Study On The Relationship Between Prr11 And Circulating Immune Cells And Tumor Metastasis And Recurrence In Hepatoma Inflammation Microenvironment

Objective:To analyze the expression of proline-rich protein 11(PRR11)in the human hepatocellular carcinoma(HCC)and investigate the effects of RNA interference targeted silence PRR11 gene on the proliferation and migration and invasion of HCC cell line SMMC-7721.and compare and validate the prognostic abilities of preoperative systemic immune cells in hepatocellular carcinoma(HCC)after curative hepatectomy.Methods:Immunohistochemistry and western blotting were used to analyze the expression of PRR11 protein in HCC tissues and its cancer-adjacent tissues,and the relationship between PRR11 and clinical pathological parameters was studied.Western blotting was also used to analyze the expression of PRR11 protein in HCC cell lines SMMC-7721,HepG2 and normal liver cell L02.RNA interference targeted silence PRR11 gene was transfected into SMMC-7721 cells,The proliferation and migration and invasion of the transfected cells were assessed by CCK-8 assay and wound healing and Transwell.We developed two nomograms to predict the postoperative recurrence-free survival(RFS)and overall survival(OS)after comparisons of the systemic immune cell prognostic scores.The two nomograms were constructed based on 305 patients who underwent curative hepatectomy for HCC.The predictive accuracy and discriminative ability of the nomograms were compared with six commonly used staging systems for HCC.Results:The results of immunohistochemistry and western blotting demonstrated that the expression of PRR11 in HCC tissues was found to be significantly higher than that of cancer-adjacent tissues(1.114±0.215 and 0.644±0.071),and the expression of PRR11 protein was correlated with tumor size,portal vein invasion,TMN stages,Edmondson grade,metastasis and NMLR(P<0.05).The relative amount of PRR11 protein in SMMC-7721 and HepG2 was significantly higher than that in L02(1.219±0.034?1.213±0.081 and 0.839±0.038).It was showed that downregulation of PRR11 inhibited cell proliferation and migration and invasion(P<0.05).The results were vali-dated using bootstrap resampling and an internal validation cohort of 142 patients and an external validation cohort of 169 patients.Necroinflammatory activity in peritumoral liver tissues in the primary cohort was evaluated by hematoxylin and eosin(H&E)staining.Neutrophil,monocyte,and lymphocyte ratio(NMLR)had a higher area under the receiver operating characteristic curves(AUROC)value at both RFS(AUC = 0.603)and OS(AUC = 0.726)compared to that of other systemic immune cell prognostic scores.The independent predictors of RFS or OS,including a-fetoprotein(AFP),tumor size,tumor number,mi-crovascular invasion,and NMLR,were incorporated into the two nomograms.In the primary cohort,the C-indexes of the RFS and OS nomograms were 0.705 and 0.797,respectively.The ROC analyses showed that the two nomograms had larger AUC values(0.664 for RFS and 0.821 for OS)than those of the American Joint Commission on Cancer seventh edition,Barcelona Clinic Liver Cancer,Cancer of the Liver Italian Program,Japan Integrated Staging Score,Okuda stage,and the Vauthey's system.These results were verified by internal and external validations.The nomogram-predicted probability of RFS was as-sociated with peritumoral necroinflammatory activity scores(r=0.304,P<0.001).Conclusion:The expression of PRR11 protein in HCC tissues was significantly higher than that in cancer-adjacent tissues,and RNAi of PRR11 in the SMMC-7721 cells may inhibit cell proliferation,migration and invasion.The proposed nomograms containing independent predictors of NMLR had accurate prognostic prediction in patients with HCC after curative hepatectomy.