drug resistance remains the important issue of chemotherapy. Multidrug resistance gene (MDR-1) mediated by P-glycoprotein has been studied in lung cancer but evidences have been gathered that MDR-1 may be not responsible for drug resistance in lung cancer chemotherapy. We have reported that docetaxel (TaxotereR, DOC) enhances p53 but not tubulin in A549 cells; whereas, DOC enhances tubulin in H1299 cell that p53 expression is absence. Therefore, p53 is associated with the sensitivity of DOC in lung cancer cell lines. We though the mechanism of DOC resistance might be regulated differently in cells contain either wild type p53 or defected p53. The long-term goal of this proposal is to study the mechanism of drug resistance by DOC, a generally applied second line chemotherapeutic drug for NSCLC. Moreover, we also intend to develop an anti-resistance therapy to efficiently improve the efficacy of chemotherapy. Our first year』s goals are to examine the roles of p53, tubulin and MDR-1 in DOC-resistance by using the DOC-resistance NSCLC cell lines and determine the gene interaction profile using DNA microarray. After the DOC inducing genes are identified, further characterization will be applied to confirm the association of DOC resistance through out this study. The second year』s goals are to determine the DOC-response sequences using the DNA upstream regulatory sequences of p53 and beta-tubulin by luciferase reporter assay, which may reveal the signaling pathway in DOC regulation. To understand the mechanism of upregulation of p53 and tubulin by DOC should provide future benefit to reverse DOC resistance. The mechanisms that cause chemoresistance of DOC in NSCLC patients have yet to be clearly elucidated. Accumulating evidence suggests that Clusterin (Clu, a secreted protein) is an anti-apoptotic protein associated with both cancer therapy resistance and tumor development. Overexpression of clusterin was reported to be closely related to disease progression and recurrence in patients with several cancers. Therefore, the third year』s goals are to explore the potential anti-apoptotic function of clusterin in NSCLC cells and to examine the role of clusterin associated chemoresistance. Our ultimate goal is to prolong survival, minimizing adverse events, control symptoms, and improve the quality of life for patients with lung cancer in Taiwan.近年台灣肺癌病患以罹患肺腺癌的比例最高，特別是女性患者中，比例高達七成， 肺腺癌已成為婦女的重要死因。而非小細胞肺癌的治療情形則不甚理想，就算是使用新 的化學治療藥物，五年存活率也只有14%，尤其是其抗藥性問題是目前臨床治療的最大 困擾。根據臨床經驗，大部份非小細胞肺癌患者由於藥物無法有效控制，因此從診斷出 至死亡一般不會超過一年。有關非小細胞肺癌的抗藥性機轉已有許多報告。其中 Multi-Drug Resistance (MDR-1)基因是被研究較多之機轉，事實上尚有其它機轉正逐 漸被發現出來，但非小細胞肺癌的抗藥性是否確實因上述機轉或其組合所引起尚未被完 全肯定，仍有待進一步的證實。故非小細胞肺癌之抗藥性機轉目前仍是肺癌學術研究的 主題。因此我們在此設法突破癌細胞之抗藥性，才會對肺癌的化學治療提供有效改善治 療的結果。 我們計畫以臨床上普遍應用於第二線治療非小細胞肺癌之歐洲紫杉醇衍生物 (Docetaxel) (商品名為Taxotere 剋癌易)做為研究非小細胞肺癌之抗藥性機轉。 Docetaxel (DOC)的毒性機轉是促進微小管次體的聚合，並抑制微小管次體的去聚合作 用，達到中斷有絲分裂的藥效，最後造成癌細胞不正常的細胞漿移動而死亡。之前，我 們報導過肺癌細胞株對DOC 之敏感性與抑癌基因p53 和tubulin 有關，對沒有抑癌基因 p53 的H1299 細胞使用DOC 會造成tubulin 蛋白質的增加，但是對具有正常抑癌基因p53 的A549 細胞只見到抑癌基因p53 的表達增加卻見不到tubulin 蛋白質的增加。反之， 在H1299 細胞中表達p53 和在A549 細胞抑制p53 則會對DOC 的使用在p53 和tubulin 的表達上得到與之前相反的結果。因此我們設定長期的目標是研究具有正常抑癌基因 p53 和沒有正常抑癌基因p53 的肺癌細胞，對DOC 產生抗藥性的調控機轉是否不同。並 且設法減低抗藥性的產生。因為在肺癌腫瘤中超過半數是會伴隨有抑癌基因p53 的缺 失，所以針對抑癌基因p53 和tubulin 來研究因使用DOC 而產生的抗藥性是重要的。 因此本計畫之第一年目標為使用具有DOC 之抗藥性的肺癌細胞株做為材料，並且找 出p53、tubulin 和MDR-1 與DOC 之抗藥性的關係，同時，利用DNA 微探針陣列比對會 被DOC 轉錄活化的基因，並且逐步地在計畫期間進一步比對和驗證其與抗藥性的關聯。 第二年目標為研究DOC 如何控制p53 和tubulin 的基因轉錄活性，因而產生抗藥性。我 們將會把p53 和tubulin 的基因轉錄調控區分離出來尋找它們對DOC 的反應序列。事實 上大部分的DOC 引起之基因訊息的傳遞路徑仍未完全建立，身體細胞內基因調控機制亦 尚不清楚，仍有許多因子尚未被發現而待進一步確立。先前針對癌症組織所進行的研究 發現，Clusterin (為一種分泌性蛋白質)可大量出現在包括前列腺、腎臟、膀胱、肺臟、 卵巢等癌細胞中。人體的健康細胞和疾病細胞均會利用Clusterin 作為盾牌，抵擋外來 攻擊。本計畫之第三年目標為研究Clusterin 在非小細胞肺癌的作用，及它如何保護癌 細胞免受如化學治療等療法的攻擊。利用抑制Clusterin 基因表達方法來強化現有癌症 治療藥物的效果。因此我們對於Clusterin 這個蛋白質是否與肺癌的抗藥性有關亦將深 入探討。並且計畫將抗凋亡的基因(Bcl-2, NFkB)加以抑制來降低對DOC 之抗藥性，增 加化學治療的效果，改善病患的生活品質。Project