Centromere protein N may be a novel malignant prognostic biomarker for hepatocellular carcinoma

Abstract

Background Hepatocellular carcinoma (HCC) is one of the deadliest tumors. The majority of HCC is detected in the late stage, and the clinical results for HCC patients are poor. There is an urgent need to discover early diagnostic biomarkers and potential therapeutic targets for HCC. Methods The GSE87630 and GSE112790 datasets from the Gene Expression Omnibus (GEO) database were downloaded to analyze the differentially expressed genes (DEGs) between HCC and normal tissues. R packages were used for Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses of the DEGs. A Search Tool for Retrieval of Interacting Genes (STRING) database was used to develop a protein-protein interaction (PPI) network, and also cytoHubba, Molecular Complex Detection (MCODE), EMBL-EBI, CCLE, Gene Expression Profiling Interactive Analysis (GEPIA), and Oncomine analyses were performed to identify hub genes. Gene expression was verified with a third GEO dataset, GSE25097. The Cancer Genome Atlas (TCGA) database was used to explore the correlations between the hub genes and clinical indexes of HCC patients. The functions of the hub genes were enriched by gene set enrichment analysis (GSEA), and the biological significance of the hub genes was explored by real-time polymerase chain reaction (qRT-PCR), western blot, immunofluorescence, CCK-8, colony formation, Transwell and flow cytometry assays with loss-of-function experiments in vitro. Results Centromere protein N (CENPN) was screened as a hub gene affecting HCC tumorigenesis. Evaluation by Cox regression showed that a high level of CENPN expression was an independent danger variable for poor prognosis of HCC. GSEA showed that high CENPN expression was linked to the following pathways: liver cancer subclass proliferation, cell cycle, p53 signaling pathway, Rb1 pathway, positive regulation of cell cycle G1/S phase transition, and DNA damage response signal transduction by p53 class moderators. Further cell experiments showed that knocking down CENPN expression decreased the proliferation and colony-forming abilities of HepG2 and Huh7 cells as well as Ki67 expression in these cell lines. The cell cycle was arrested in G1 phase, which is consistent with previous experiments on CENPN downregulation., but neither migration nor invasion were significantly affected. Western blot results revealed that the expression of p53, p27, p21, CDK4, cyclin D1, CDK2, cyclin E, pRb, E2F1 and c-myc decreased after CENPN knockdown, but there was no significant change in total Rb levels. In addition, CENPN-knockdown cells subjected to irradiation showed significantly enhanced of γ-H2AX expression and reduced colony formation. Conclusion CENPN functions as an oncogene in HCC and may be a therapeutic target and promising prognostic marker for HCC.

CENPN Acts as a Novel Biomarker that Correlates With the Malignant Phenotypes of Glioma Cells

“…First, the sample size of controls cases in both public databases and clinical cohorts from our institution was small; therefore, more studies with sufficient normal samples must be conducted in the future. Second, CENPN knockdown did not affect the cell cycle distribution, which was inconsistent with previous reports ( Oka et al, 2019 ; Wang et al, 2021 ). Thus, more glioma cell lines should be adopted for cell biological function assays.…”

“…However, studies have reported that CENPN is not indispensable in sustaining existing centromeres and recruiting kinetochore proteins because CENPN protein levels decrease before mitosis ( McClelland et al, 2007 ; Hellwig et al, 2011 ). In cancers, Wang et al (2021) and Oka et al (2019) reported that CENPN knockdown could arrest cell cycle at the G1 phase in hepatocellular carcinoma and oral squamous cell carcinoma, respectively. However, in this study, significant changes were not observed in the distribution of cell cycle phases in glioma cells subjected to CENPN knockdown.…”

“…Moreover, Sarah An et al revealed that CENPN expression was involved in the malignant progression of breast cancer patients with a history of smoking ( Andres et al, 2015 ). Most recently, CENPN was shown to promote hepatocellular carcinoma cell proliferation and CENPN deficiency was shown to increase radiotherapy-induced DNA damage ( Wang et al, 2021 ). However, the roles of CENPN in glioma have not been clarified.…”

CENPO is Associated with Immune Cell Infiltration and is a Potential Diagnostic and Prognostic Marker for Hepatocellular Carcinoma

“… 23 , 24 With advances in bioinformatics and molecular biology, an increasing number of centromere proteins have been identified as involved in the regulation of HCC. CENPA, 25 CENPE, 26 CENPF, 27 CENPH, 28 CENPK, 29 CENPL, 30 CENPM, 31 CENPN, 32 and CENPU 33 are highly expressed in HCC and their levels correlate with patient prognosis; however, to our knowledge, there have been no studies on the role of CENPO in HCC. In this study, we explored the possible mechanisms underlying the function of CENPO in HCC and its suitability as a diagnostic and prognostic biomarker.…”

A comprehensive investigation on pan-cancer impacts of constitutive centromere associated network gene family by integrating multi-omics data

“…Early studies have confirmed that CCAN was dysregulation in many kinds of tumors, then changed the process and development of tumorigenesis, finally lead to poor or good prognosis. For instance, CENPA, CENPH, and CENPN were highly upregulated in hepatocellular carcinoma, [4][5][6] colorectal cancer, [7] breast cancer, [8,9] and promoted their progression. On the contrary, reduced expression of CENPE contributes to the development of hepatocellular carcinoma.…”