Division of Pulmonology, Department of Internal Medicine, Konyang University Hospital, Daejeon, Republic of Korea.
Myunggok Medical Research Institute, College of Medicine, Konyang University, Daejeon, Republic of Korea.
Inhal Toxicol. 2020 Nov-Dec;32(13-14):487-493. doi: 10.1080/08958378.2020.1850936. Epub 2020 Dec 7.
Particulate matter (PM) has various systemic effects. We researched the effects of PM on lung epithelial cells with next generation sequencing (NGS) and validated this with quantitative real-time polymerase chain reaction (qRT-PCR). We cultured the group exposed to PM (Particulate matter less than 10 μm)-like fine dust (ERM CZ120 fine dust) at a concentration of 50 μg/mL and the untreated group for seven days in one normal lung epithelial cell line (BEAS-2B) and four lung cancer epithelial cell lines (NCI-H358, HCC-827, A549, NCI-H292). Then, we extracted the RNA from the sample and performed NGS. As a result of NGS, various gene expressions were upregulated or downregulated. Among them, we selected the gene whose mean fold change was more than doubled and changed in the same direction in all five cell lines. Based on these genes, we selected the top 10 genes, either upregulated or downregulated, to validate with the qRT-PCR. There were the four genes that matched the NGS and qRT-PCR results, all of which were upregulated genes. The four genes are CYP1A1, CYP1B1, LINC01816, and BPIFA2. All four genes that matched the two results were upregulated genes and none of the downregulated genes matched. CYP1A1 and CYP1B1 are known to cause lung cancer by metabolizing polycyclic aromatic hydrocarbons, and long noncoding RNA is also known to play an important role in lung cancer. Considering this, we thought PM might be associated with lung cancer by activating CYP1A1, CYP1B1, and LINC01816.
颗粒物(PM)具有各种全身效应。我们使用下一代测序(NGS)研究了 PM 对肺上皮细胞的影响,并使用定量实时聚合酶链反应(qRT-PCR)对此进行了验证。我们将暴露于 PM(小于 10μm 的颗粒物)样细粉尘(ERM CZ120 细粉尘)的组培养在浓度为 50μg/mL 的一个正常肺上皮细胞系(BEAS-2B)和四个肺癌上皮细胞系(NCI-H358、HCC-827、A549、NCI-H292)中,对照组则不处理,持续培养七天。然后,我们从样本中提取 RNA 并进行 NGS。NGS 的结果显示,各种基因的表达上调或下调。在这些基因中,我们选择了平均倍数变化超过两倍且在所有五个细胞系中变化方向相同的基因。基于这些基因,我们选择了上调或下调的前 10 个基因,并用 qRT-PCR 进行验证。有四个基因与 NGS 和 qRT-PCR 结果匹配,均为上调基因。这四个基因是 CYP1A1、CYP1B1、LINC01816 和 BPIFA2。与两种结果均匹配的四个基因均为上调基因,没有一个下调基因与之匹配。CYP1A1 和 CYP1B1 通过代谢多环芳烃而导致肺癌,长非编码 RNA 也被认为在肺癌中发挥重要作用。考虑到这一点,我们认为 PM 可能通过激活 CYP1A1、CYP1B1 和 LINC01816 与肺癌有关。