Hayasaka Ryosuke, Tabata Sho, Hasebe Masako, Ikeda Satsuki, Ohnuma Sumiko, Mori Masaru, Soga Tomoyoshi, Tomita Masaru, Hirayama Akiyoshi
Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata 997-0052, Japan.
Systems Biology Program, Graduate School of Media and Governance, Keio University, Fujisawa, Kanagawa 252-0882, Japan.
Metabolites. 2021 Apr 1;11(4):215. doi: 10.3390/metabo11040215.
Extracellular vesicles (EVs) released from cancer cells contribute to various malignant phenotypes of cancer, including metastasis, cachexia, and angiogenesis. Although DNA, mRNAs, miRNAs, and proteins contained in EVs have been extensively studied, the function of metabolites in EVs remains unclear. In this study, we performed a comprehensive metabolomic analysis of pancreatic cancer cells, PANC-1, cultured under different oxygen concentrations, and small EVs (sEVs) released from them, considering the fact that hypoxia contributes to the malignant behavior of cells in pancreatic cancer, which is a poorly diagnosed cancer. sEVs were collected by ultracentrifugation, and hydrophilic metabolites were analyzed using capillary ion chromatography-mass spectrometry and liquid chromatography-mass spectrometry, and lipids were analyzed by supercritical fluid chromatography-tandem mass spectrometry. A total of 140 hydrophilic metabolites and 494 lipids were detected in sEVs, and their profiles were different from those in cells. In addition, the metabolomic profile of sEVs was observed to change under hypoxic stress, and an increase in metabolites involved in angiogenesis was also detected. We reveal the hallmark of the metabolites contained in sEVs and the effect of tumor hypoxia on their profiles, which may help in understanding EV-mediated cancer malignancy.
癌细胞释放的细胞外囊泡（EVs）会导致癌症的各种恶性表型，包括转移、恶病质和血管生成。尽管对EVs中所含的DNA、mRNA、miRNA和蛋白质已进行了广泛研究，但EVs中代谢物的功能仍不清楚。在本研究中，鉴于缺氧会导致胰腺癌（一种难以诊断的癌症）细胞的恶性行为，我们对在不同氧浓度下培养的胰腺癌细胞PANC-1及其释放的小EVs（sEVs）进行了全面的代谢组学分析。通过超速离心收集sEVs，使用毛细管离子色谱-质谱联用仪和液相色谱-质谱联用仪分析亲水性代谢物，通过超临界流体色谱-串联质谱分析脂质。在sEVs中总共检测到140种亲水性代谢物和494种脂质，它们的谱图与细胞中的不同。此外，观察到sEVs的代谢组学谱图在缺氧应激下会发生变化，并且还检测到参与血管生成的代谢物增加。我们揭示了sEVs中所含代谢物的特征以及肿瘤缺氧对其谱图的影响，这可能有助于理解EV介导的癌症恶性行为。
