通过生物信息学分析鉴定影响结直肠癌转移的新型生物标志物,并通过qRT-PCR进行验证。
Identification of novel biomarkers affecting the metastasis of colorectal cancer through bioinformatics analysis and validation through qRT-PCR.
作者信息
Lian Wenping, Jin Huifang, Cao Jingjing, Zhang Xinyu, Zhu Tao, Zhao Shuai, Wu Sujun, Zou Kailu, Zhang Xinyun, Zhang Mingliang, Zheng Xiaoyong, Peng Mengle
机构信息
Department of Clinical Laboratory, Henan No. 3 Provincial People's Hospital, Zhengzhou, 450006 Henan China.
2Department of Blood Transfusion, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052 Henan China.
出版信息
Cancer Cell Int. 2020 Mar 30;20:105. doi: 10.1186/s12935-020-01180-4. eCollection 2020.
BACKGROUND
Tumor progression and distant metastasis are the main causes of deaths in colorectal cancer (CRC) patients, and the molecular mechanisms in CRC metastasis have not been completely discovered.
METHODS
We identified differentially expressed genes (DEGs) and lncRNAs (DELs) of CRC from The Cancer Genome Atlas (TCGA) database. Then we conducted the weighted gene co-expression network analysis (WGCNA) to investigate co-expression modules related with CRC metastasis. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, DEG-DEL co-expression network and survival analyses of significant modules were also conducted. Finally, the expressions of selected biomarkers were validated in cell lines by quantitative real-time PCR (qRT-PCR).
RESULTS
2032 DEGs and 487 DELs were involved the construction of WGCNA network, and greenyellow, turquoise and brown module were identified to have more significant correlation with CRC metastasis. GO and KEGG pathway analysis of these three modules have proven that the functions of DEGs were closely involved in many important processes in cancer pathogenesis. Through the DEG-DEL co-expression network, 12 DEGs and 2 DELs were considered as hub nodes. Besides, survival analysis showed that 30 DEGs were associated with the overall survival of CRC. Then 10 candidate biomarkers were chosen for validation and the expression of CA2, CHP2, SULT1B1, MOGAT2 and C1orf115 were significantly decreased in CRC cell lines when compared to normal human colonic epithelial cells, which were consistent with the results of differential expression analysis. Especially, low expression of SULT1B1, MOGAT2 and C1orf115 were closely correlated with poorer survival of CRC.
CONCLUSION
This study identified 5 genes as new biomarkers affecting the metastasis of CRC. Besides, SULT1B1, MOGAT2 and C1orf115 might be implicated in the prognosis of CRC patients.
背景
肿瘤进展和远处转移是结直肠癌(CRC)患者死亡的主要原因,而CRC转移的分子机制尚未完全明确。
方法
我们从癌症基因组图谱(TCGA)数据库中鉴定出CRC的差异表达基因(DEGs)和长链非编码RNA(DELs)。然后进行加权基因共表达网络分析(WGCNA),以研究与CRC转移相关的共表达模块。还进行了基因本体论(GO)、京都基因与基因组百科全书(KEGG)通路分析、DEG-DEL共表达网络分析以及重要模块的生存分析。最后,通过定量实时PCR(qRT-PCR)在细胞系中验证所选生物标志物的表达。
结果
2032个DEGs和487个DELs参与了WGCNA网络的构建,并且鉴定出绿黄色、绿松石色和棕色模块与CRC转移具有更显著的相关性。对这三个模块的GO和KEGG通路分析证明,DEGs的功能密切参与癌症发病机制中的许多重要过程。通过DEG-DEL共表达网络,12个DEGs和2个DELs被视为枢纽节点。此外,生存分析表明30个DEGs与CRC的总生存期相关。然后选择10个候选生物标志物进行验证,与正常人结肠上皮细胞相比,CRC细胞系中CA2、CHP2、SULT1B1、MOGAT2和C1orf115的表达显著降低,这与差异表达分析的结果一致。特别是,SULT1B1、MOGAT2和C1orf115的低表达与CRC患者较差的生存率密切相关。
结论
本研究鉴定出5个基因作为影响CRC转移的新生物标志物。此外,SULT1B1、MOGAT2和C1orf115可能与CRC患者的预后有关。
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