Expression And Clinical Study Of Cyclophilic H In Hepatocellular Carcinoma Tissues

Background: In China,liver cancer is the fifth common malignant tumor,mainly including hepatocellular carcinoma and hepatocholangiocarcinoma and other different pathological types,of which 90% of liver cancer is hepatocellular carcinoma.Hepatocellular carcinoma(HCC),as the sixth most common malignant tumor in the world,has a five-year survival rate of only 18%.Genetics,viral hepatitis,aflatoxin exposure,smoking,obesity and diabetes are the main risk factors for hepatocellular carcinoma.Due to the insidious onset and high degree of malignancy of hepatocellular carcinoma,the existing tumor markers such as alpha-fetoprotein are difficult to diagnose early due to poor specificity,and hepatocellular carcinoma is prone to drug resistance due to poor sensitivity to radiotherapy and chemotherapy,so the current research on the treatment of hepatocellular carcinoma mainly focuses on targeted therapy and immunotherapy,so it is particularly important to find suitable targets for them.In recent years,with the development of bioinformation technology and scientific research methods,the cyclophilic family has become more and more active in the research of hepatocellular carcinoma.Cyclophilin H,also known as peptidylprolyl isomerase H(PPIH),is mainly involved in the modification of chromatin,transcription and the formation of precursor m RNA splicing complexes,recent studies have found that the high expression of PPIH in gastric malignant tumors will adversely affect the prognosis of patients,but there is still a gap in the research in hepatocellular carcinoma.Objective: The purpose of this study is to explore the expression of PPIH in hepatocellular carcinoma and its influence on the pathological characteristics of the tumor,and explore the relationship between PPIH and the prognostic factors of patients with hepatocellular carcinoma,so as to make a more accurate prediction of the prognosis of patients with hepatocellular carcinoma after surgery.In addition,by combining existing studies of the protein and functional analysis to predict its role in the development and development of hepatocellular carcinoma.Combined with the analysis of tumor immune microenvironment,exploring its value in immunotherapy for hepatocellular carcinoma is expected to make PPIH a new target for targeted therapy for hepatocellular carcinoma,which is of great significance for improving the prognosis of patients with hepatocellular carcinoma.Methods:A total of 71 paraffin samples of hepatocellular carcinoma tissues retained by the Department of Hepatobiliary Surgery of Taizhou People’s hospital after surgical resection between January 2015 and December 2019 were collected,and tumor tissues and their corresponding paracancerous tissues were stained and scored by immunohistochemistry(IHC)to evaluate the expression level of PPIH in the two tissues.The Western blot experiment was performed on three hepatocellular carcinoma and paracancerous tissue samples collected during the same period,and the differential expression of PPIH between the two groups was observed.Collect the patient’s clinical data,such as age,sex,preoperative alpha-fetoprotein(AFP)level,tumor diameter,presence or absence of vascular cancer thrombus,capsular invasion,tumor TNM stage,etc.Telephone and outpatient follow-up of patients and recording of follow-up.The scores of the patient’s immunohistochemical staining results were combined with the patient’s clinical data and matched with the follow-up results to evaluate the independent risk factors affecting the surgical prognosis of patients with hepatocellular carcinoma.The Gene Expression Profiling Data Dynamic Analysis Tool(GEPIA)was used to perform differential analysis of PPIH expression data in the Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)databases,hepatocellular carcinoma,and adjacent normal liver tissues.The Linked Omics database was used to recruit PPIH co-expressed genes and perform gene set enrichment analysis(GESA)through the Reactome,Panther,Wiki,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways.The correlation between immune cell invasion level and PPIH in hepatocellular carcinoma tissues was statistically analyzed by the Tumor IMmune Estimation Resource(TIMER).Results: 1.The expression of PPIH in tumor tissues was significantly higher than that in adjacent tissues(X~2=14.685,P<0.01).2.The expression of PPIH was positively correlated with the formation of vascular cancer thrombus(X~2=21.468,P<0.01)and tumor TNM stage(Fisher exact test =10.422,P=<0.01),and negatively correlated with the degree of tumor differentiation(Fisher precision test = 11.193,P<0.01).There were no significant implications for age(X~2=0.325,P=0.569),sex(X~2=0.037,P=0.848),tumor diameter(X~2=0.640,P=0.424),preoperative AFP level(X~2=0.403,P=0.525),and capsular invasion(X~2=0.000,P=0.989).3.Kaplan-Meier survival analysis showed that patients with high-expression PPIHexpressing hepatocellular carcinoma with low PPIH expression had a poor prognosis.4.Univariate survival analysis suggested: PPIH expression level(X~2=25.984,P<0.01),presence or absence of vascular cancer thrombus(X~2=16.762,P<0.01),TNM stage(Fisher exact test = 16.688,P<0.01)and tumor differentiation degree(Fisher precision test = 19.749,P <0.01)correlated with overall survival time.There was no correlation between age(X~2=0.174,P=0.676),sex(X~2=0.015,P=0.904),preoperative AFP level(X~2=2.214,P=0.137),tumor diameter(X~2=2.941,P=0.086),and capsular invasion(X~2=0.134,P=0.715).Multivariate analysis showed that PPIH expression level and TNM stage were independent risk factors affecting the postoperative survival of patients with hepatocellular carcinoma.5.The differential analysis of TCGA and GTEx databases suggested that the expression level of PPIH in hepatocellular carcinoma and adjacent normal tissues was statistically significant.Reactome,Panther,Wiki and KEGG pathway analysis showed that PPIH was enriched in m RNA splice,cell cycle,cytoplasmic ribosomal proteins and ribosome pathways.In addition,by studying the infiltration level of tumor immune cells of hepatocellular carcinoma in the TIMER database,it was found that the expression of PPIH was correlated with B cells(r=0.354,P<0.01),CD8+ T cells(r=0.402,P<0.01),CD4+ T cells(r=0.147,P<0.01),macrophages(r=0.395,P<0.01),neutrophils(r=0.325,P<0.01)and dendritic cells(r=0.451,P<0.01)in hepatocellular carcinoma.Studies have also found that high levels of CD4+ T cells,macrophages and neutrophils in hepatocellular carcinoma tissues indicate poor prognosis of patients.Conclusion: The expression of PPIH in hepatocellular carcinoma is significantly higher than that in adjacent tissues.The up-regulation of PPIH expression is associated with the formation of tumor vascular tumor thrombus,TNM stage and differentiation degree.The expression of PPIH has high value in the evaluation of surgical prognosis,it and tumor TNM staging are independent risk factors for poor prognosis of patients with hepatocellular carcinoma.It has been found that PPIH may be involved in m RNA splicing,cell cycle,cytoplasmic ribosomal proteins,and ribosome pathways.In addition,PPIH also played a significant role in regulating the level of immune cell invasion in hepatocellular carcinoma tissues.