Department of Thoracic Surgery, Shenzhen People's Hospital, The Second Clinical Medical College of Jinan University.
J Clin Lab Anal. 2022 Sep;36(9):e24628. doi: 10.1002/jcla.24628. Epub 2022 Aug 5.
Lung cancer ranking high in the cancer-related list has long perplexed patients, in which glucosamine-phosphate N-acetyltransferase 1 (GNPNAT1) is found to be highly expressed. Besides, DNA methylation is perceived as a biomarker to assess the prognosis of patients with various cancers. However, the correlation between GNPNAT1 and DNA methylation and the role of GNPNAT1 in lung cancer remain vague.
Principal component analysis (PCA), heatmap, volcano map, Venn diagram, gene ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were used to screen out the candidate genes. The viability, migration, and invasion of lung cancer cells were detected by CCK-8 and Transwell assays. An xenograft tumor mouse model was established. The relative expressions of GNPNAT1, E-cadherin, vimentin, Matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-2 (TIMP-2), E2F1, and cyclin D1 in cells or xenograft tumor tissues were quantified by Western blot, RT-qPCR, or immunohistochemistry assay.
GNPNAT1 was screened as the research object. GNPNAT1 methylation was downregulated, while GNPNAT1 expression was upregulated in lung cancer tissues. The methylation and mRNA levels of GNPNAT1 were correlated with the patient prognosis. GNPNAT1 increased cell viability, migration and invasion, and promoted the xenograft tumor volume and weight, whereas shGNPNAT1 acted oppositely. Moreover, expressions of Vimentin, MMP-2, E2F1, and cyclin D1 were increased, but E-cadherin and TIMP-2 expressions were decreased by overexpressed GNPNAT1, whilst GNPNAT1 knockdown ran conversely.
GNPNAT1 and methylated GNPNAT1 coverage are biomarkers for the diagnosis and prognosis of lung cancer.
在癌症相关列表中排名较高的肺癌一直困扰着患者，在该癌症中发现葡萄糖胺-磷酸 N-乙酰转移酶 1（GNPNAT1）高度表达。此外，DNA 甲基化被认为是评估各种癌症患者预后的生物标志物。然而，GNPNAT1 与 DNA 甲基化之间的相关性以及 GNPNAT1 在肺癌中的作用仍不清楚。
采用主成分分析（PCA）、热图、火山图、韦恩图、基因本体（GO）和京都基因与基因组百科全书（KEGG）分析筛选候选基因。通过 CCK-8 和 Transwell 测定检测肺癌细胞的活力、迁移和侵袭。建立异种移植肿瘤小鼠模型。通过 Western blot、RT-qPCR 或免疫组织化学检测细胞或异种移植肿瘤组织中 GNPNAT1、E-钙黏蛋白、波形蛋白、基质金属蛋白酶-2（MMP-2）、金属蛋白酶组织抑制剂-2（TIMP-2）、E2F1 和细胞周期蛋白 D1 的相对表达。
筛选出 GNPNAT1 作为研究对象。GNPNAT1 甲基化下调，而肺癌组织中 GNPNAT1 表达上调。GNPNAT1 的甲基化和 mRNA 水平与患者预后相关。GNPNAT1 增加细胞活力、迁移和侵袭，并促进异种移植肿瘤体积和重量，而 shGNPNAT1 则作用相反。此外，过表达 GNPNAT1 增加了波形蛋白、MMP-2、E2F1 和细胞周期蛋白 D1 的表达，而降低了 E-钙黏蛋白和 TIMP-2 的表达，而 GNPNAT1 敲低则相反。
GNPNAT1 和甲基化 GNPNAT1 覆盖率是肺癌诊断和预后的生物标志物。
