The Role Of Prrx1 Mensenchymal Circulating Tumor Cells In The Rugulation Of Recurrence And Its Mechanism In Hepatocellular Carcinoma

Low expression of Prrx1 mesenchymal CTCs in peripheral blood of HCC patients promote tumor recurrenceObjective To analyze the correlation between clinical characteristics of HCC patients and epithelial-mesenchymal phenotype circulating tumor cells in peripheral blood.Methods 77 HCC patients and 21 benign liver disease patients at Zhujiang Hospital of Southern Medical University from April 2015 to December 2016 were retrospective observed in this study.We adopted the CanPatrolTM System to detect epithelial-mesenchymal phenotype CTCs in peripheral blood.To clarify the epithelial-mesenchymal phenotype CTCs specificity originated from the tumor and investigate the correlation between CTCs and clinical features.Results According to the epithelial and mesenchymal markers,CTCs were divided into three subtypes:epithelial,mesenchymal and mixed CTCs.Epithelial-mesenchymal phenotype CTCs were detected in none of benign liver disease patients,but the positivity rate of epithelial,mixed and mesenchymal CTCs in peripheral blood of 77 HCC patients were 37.7%(29/77),74.0%(57/77)and 50.6%(39/77)respectively.And the positivity rate of epithelial-mesenchymal phenotype CTCs in HCC patients were significantly higher than the benign liver disease group(P<0.05).In our study,the positivity rate of epithelial CTCs was significantly correlated with AFP,number of tumor and portal vein tumor thrombus(P<0.05),mixed CTCs was significantly correlated with portal vein tumor thrombus,tumor differentiation and BCLC stage(P<0.05),and mesenchymal CTCs was significantly correlated with tumor size、portal vein tumor thrombus,tumor differentiation and BCLC stage(P<0.05).Further analysis found that the recurrence rate(83%,5/6)of the Prrx1 low expression group was higher than that of the high expression group(10%,1/10)(P<0.05)Conclusions epithelial,mixed and mesenchymal CTCs can reflect the clinical characteristics of HCC patients in different aspects,which may serve as a indicator for evaluating the progression of HCC.Meanwhile,low expression of Prrx1 mesenchymal CTCs can be used as biomarkers to monitor the recurrence of HCC.Effect of Prrxl on biological characteristics of hepatocellular carcinoma cellsObjective To investigate the effect of Prrx1 expression on the phenotype,invasiveness and stem cell characteristics of hepatoma cells.Methods Firstly,the differences of Prrx1 protein and mRNA expression in five different hepatocellular carcinoma cell lines were compared by Western blot and qPCR.The hepatocellular carcinoma cells with different Prrx1 expression levels were selected by further interfering experiments.Further observation of cell morphological changes in parallel cell function experiments,the Prrx1 differential expression on the biological characteristics of hepatocellular carcinomaResults We found that in five hepatocellular carcinoma cells,the highest expression level of Prrx1 was found in hepatocellular carcinoma BEL-7404.The BEL-7404 cells with low Prrx1 expression were obtained by lentivirus infection.The down-regulation of Prrx1 induces the mesenchymal phenotype of BEL-7404 induced by EMT,and the number of cells invaded by BEL-7404 cells with Prrx1 expression is significantly increased compared with the negative control group(P<0.05).We further detected the stemness indicators(CD 133,Nanog and Sox2)by Western blot and qPCR in two groups of cells,and found that the expression of stemness was up-regulated in BEL-7404 cells with down-regulated Prrx1 expressionConclusions Down-regulation of Prrx1 induces mesenchymal phenotype of EMT in hepatocarcinoma cells and induces its invasive ability and tumor stem cell characteristicsTo analyze the correlation between clinical characteristics of HCC patients and epithelial-mesenchymal phenotype circulating tumor cells in peripheral blood.And We found that mensenchymal CTCs and hepatocellular carcinoma metastasis are positively correlated,and low expression of Prrx1 mensenchymal CTCs are more likely to form metastases.In addition,according to our in vitro experiment:Prrx1 in Bel-7404 cells down-regulation can induce its occurrence epithelial-mesenchymal transition(EMT)and contribute to its invasion and obtain the characteristics of tumor stem cells.Therefore,We hypothesized that the low expression of Prrx1 mesenchymal CTCs could enhance the invasion ability and obtain the characteristics of tumor stem cells.On the basis of 2 leves cells-clinical,this project reveals that Prrx1 induced hepatocellular carcinoma mesenchymal CTCs to form metastases,clarifies the molecular mechanism of Prrx1 in the process of CTCs target metastasis and provides scientific basis for precise treatment.