Abstract
Cellular volatile organic compounds (VOCs) are unique compounds whose metabolic pathways remain enigmatic. To elucidate their metabolism, we investigated the VOCs of lung cancer A549 and 2 non-cancer lung cells (HLB; HBEpC). Neutral sugars and lactate in the medium were measured by colorimetric assay. VOCs were enriched by monotrap and profiled by GC-MS. To investigate the enzymes that change VOC metabolism in cells, we conducted ALDH activity assays and qPCR. ROS (reactive oxygen species) assays were conducted to assess oxidation stress. The colorimetric assay showed that especially A549 and HLB took up sugars from the medium and rapidly secreted lactate into the medium. The VOC profile (GC-MS) revealed a trans-2-hexenol increase, especially in A549 lung cancer cells. This is a novel lipid peroxidation product from animal cells. Based on the absolute quantification data, trans-2-hexenol increased in parallel with number of A549 cancer cells incubated. The qPCR data implies that ADH1c potentially plays an important role in the conversion into trans-2-hexenol.
Highlights
The potential relationship between Volatile organic compounds (VOC) emitted by the human body and malignant diseases has been investigated since Hippocrates’ time (Di Francesco et al, 2005)
Based on the VOC profile results, we focus on trans-2hexenol metabolism
Data obtained by the conventional phenol-sulfuric acid assay indicated that the neutral sugar concentration of all cell culture media gradually decreased over time in the 2D cell culture (Figure 2A)
Summary
The potential relationship between VOCs (volatile organic compounds) emitted by the human body and malignant diseases (e.g., cancer) has been investigated since Hippocrates’ time (Di Francesco et al, 2005). A gap remains between disease detection and volatile compound analysis. One reason is that the relationship between pathology and the metabolic pathways producing VOC is not fully understood. The mechanism of biological VOC synthesis in cells is enigmatic because many studies on such metabolites are restricted to bacteria, algae, and fungi (Ruffing, 2013). The enzymes associated with profiled cell VOCs have not always been identified (e.g., hydrocarbons, alcohols, alkenes). The goal of our study is to elucidate the VOC pathway which can potentially be marker for cancer. VOC analysis by GC-MS profiles (Buszewski et al, 2012) is certainly useful for assessing VOCs as biomarkers, but there is some room for improvement in VOC sampling
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