| BackgroundPrimary liver cancer is a common malignant tumor of the digestive system and the fourth most common cause of cancer-related death.Hepatocellular carcinoma(HCC)is the most common pathological type of liver cancer.The characteristics of asymptomatic early stage and easy metastasis in late stage make the prognosis of HCC patients still poor though remarkable progress in the diagnosis and treatment of HCC have been made in recent decades.On the one hand,the progression of HCC is a multi-step pathological process.Under the interaction of multiple pathogenic factors,liver cells gradually develop from injury to HCC,and this process is often accompanied by liver cirrhosis.On the other hand,HCC is a highly heterogeneous tumor,and there is currently a lack of precise molecular markers for its early diagnosis.In recent years,genomic and epigenomic profiling has confirmed a series of genetic and epigenetic variations during HCC progression,which provides new opportunities to search for key molecules in HCC progression process.Therefore,based on high-throughput sequencing and microarray technology,combined with bioinformatics analysis,we tried to identify molecules that play a key role in the progression of HCC,in order to provide new methods and ideas for early diagnosis and treatment of HCC.ObjectiveThis study aims to explore biomarkers that play a key role in the progression of HCC,identify HCC targets with diagnostic,therapeutic,and predictive prognostic value,and develop diagnostic and prognostic models,which will make a significant contribution to the early diagnosis and treatment of HCC and lay a solid foundation for future functional in vivo visualization of HCC.MethodFirst,we retrieved and downloaded two datasets in the GEO database,one containing HCC and cirrhosis samples,and the other containing samples from normal liver tissue to advanced HCC during HCC progression.The two datasets were subsequently analyzed by Differentially Expressed Gene(DEG)analysis and Weighted Gene Coexpression Network Analysis(WGCNA),respectively.Based on the above analysis results,further analysis was performed in the TCGA cohort by LASSO-Cox regression and random forest algorithm to identify HCC progression molecules related to HCC prognosis,and three key HCC progression molecules were finally identified.Next,we verified the expression levels of the three molecules in cirrhotic tissue and HCC tissue by qRT-PCR,westernblot and immunohistochemistry.Moreover,the relationship between the three molecules and CIN was further investigated.Finally,a diagnostic models and a nomogram based on the three molecules were trained and validated.ResultOur study finally identified three key progression molecules,KIF20A(Kinesin Family Member 20A),TOP2A(DNA topoisomerase Ⅱ Alpha)and TTK(TTK protein kinase).In clinical samples,the expression levels of three molecules is significantly up-regulated in HCC tissues compared with adjacent cirrhotic tissues.Experiments in vitro and in vivo confirmed the ability of these three molecules to promote HCC proliferation,migration,and invasion.Furthermore,down-regulation of KIF20A,TOP2A and TTK aggravated CIN in HCC cells.Finally,the two diagnostic models constructed based on the three key molecules of HCC progression have excellent sensitivity and specificity in distinguishing HCC from cirrhotic tissue and dysplastic nodules.The nomogram constructed based on three progression key molecules combined with clinical parameters has better prognostic prediction ability than the TNM system.ConclusionKIF20A,TOP2A,and TTK are key molecules in the progression of HCC,and they are highly expressed in HCC tissue compared with normal liver tissue and liver cirrhosis tissue.Importantly,they can promote HCC proliferation,migration,invasion and inhibit HCC apoptosis,while down-regulation of KIF20A,TOP2A and TTK aggravated CIN in HCC cells.Finally,the diagnostic and predictive models constructed based on KIF20A,TOP2A,and TTK have excellent performance. |
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