以蛋白質體學探討ATP合成酶抑制劑在肺癌異種移植模式的作用

Proteomic analysis reveals the effect of targeting ectopic ATP synthase by citreoviridin in lung cancer xenograft model

吳怡萱(Yi-Hsuan Wu)

指導教授 ： 阮雪芬

國立臺灣大學/生命科學院/分子與細胞生物學研究所/碩士(2012年)

https://doi.org/10.6342/NTU.2012.10954

摘要

先前的研究發現ATP合成酶在多種癌細胞的細胞膜上表現，使用ATP合成酶抑制劑黃綠青黴素(citreoviridin)可以在不影響正常細胞下，而抑制肺癌細胞增生與腫瘤生長。由於在體內抑制表面型ATP合成酶的整體影響仍未清楚，因此在本研究中我們以相對和絕對定量的等量異位標籤(isobaric tags for relative and absolute quantitation, iTRAQ)定量蛋白質體學來分析citreoviridin在肺癌異種移植(xenograft)模式中的調控。我們以良好的再現性鑑定並定量了2,659個蛋白質，並且以它們在控制組與citreoviridin處理組的相對表現量挑選出表現量有差異的141個蛋白質。使用生物資訊學方法加以分析表現量有差異的蛋白質，結果發現在肺癌中citreoviridin會改變與葡萄糖代謝相關酵素的表現。參與糖質新生(gluconeogenesis)與葡萄糖儲存的酵素表現量增加，顯示citreoviridin可能藉由降低提供給合成巨分子所需的糖解作用中間產物來達到抑制癌細胞增生的效果。本研究以蛋白質體學探討citreoviridin抗肺癌作用可能影響代謝的功能，根據此研究結果，我們希望能有助於深入了解代謝與腫瘤生成的關係，並提供肺癌治療的策略方向。

關鍵字

蛋白質體學 ； 相對和絕對定量的等量異位標籤(iTRAQ) ； 表面型ATP合成酶肺癌 ； 糖質新生

並列摘要

ATP synthase is present on the plasma membrane of several types of cancer cells. Citreoviridin, an ATP synthase inhibitor, selectively suppresses the proliferation and tumor growth of lung cancer without affecting normal cells. However, the global effects of targeting ectopic ATP synthase in vivo have not been well defined. In this study, we performed quantitative proteomic analysis using isobaric tags for relative and absolute quantitation (iTRAQ) and provided a comprehensive insight into the complicated regulation by citreoviridin in lung cancer xenograft model. With high reproducibility of the quantitation, we obtained quantitative proteomic profiling with 2,659 proteins identified. Bioinformatics analysis of the 141 differentially expressed proteins selected by their relative abundance revealed that citreoviridin induces alterations in the expression of glucose metabolism-related enzymes in lung cancer. The up-regulation of enzymes involved in gluconeogenesis and storage of glucose indicated that citreoviridin may reduce the glycolytic intermediates for macromolecule synthesis and inhibit cell proliferation. The results provide metabolic aspects from comprehensive proteomics for the antitumorigenic effect of citreoviridin in lung cancer, which may lead to a better understanding of the links between metabolism and tumorigenesis in cancer therapy.

並列關鍵字

proteomics ； iTRAQ ； ectopic ATP synthase ； lung cancer ； gluconeogenesis

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