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1q21.3区域驱动基因通过抑制肺腺癌细胞凋亡促进疾病进展。

The 1q21.3 region driver gene promotes disease progression via inhibition of lung adenocarcinoma cell apoptosis.

作者信息

Dong Chenchen, Li Peng, Wu Yue, Guo Zhong, He Rui

机构信息

The Second Ward of Respiratory, The Second Affiliated Hospital of Qiqihar Medical University, Qiqihar, China.

Department of Otorhinolaryngology Head and Neck Surgery, The Third Affiliated Hospital of Qiqihar Medical University, Qiqihar, China.

出版信息

Transl Cancer Res. 2022 May;11(5):1309-1320. doi: 10.21037/tcr-22-979.

DOI:10.21037/tcr-22-979

PMID:35706812

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9189215/

Abstract

BACKGROUND

Lung adenocarcinoma (LUAD) is the leading cause of cancer deaths in the world. Therefore, it is necessary to explore the underlying mechanism of (a 1q21.3 region driver gene) in the progression of LUAD cells.

METHODS

Differentially-expressed genes (DEGs) in LUAD tissues were screened based on The Cancer Genome Atlas (TCGA) database. The gene copy number variations in the 1q21.3 region were clarified by copy number variation analysis. Genes associated with overall survival (OS) were identified by Kaplan-Meier (KM) analysis. The intersection of the genes was used to obtain the driver genes. LUAD patients were grouped with driver gene expression, and the DEGs were identified. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was used to identify the enrichment pathways of the driver genes. was knocked down using lentiviral infection in A549 and PC9 cell lines. The efficiencies of lentiviral infection were confirmed by RT-PCR (reverse transcription polymerase chain reaction). After knockdown, changes in cell viability were confirmed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, changes in cell proliferation and apoptosis were confirmed by enzyme-linked immunosorbent assay (ELISA), while changes in the expression of apoptosis-related proteins ( and ) were confirmed by RT-PCR and western blot.

RESULTS

A total of 483 LUAD samples and 59 normal control samples were obtained from TCGA database, and 640 upregulated genes were identified. 154 genes with a coefficient of copy number variations greater than 10% in the 1q23.1 region and 1,247 genes that were significantly associated with patient OS were selected. The intersection results indicated that EFNA3 might be a driver gene of LUAD. KEGG enrichment analysis indicated that the DEGs were mainly enriched in apoptosis-related pathways. Cell experiments showed that after lentiviral knockdown of , messenger RNA (mRNA) and protein expression was significantly reduced (P<0.05), cell viability was markedly reduced (P<0.05), LUAD cell apoptosis increased notably (P<0.05), and LUAD cell proliferation decreased significantly (P<0.05). After knockdown, the expression of apoptosis-related proteins ( and ) and mRNA in LUAD cells increased markedly (P<0.05).

CONCLUSIONS

As a driver gene in the progression of LUAD, mainly affects the progression of LUAD by regulating LUAD cell apoptosis.

摘要

背景

肺腺癌（LUAD）是全球癌症死亡的主要原因。因此，有必要探索（一个1q21.3区域驱动基因）在LUAD细胞进展中的潜在机制。

方法

基于癌症基因组图谱（TCGA）数据库筛选LUAD组织中的差异表达基因（DEG）。通过拷贝数变异分析阐明1q21.3区域的基因拷贝数变异。通过Kaplan-Meier（KM）分析确定与总生存期（OS）相关的基因。利用基因的交集获得驱动基因。根据驱动基因表达对LUAD患者进行分组，并鉴定DEG。使用京都基因与基因组百科全书（KEGG）富集分析来确定驱动基因的富集途径。在A549和PC9细胞系中通过慢病毒感染敲低（基因名称未给出）。通过逆转录聚合酶链反应（RT-PCR）确认慢病毒感染效率。敲低后，通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法确认细胞活力变化，通过酶联免疫吸附测定（ELISA）确认细胞增殖和凋亡变化，同时通过RT-PCR和蛋白质印迹确认凋亡相关蛋白（和基因名称未给出）的表达变化。

结果

从TCGA数据库中获取了总共483个LUAD样本和59个正常对照样本，并鉴定出640个上调基因。选择了1q23.1区域中拷贝数变异系数大于10%的154个基因和与患者OS显著相关的1247个基因。交集结果表明EFNA3可能是LUAD的驱动基因。KEGG富集分析表明DEG主要富集在凋亡相关途径中。细胞实验表明，慢病毒敲低（基因名称未给出）后，（基因名称未给出）信使核糖核酸（mRNA）和蛋白质表达显著降低（P<0.05），细胞活力明显降低（P<0.05），LUAD细胞凋亡显著增加（P<0.05），LUAD细胞增殖显著降低（P<0.05）。敲低（基因名称未给出）后，LUAD细胞中凋亡相关蛋白（和基因名称未给出）和mRNA的表达明显增加（P<0.05）。