The Establishment Of Scoring Model For Genes Associated With Liver Metastasis In Pancreatic Cancer And The Molecular Mechanism Of The Screened Promoting Metastatic Gene KPNA4

Background and purpose: Pancreatic cancer is one of the most lethal malignancies worldwide,and is very easy to metastasize to liver.Meanwhile,a shorter overall survival was observed in pancreatic cancer patients with isolated hepatic metastases,compared to patients with metastasis to other sites.However,the mechanisms underlying the metastasis to liver in pancreatic cancer have not yet been well explained.Methods: The differential gene expression analysis was used to identify the liver metastasis-related genes.The single-sample gene set enrichment analysis was employed to construct a scoring system Liver Metastasis Score(LMS).The correlation analysis and survival analysis were adopted to evaluate the predictive value of LMS.In addition,to confirm the impact of KPNA4 on the functionalities of pancreatic cancer cells,the present study selected two widely used pancreatic cancer cell lines,and silence the KPNA4 gene by si RNAs within these cells.The impact of KPNA4 silence on cell proliferation,invasion,and migration was evaluated by CCK-8,flow-cytometry.Combined with the enrichment results of high-throughput information,Western Blot was further employed to investigate the potential mechanism of KPNA4 promoting the metastasis potential of pancreatic cancer cells.Results: In this study,we identified a total of 154 genes upregulated in primary tissues of pancreatic cancer with liver metastasis using the Genome Cancer Atlas(TCGA)and GSE151580 cohorts,which were defined a liver metastasis score(LMS)to assess the risk of liver metastasis in pancreatic cancer.Further gene enrichment analysis indicated that LMS related genes were mainly enriched in the functional items related to epithelial-mesenchymal transformation,estrogen response,p53 and glycolysis pathway.We found LMS related genes were closely associated with disease type and tumor grade of pancreatic cancer,and its prognostic value has been validated in three cohorts with long-term follow-up(TCGA,GSE71729,and E-MTAB-6134).Furthermore,liver metastasis-related genes were primarily found in malignant ductal cells by integrative analysis of the bulk and single-cell gene expression data,and malignant ductal cells and M0 macrophages were highly correlated with LMS,indicating that the two cell types might function as tumor-promoting cells in pancreatic cancer.To further uncover the critical regulator in liver metastasis of pancreatic cancer,the integrative analysis revealed that KPNA4 achieved the highest specificity in malignant ductal cells,and exhibited extremely high expression in pancreatic cancer with liver metastasis.We further confirmed that that KPNA4 silence could efficiently suppress the proliferation,invasion,and migratory abilities of pancreatic cancer cells through p53 signal axis,suggesting that KPNA4 may be closely associated with liver metastasis in pancreatic cancer.Conclusion: In summary,we constructed a LMS score to evaluate the risk of liver metastasis in pancreatic cancer by systematic analysis.KPNA4 could promote the proliferation,invasion,and migratory abilities of pancreatic cancer cells through p53 pathway.