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肝癌细胞中 DAPK1 敲除后的差异基因表达分析。

Differential gene expression analysis after DAPK1 knockout in hepatocellular carcinoma cells.

机构信息

Guangdong Medical University, Zhanjiang, China.

出版信息

PeerJ. 2022 Aug 2;10:e13711. doi: 10.7717/peerj.13711. eCollection 2022.

Abstract

BACKGROUND

The mechanism through which death-associated protein kinase 1 (DAPK1) causes hepatocellular carcinoma (HCC) progression remains unclear. In this study, we aimed to identify key proteins that were altered after DAPK1 knockout.

METHODS

Stable DAPK1 knockout HCC cell lines were established, then the differentially expressed genes (DEGs) of HCC were screened using the NetworkAnalyst database and enriched using the Metascape software. Protein-protein interaction networks (PPIs) were analyzed and visualized using the STRING database expansion.

RESULTS

In total, 732 differentially expressed genes were identified, including 415 upregulated genes and 317 downregulated genes. Through Cytoscape software scoring, 10 pivotal genes were found to be closely related to changes in DAPK1 expression; Kininogen-1 (KNG1), Complement C3 (C3), Metalloproteinase inhibitor 1 (TIMP1), and Alpha-2-HS-glycoprotein (AHSG) were the most strongly associated with DAPK1 expression changes. Moreover, western blot analysis results revealed that changes in the levels of proteins encoded by the four key genes after DAPK1 knockout were consistent with those seen in the database screening.

CONCLUSIONS

These results provide a direction for further studies on the DAPK1 gene and on the mechanism through which DAPK1 leads to hepatocellular carcinoma development.

摘要

背景

凋亡相关蛋白激酶 1(DAPK1)导致肝细胞癌(HCC)进展的机制尚不清楚。在这项研究中,我们旨在鉴定 DAPK1 敲除后发生改变的关键蛋白。

方法

建立稳定的 DAPK1 敲除 HCC 细胞系,然后使用 NetworkAnalyst 数据库筛选 HCC 的差异表达基因(DEGs),并使用 Metascape 软件进行富集分析。使用 STRING 数据库扩展分析和可视化蛋白质-蛋白质相互作用网络(PPIs)。

结果

共鉴定出 732 个差异表达基因,包括 415 个上调基因和 317 个下调基因。通过 Cytoscape 软件评分,发现 10 个关键基因与 DAPK1 表达变化密切相关;激肽原-1(KNG1)、补体 C3(C3)、金属蛋白酶抑制剂 1(TIMP1)和α-2-HS-糖蛋白(AHSG)与 DAPK1 表达变化的相关性最强。此外,Western blot 分析结果显示,DAPK1 敲除后四个关键基因编码蛋白水平的变化与数据库筛选结果一致。

结论

这些结果为进一步研究 DAPK1 基因及其导致肝细胞癌发展的机制提供了方向。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0675/9354754/40e3c0da7db4/peerj-10-13711-g001.jpg

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