Correlation Between Vasculogenic Mimicry And Clinical Prognosis In Hepatocellular Carcinoma

Objective:Our main purpose was to detect the expression of vasculogenic mimicry and angiogenic molecules VEGF,epithelialmesenchymal transition related molecules such as Twist-1,E-cadherin,Vimentin and EphA2,as well as VE-cadherin contributed to vasculogenic mimicry process in hepatocellular carcinoma.Then,to figure out the correlation between expression of selected molecules and clinicopathologic data.The main factors affecting clinical prognosis were screened out to provide new ideas for prognosis evaluation and treatment of patients with hepatocellular carcinoma.Methods:Clinicopathologic and follow-up data of 132 patients with a diagnosis of hepatocellular carcinoma from Hunan Provincial People’s Hospital between October 2012 and December 2013 were collected for retrospective analysis.Firstly.The experimental group of hepatocellular carcinoma tissue and the control group of carcinomatous adjacent tissue were sectioned consecutively,the first slide was stained with CD34 by immunohistochemical staining and stained with PAS to observe the presence of VM in the experimental group.Second,serial section of the experimental group and the control group were stained with VE-Cadherin,VEGF,EphA2,Twist1,E-cadherin and Vimentin by immunohistochemical staining.Thirdly,statistical analysis was made on the difference between the above molecular expression in the experimental group and the control group,as well as the correlation between each molecule and VM,so as to further analyze the relationship between VM and the patients’ gender,age,tumor size,TNM stage,Edmondson stage,portal cancerous thrombus and liver cirrhosis.Fourthly,the correlation between clinicopathologic data and the molecular expression was analyzed to screen out prognostic risk factors.Finally,TCGA database was used to verify the relationship between VM related molecules screened in this study and the prognosis of hepatocellular carcinoma on the level of mRNA.Results :(1)VM was found in 44(44/132,33.3%)hepatocellular carcinoma tissue by stained with CD34 and PAS.(2)The expression of VE-cadherin,VEGF,EphA2,Twist1 in hepatocellular carcinoma tissue were all higher than those in carcinomatous adjacent tissue(positive rates: 68.2%,51.5%,56.8%,57.6% in the experimental group,and 19.7%,11.4%,25%,21.2% in the control group).The expression of E-cadherin in hepatocellular carcinoma tissue were lower than those in carcinomatous adjacent tissue(positive rates:48.5% in the experimental group,and 65.9% in the control group).The expression of Vimentin in tumor cells and normal hepatocytes were negative.(3)For the VM with clinicopathologic data(gender,age,tumor size,TNM stage,Edmondson stage,portal cancerous thrombus and liver cirrhosis)were compared and analyzed,and finds out VM positive rate of patients with III+IV stage and portal cancerous thrombus were higher than patients with I+II stage and without portal cancerous thrombus.(4)Expressions of Twist1 and VEGF in hepatocellular carcinoma were positively correlated with the presence of VM.(5)The survival time of patients with different TNM stages,Edmondson grades,with or without portal cancerous thrombus and VM,and whether or not Twist1 and VEGF were expressed was different.Further multi-factor analysis showed that VM,Twist1 and Edmondson grades were independent risk factors affecting the survival time of hepatocellular carcinoma.(6)The relationship between the screened VM related molecules and the prognosis of hepatocellular carcinoma in this study is consistent with the results of TCGA database analysis.Conclusion:(1)VM、VE-Cadherin、VEGF、EphA2、Twist1、E-cadherin and Vimentin may be involved in the occurrence and development of hepatocellular carcinoma.(2)VM has a high incidence in patients with advanced hepatocellular carcinoma,which may be contribute to hematological metastasis and promote the formation of portal cancerous thrombus.(3)VEGF and Twist1 are closely related to the formation of VM.(4)TNM stage,Edmondson grades,portal cancerous thrombus,Twist1,VEGF and VM were the factors of patient overall survival time.The presence of VM,upregulation of Twist1 and Edmondson III-IV grade were independent risk factors for poor prognosis in patients with hepatocellular carcinoma.