Zhou Jijuan, Ge Dianlong, Liu Yue, Chu Yajing, Zheng Xiangxue, Lu Yan, Chu Yannan
Anhui Province Key Laboratory of Medical Physics and Technology, Institute of Health and Medical Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, Hefei 230031, P. R. China.
University of Science and Technology of China, Hefei 230026, P. R. China.
Anal Chem. 2024 Dec 31;96(52):20398-20405. doi: 10.1021/acs.analchem.4c03894. Epub 2024 Dec 18.
Cellular volatile organic compound (VOC) detection is crucial for studying lung cancer biomarkers. However, the reported VOC biomarkers from the same cell line seem to be inconsistent across different research groups. It is possibly related to the variation in culture media, and the result obtained by a conventional single medium approach (SMA) depends on what medium is used in the cell experiment. This study proposes a multiple media approach (MMA) to investigate reproducible characteristic VOCs of lung cancer cells. Using solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) in combination with untargeted analysis, we conducted two independently repetitive experiments to compare lung cancer cells (A549) and normal lung cells (BEAS-2B) under three culture media conditions (RPMI 1640, DMEM, and Ham's F12). Both experiments indicated that, compared with 62-96 VOCs obtained by the SMA, only two VOCs (3-methyl-1-butanol and 2-methyl-1-butanol) were reproducibly achieved by the MMA. Moreover, their concentrations were significantly lower in lung cancer cells than in normal cells. Further targeted analysis confirmed the downregulation trend of both VOCs in subcutaneous and primary tumor tissues from the lung cancer model mouse. The present work demonstrated that the MMA cell experiment, just like the multicenter trials for cell lines, can facilitate the discovery of reproducible characteristic VOCs. This provides a cellular experimental basis and scientific evidence for lung cancer biomarker investigation and even breath biopsy technique development.
细胞挥发性有机化合物（VOC）检测对于研究肺癌生物标志物至关重要。然而，来自同一细胞系的已报道VOC生物标志物在不同研究组之间似乎并不一致。这可能与培养基的差异有关，并且传统单一培养基方法（SMA）所获得的结果取决于细胞实验中使用的是何种培养基。本研究提出了一种多培养基方法（MMA）来研究肺癌细胞可重复的特征性VOC。我们使用固相微萃取-气相色谱-质谱联用（SPME-GC-MS）结合非靶向分析，在三种培养基条件（RPMI 1640、DMEM和Ham's F12）下对肺癌细胞（A549）和正常肺细胞（BEAS-2B）进行了两次独立的重复实验。两个实验均表明，与SMA获得的62 - 96种VOC相比，MMA仅可重复获得两种VOC（3-甲基-1-丁醇和2-甲基-1-丁醇）。此外，它们在肺癌细胞中的浓度显著低于正常细胞。进一步的靶向分析证实了这两种VOC在肺癌模型小鼠的皮下和原发肿瘤组织中均呈下调趋势。目前的工作表明，MMA细胞实验与细胞系的多中心试验一样，能够促进可重复特征性VOC的发现。这为肺癌生物标志物研究乃至呼气活检技术的发展提供了细胞实验基础和科学依据。
