在現今的癌症治療主要仰賴化學治療，化學治療大多會伴隨許多發炎、噁心等副作用。化學治療會造成粒線體的變異使粒線體中Reactive oxygen species （ROS）升高以及粒線體中心磷脂的變異進而促使發炎反應產生。發炎反應的過程中會產生許多由二十個碳所組成的類花生酸（Eicosanoids），促使細胞發炎。在先前實驗中，利用化療藥物來抑制癌細胞的生長週期，使癌細胞停滯在不同細胞週期。發現當細胞停滯在不同細胞週期時，粒線體產生變異並導致心磷脂總量會呈現倍數的累積以及其碳鏈數的改變。我們利用NAO染色配合共軛聚焦顯微鏡檢測經化療藥物刺激過後的細胞，發現心磷脂在細胞核的周圍大量累積。當以海馬能量測定儀測定粒線體活性，發現經過化療藥物刺激過後，在心磷脂濃度變高的情況下，亦造成粒線體運作效率提高。在化療中常會伴隨著許多的發炎反應，因此本實驗利用極致高效能液相質譜儀（UPLC/Q-TOF/MS）來分析化療藥物刺激過後，細胞中的類花生酸反應。發現測得的常見類花生酸PGE2、PGD2、LTB4及5-HETE皆在化療藥物調控後，有呈現上升的趨勢。利用RT-qPCR檢測後發現，促進發炎反應蛋白COX及LOX的基因表達量也相對上升，因此確定化療中的細胞呈現發炎的狀態。另外，調控心磷脂的相關蛋白在因MMC以及MTX引發之細胞週期而產生不同的變化，並間接影響粒線體的融合及分裂。 Chemotherapy is a main treatment for cancer. Most of the chemotherapies are accompanied with many side effects, such as inflammation, nausea. Chemotherapy can induce mitochondria to make Reactive oxygen species （ROS） and of the accumulation and oxidation of cardiolipin in mitochondria can promote formation of inflammasome. Inflammation will produce a lot of 20-carbon eicosanoids. In previous experiments, chemotherapy drugs inhibited the cell cycle and the cardiolipin in the mitochondria accumulated and exhibited changes in CL species. After chemotherapy drug treatments, the cells were stained by NAO and then observed by the conjugated microscopy. Cardiolipin was significantly accumulated surrounding nucleus. The analysis by seahorse XFe extracellular flux analyzer showed that the increased CL concentration also stimulate the mitochondrial activity. We further performed UPLC/Q-TOF/MS analysis on eicosanoid and found that the increases of most eisosanoids after chemotherapy. The pro-inflammatory genes were up-regulated, which was detected by RT-qPCR. Therefore, we confirmed that the chemotherapy treated cells were in an inflamed state. In addition, MMC and MTX triggered cell cycle arrest induced gene expression changes on the cardiolipin regulation related genes, and also indirectly affect the gene expression of the mitochondrial fusion and division regulating genes.
