| Background:Primary carcinoma of the liver is a typical inflammation-related tumor,which is often secondary to chronic hepatitis and cirrhosis.A large number of immune cells are found in the tumor microenvironment.Regulatory T cells(Treg),an important phenotype of CD4+T cells,plays an immunoregulatory role.Recent studies have shown that regulatory T cell(Treg)could promote the development and progression of primary carcinoma of the liver by inhibiting the killing effect of effector cells such as CD4+T,CD8+T and natural killer cells.CD39 and CD73,two adenosine metabolism molecules,are expressed on Treg membrane,which synergistically hydrolyze ATP to produce adenosine,thereby exerting immunosuppressive effects.In this paper,the research was conducted to mainly study the proportion change and the clinical significance of Treg,CD39 and CD73 subgroups in peripheral blood in patients with primary liver cancerObjectives:To analyze the relationship between the clinical progress of hepatocellular carcinoma and the change of Treg,CD39 and CD73 positive subgroups via detecting the proportion of these cells in peripheral blood of patients with the primary carcinoma,and at the same time,to observe the correlation between the proportion of Treg,CD39 and CD73 positive subgroups and the indicators of liver injury in patients with the carcinoma.And further to provide new ideas for the diagnosis and treatment of primary liver cancer.Methods:1.The peripheral blood samples were collected from 80 patient with primary liver cancer diagnosed according to clinical or/and pathological standard.These patients were as the study group in strict accordance with the Inclusion and Exclusion Criteria,who were admitted to the Hepatobiliary and Pancreatic Center of Jiangsu Subei People's Hospital from December 2017 to March 2019.Based on the primary liver cancer staging,the patients in this study group were categorized into Stage ?-?(36 cases)and Stage ?-?(44 cases).Meanwhile,the indicators of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in these patients were measured,and the patients of 51 were divided into liver injury group and the patients of 29 into without liver injury group according to the standard(ALT?80IU/L)of the ALT two times the normal value.At the same time,the "healthy" volunteers were selected as the control group and their peripheral blood samples were collected2.Flow cytometry was used to detect the proportion of Treg and its CD39 and CD73 positive subgroups in peripheral blood from the both groups respectively.The proportion differences of Treg and its CD39 and CD73 positive subgroups in the blood were compared between the study and the control group,and furtherly,this proportion differences were compared between patients in stage ?-? and stage ?-?in the study group.The comparisons of the proportion differences of Treg and its CD39 and CD73 positive subgroups also were performed between the liver injury and without liver injury group3.The corelationship between the proportion of CD39 and CD73 positive subgroups of Treg for patients with the carcinoma and the liver function damage indicators of ALT and AST were analyzedResults:1.There were no statistically significant differences in gender composition,age,etiology and family history between the study group and the "healthy" volunteer control,between the stage ?-? and the stage ?-? group,and between the liver injury and the non-liver injury group(P>0.05).2.Comparison of Treg and its CD39 and CD73 positive subgroups between the study and the control group:The proportion of Treg in CD4+T cells in peripheral blood of primary liver cancer patients(study group)was significantly higher than that in the control group(9.57±3.40)VS(5.73±2.03),and the difference was statistically significant(P<0.05).Meanwhile,the proportion of CD39+Treg(18.02±9.89)and CD73+Treg(15.59±5.87)in the blood of the study group was significantly higher than that of the control group(13.49±2.16 and 13.46±6.67),and the difference was statistically significant(P<0.05).Although the proportion of CD39+CD73+Treg in the blood of the study group was higher than that of the control group(4.95±1.17)VS(4.88±1.23),there was no statistical difference between the groups(P>0.05).3.Comparison of Treg and CD39 and CD73 positive subgroups in the different liver cancer staging groups:The proportion of Treg in CD4+T in the blood of stage ?-? patients was significantly higher than that of stage ?-? patients(10.52±3.42)VS(8.40±3.03),and the difference was statistically significant(P<0.05).Analysis of CD39+Treg also found that the proportion of Treg in stage ?-? patients was significantly higher than that in stage ?-? patients(20.01±9.84)VS(15.57±9.51),and the difference was statistically significant(P<0.05).However,the CD73+Treg study found no statistical difference in the proportion between the both groups(P>0.05),and the results were 15.45±5.99 and 15.76 ± 5.80,respectively.Further the comparison of CD39+CD73+Treg analysis also found similar results with CD73+Treg,with values of 5.01±1.08 and 4.89±1.24 respectively(P>0.05).4.Comparison of Treg and its CD39 and CD73 positive subgroups in the liver injury and the non-liver injury group:The proportion of Treg in CD4+T cells in peripheral blood of patients in the liver injury group was significantly higher than that in the non-liver injury group:(10.27±3.41)VS(8.31±3.06),the difference was statistically significant(P<0.05).Meanwhile,the proportion of CD39+Treg(19.86±10.58)and CD73+Treg(16.61±6.15)in the blood of the liver injury group was also higher than that of the non-liver injury group(14.77±7.67 and 13.73±5.07),and the difference was statistically significant after statistical analysis(P<0.05).Moreover,the analysis of the proportion of CD39+CD73+Treg in Treg showed that although the liver injury group(5.09±1.09)was higher than the non-liver injury group(4.68±1.27),there was no statistically significant difference between the both groups(P>0.05)Correlation analysis was conducted on the proportions of ALT or AST and Treg respectively,and it was found that there were positive correlations,with correlation coefficient(r)of 0.578 and 0.581 respectively,P<0.05.The analysis of ALT and CD39+Treg proportion also found a positive correlation(r=0.556,P<0.05),and the AST and CD39+Treg proportion was also positive correlation,with a correlation coefficient of 0.609,P<0.05.Although the ALT or AST and CD73+Treg proportion are positively correlated,their correlations are relatively weak compared with the above correlation coefficients,and their values are 0.279 and 0.342 respectively,P<0.05Conclusions:1.According to the proportion of Treg in CD4+T cells in peripheral blood of patients in the study group was higher than that in the control group,it can be concluded that this cells play an important role in the occurrence and development of primary hepatocellular carcinoma2.From comprehensively analyzing the difference of CD39+Treg and CD73+Treg ratio between the study and the control group,and the difference in different stages of the tumors it is reasonable to speculate that CD39 and CD73 can promote Treg to exert immunosuppressive effect in the immune microenvironment of primary hepatocellular carcinoma,but CD39 may have stronger inhibitory effect3.By analyzing the results from the proportion of Treg,CD39+Treg and CD73+Treg between the non-hepatic injury and the combined hepatic injury group,and the positive correlation between the cell proportion and the indicators of ALT or AST for primary hepatocellular carcinoma,it can be concluded that Treg,CD39 and CD73 positive subgroups may promote the invasion and compression of the tumor to causing liver injury. |
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