非小細胞肺癌幹細胞多型性與抗藥性及預後生物標記之研究

The Heterogeneity of Lung Cancer Stem Cells in Drug Resistance and Prognostic Biomarkers of Non-Small-Cell Lung Cancer

陳昭儒(Chao-Ju Chen)

指導教授 ： 劉于鵬

共同指導教授 ： 黃明賢(Ming-Shyan Huang)

高雄醫學大學/醫學院/醫學研究所博士班/博士(2020年)

摘要

非小細胞肺癌是一種相當致命的惡性腫瘤且在全世界皆有相當高的盛行率。抗藥性導致癌症復發是目前肺癌治療上遇到的最大瓶頸之一，因此持續尋找新的治療選擇，並找出可以預測疾病預後的生物標記決定治療策略，是非常重要且迫切的。過去的研究認為，間質狀態的癌細胞具有較高的抗藥性，而癌症幹細胞也是普遍被認為是造成藥物抗藥性的元兇。然而過去對於不同上皮和間質狀態的癌症幹細胞與抗藥性間的關係仍不清楚。在本研究中我們發現，上皮狀態的癌症幹細胞比間質狀態的癌症幹細胞具有更高的幹細胞特性與化療藥物抗性。在調控機轉的研究中，我們發現上皮狀態癌症幹細胞具有較高的MDFIC蛋白質表現，細胞質中的MDFIC p32蛋白質異形體會與Axin/GSK-3/β-catenin複合物進行相互作用，使β-catenin下游路徑活化而增強細胞對於藥物的抗藥性。另一方面，我們過去的研究發現TIP30可抑制肺癌幹細胞特性及轉移。因此我們使用非小細胞肺癌病患的組織微矩陣檢體，驗證MDFIC或TIP30對肺癌病人預後的預測效能，結果發現MDFIC高表現或TIP30低表現皆和非小細胞肺癌患者的不良預後有關。因此，我們的研究結果顯示MDFIC或TIP30蛋白均具有作為非小細胞肺癌病患預後生物標記的潛力。

關鍵字

肺癌 ； 癌症幹細胞 ； 上皮間質轉化 ； 抗藥性 ； 生物標記

並列摘要

Non-small-cell lung cancer (NSCLC) is a deadly malignancy with a high prevalence worldwide. The therapeutic efficacy for most anti-cancer agents is unsatisfied due to the acquirement of drug resistance. Cancer cells with mesenchymal attributes potentially display chemoresistance. Cancer stem cells (CSCs), which are intrinsically resistant to most chemotherapy agents, exhibit considerable phenotypic heterogeneity in their epithelial versus mesenchymal states. However, the drug response of CSCs in epithelial and mesenchymal states has not been completely investigated. In this study, we found that the epithelial-type CSCs displayed a higher sphere formation ability and chemoresistance than the mesenchymal-type CSCs. Gene expression profiling of the CSC and non-CSC subpopulations with distinct epithelial-to-mesenchymal transition (EMT) states showed that MyoD family inhibitor domain-containing (MDFIC) was selectively upregulated in the epithelial-type CSCs. A mechanistic study showed that MDFIC p32 isoform, which is located in cytoplasm, interacted with Axin/GSK-3/β-catenin destruction complex leading to the increase of protein stability, nuclear translocation and transcriptional activity of β-catenin. Knockdown of β-catenin reversed MDFIC-enhanced chemoresistance. On the other hand, we previously found that Tat-interacting protein 30 (TIP30) suppresses sphere formation and metastasis by inhibiting EMT in NSCLC cells. We further analyzed the association of MDFIC or TIP30 with the prognosis of NSCLC patients by using a tissue microarray. The results showed that high MDFIC expression or low TIP30 expression was associated with the poor prognosis of NSCLC patients. Accordingly, our data indicated that either MDFIC or TIP30 would be a potential prognostic biomarker of NSCLC.

並列關鍵字

Lung cancer ； cancer stem cell ； epithelial-to-mesenchymal transition ； drug resistance ； biomarker

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