Yang Zhen, Zhang Qiuning, Luo Hongtao, Shao Lihua, Liu Ruifeng, Kong Yarong, Zhao Xueshan, Geng Yichao, Li Chengcheng, Wang Xiaohu
The Basic Medical College of Lanzhou University, Lanzhou, China.
Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, China.
Front Oncol. 2021 Mar 2;10:601620. doi: 10.3389/fonc.2020.601620. eCollection 2020.
To analyze the effect of carbon ion (C) radiation may induce bystander effect on A549 cell metastasis and metabonomics.
A549 cell was irradiated with carbon ion to establish the clone survival model and the transwell matrix assay was applied to measure the effect of carbon ion on cell viability, migration, and invasion, respectively. Normal human embryonic lung fibroblasts (WI-38) were irradiated with carbon ions of 0 and 2 Gy and then transferred to A549 cell co-culture medium for 24 h. The migration and invasion of A549 cells were detected by the Transwell chamber. The analysis of metabonomic information in transfer medium by liquid phase mass spectrometry (LC-MS), The differential molecules were obtained by principal pomponent analysis (PCA) and the target proteins of significant differences ( = 1.7 × 10) obtained by combining with the STICH database. KEGG pathway was used to analyze the enrichment of the target protein pathway.
Compared with 0 Gy, the colony formation, migration, and invasion of A549 cells were significantly inhibited by carbon ion 2 and 4 Gy irradiation, while the inhibitory effect was not significant after 1 Gy irradiation. Compared with 0 Gy, the culture medium 24 h after carbon ion 2 Gy irradiation significantly inhibited the metastasis of tumor cells ( = 0.03). LC-MS analysis showed that 23 differential metabolites were obtained in the cell culture medium 24 h after carbon ion 0 and 2 Gy irradiation (9 up-regulated and 14 down-regulated). Among them, two were up-regulated and two down-regulated ( = 2.9 × 10). 41 target proteins were corresponding to these four differential molecules. Through the analysis of the KEGG signal pathway, it was found that these target molecules were mainly enriched in purine metabolism, tyrosine metabolism, cysteine and methionine metabolism, peroxisome, and carbon metabolism. Neuroactive ligand-receptor interaction, calcium signaling pathway, arachidonic acid metabolism, and Fc epsilon RI signaling pathway.
The bystander effect induced by 2 Gy carbon ion radiation inhibits the metastasis of tumor cells, which indicates that carbon ions may change the metabolites of irradiated cells, so that it may indirectly affect the metabolism of tumor cell growth microenvironment, thus inhibiting the metastasis of malignant tumor cells.
分析碳离子（C）辐射诱导的旁观者效应对A549细胞转移及代谢组学的影响。
用碳离子辐照A549细胞建立克隆存活模型，应用Transwell基质实验分别检测碳离子对细胞活力、迁移和侵袭的影响。用0 Gy和2 Gy的碳离子辐照正常人胚肺成纤维细胞（WI-38），然后转移至A549细胞共培养基中培养24 h。用Transwell小室检测A549细胞的迁移和侵袭。用液相质谱（LC-MS）分析转移培养基中的代谢组学信息，通过主成分分析（PCA）获得差异分子，并结合STICH数据库获得差异显著（P = 1.7×10）的靶蛋白。用KEGG通路分析靶蛋白通路的富集情况。
与0 Gy相比，2 Gy和4 Gy碳离子辐照显著抑制A549细胞的集落形成、迁移和侵袭，而1 Gy辐照后抑制作用不显著。与0 Gy相比，2 Gy碳离子辐照24 h后的培养基显著抑制肿瘤细胞的转移（P = 0.??03）。LC-MS分析显示，0 Gy和2 Gy碳离子辐照24 h后的细胞培养基中获得23种差异代谢物（9种上调，14种下调）。其中，4种差异显著（P = 2.9×10），2种上调，2种下调。41种靶蛋白与这4种差异分子相对应。通过KEGG信号通路分析发现，这些靶分子主要富集在嘌呤代谢、酪氨酸代谢、半胱氨酸和甲硫氨酸代谢、过氧化物酶体和碳代谢、神经活性配体-受体相互作用、钙信号通路、花生四烯酸代谢和FcεRI信号通路。
2 Gy碳离子辐射诱导的旁观者效应抑制肿瘤细胞转移，这表明碳离子可能改变受照细胞的代谢物，从而间接影响肿瘤细胞生长微环境的代谢，进而抑制恶性肿瘤细胞的转移。
