Construction Of A Multigene Survival Prediction Model For Hepatocellular Carcinoma And Study On Expression And Biological Function Of KIF20A In Hepatocellular Carcinoma

Objectives: About 75% of PLCs are HCC,and the prognoses of HCC patients are poor.The development of new survival prediction models can help physicians make prognostic assessments and treatment decisions for HCC patients.Previous studies have shown that KIF20 A is highly expressed in various malignant tumors and plays an important role in promoting the malignant biological behaviors of cancers.Nevertheless,its role in HCC is unclear and remains to be further investigated.Materials and Methods: Based on data of liver cancer patients from TCGA,Cox regression and LASSO regression were utilized to establish a new polygenic risk score.Univariate and multivariate Cox regression analysis and the Kaplan-Meier method were used to evaluate the prognostic value of risk scores and prognosis-related differentially expressed genes.Enrichment analysis of DEGs between patients in the high and low risk groups was conducted by using GO and KEGG.ss GSEA was used to assess differences in immune cells and function between patients in the high and low risk groups.q RT-PCR was used to assess the expression levels of genes in clinical samples.The immunohistochemical staining results in the HPA database showed the protein expression levels of genes in liver cancer tissues and normal tissues.A nomogram was constructed based on the polygenic risk score and patient clinical indicators,and the model accuracy and clinical application value were evaluated through calibration curves,decision curves and ROC curves.Data from TCGA and GTEx were used to analyze KIF20 A expression in various tumor and normal tissues.In the second part,the protein expression of KIF20 A in 48 pairs of HCC tissues and adjacent normal tissues were analyzed by immunohistochemical staining.The protein expression of KIF20 A in 12 pairs of HCC tissues and adjacent normal tissues were analyzed by WB.The m RNA expression of KIF20 A in 20 pairs of HCC tissues and adjacent tissues were analyzed by q PCR.The expression levels of KIF20 A in normal liver cell and liver cancer cells were analyzed by WB and q PCR experiments.In addition,we explored the effect of KIF20 A on the malignant biological behavior of HCC cells by CCK-8 assay,wound healing assay and transwell invasion assay.The underlying mechanism of KIF20 A in facilitating migration and invasion related to EMT was investigated by the western blot.Results: Finally,we developed a novel five-gene(KIF20A,CENPA,HMMR,G6 PD,ADH4)risk score.Based on the median value of risk score,patients were separated into high-risk and low-risk groups.The OS of patients in the high-risk group was significantly worse than that in the low-risk group.And this risk score was an independent risk factor associated with OS of HCC patients.In addition,we developed a prognostic nomogram consisting of a polygenic risk score and TNM stage.Decision curve,calibration curve and ROC curve results showed that polygenic risk score and nomogram have good predictive ability and clinical application value.In addition,the m RNA expression levels of ADH4 were relatively low in HCC tissues,whereas the m RNA expression levels of KIF20 A,CENPA,HMMR,and G6 PD were relatively high in HCC tissues.The results of immunohistochemistry,q PCR and WB experiments showed that KIF20 A was highly expressed in HCC tissues and HCC cells.Analysis of cell function experiments found that KIF20 A played an important role in promoting growth,invasion and migration of HCC cells.Furthermore,KIF20 A promotes HCC migration and invasion by regulating the EMT process.Conclusions: We have developed a novel 5-gene risk score that can predict the prognosis of HCC patients.These five genes(KIF20A,CENPA,HMMR,ADH4,and G6PD)may be potential therapeutic targets and biomarkers for HCC.For the first time,we have developed a computational procedure for our five-gene risk score and a dynamic nomogram,which may be useful risk stratification and survival prediction tools for HCC patients.High expression of KIF20 A suggests a poor prognosis for HCC patients.KIF20 A may promote cell migration and invasion of HCC by regulating the EMT process.And it could be an effective therapeutic target for HCC patients.