| Objective This study intends to observe effects of radiotherapy on the number of CD4+CD25+CD127-regulatory T cells(Treg),CD3+CD8+effector T cells(Teff,namely CTL)and the expression of related cytokines(i.e.IL-2、IL-10、IL-6,TGF-β,HMGB1 and PGE2)in the blood of patients with esophageal squamous cell carcinoma.To analyze the correlation of those immune cells and cytokines with the short-term efficacy of patients with esophageal squamous cell carcinoma,and to provide experimental basis and theoretical basis for the clinical use of immunotherapy combined with radiotherapy for esophageal cancer.Methods A total of 56 patients with esophageal squamous cell carcinoma admitted to the Oncology Department of Taizhou People’s Hospital from October 2019 to October 2020 were selected as the esophageal cancer research group,all patients were planned to undergo radical radiotherapy.Ten healthy people who underwent physical examination in our hospital during the same period were selected as the control group.To collect venous blood 5ml in the morning from patients with esophageal squamous cell carcinoma within 1 week before radiotherapy,about 3 weeks after the start of radiotherapy,and within 1 week after radiotherapy.Blood samples were divided into two parts.One part was used for flow cytometry to detect the number of CD4+CD25+CD127-Treg and CD3+CD8+Teff in peripheral blood.Cell Quest software was used for analysis,and the proportion of Treg cells in CD4+T lymphocytes and Teff cells in peripheral blood lymphocytes were respectively used to reflect the number of Treg and Teff.The other part of whole blood was centrifuged to collect serum and stored in a refrigerator at-80℃.The concentrations of IL-10,IL-2,IL-6,TGF-β,HMGB1 and PGE2 in serum were detected by ELISA.The short-term efficacy of radiotherapy was evaluated 1 to 3 months after radiotherapy.SPSS26.0 software was used for statistical analysis.After normal test,the data conforming to normal distribution was represented by X±S,and the non-normal distribution data was represented by M(P25,P75).The statistical data were compared byχ2 test,and the mean of two samples was compared by two independent samples t test or rank sum test.One-way repeated measure ANOVA was used for pairwise comparison of multiple sample means,and Pearson and Spearman correlation analysis were used for correlation analysis.P<0.05 was considered statistically significant.Results 1.Compared with the control group,the proportion of CD4+CD25+CD127-Treg in peripheral blood and the levels of IL-10,TGF-β,IL-6,PGE-2 and HMGB1 in serum were significantly increased in esophageal squamous cell carcinoma patients(P<0.05),while the proportion of CD3+CD8+Teff in peripheral blood T lymphocytes and the level of IL-2 in serum were decreased(P<0.05)There were no significant differences in baseline Treg,Teff,serum IL-10,TGF-β,IL-6,PGE-2 and HMGB1 concentrations among patients with esophageal squamous cell carcinoma in different age,sex,lesion location and lesion length(P>0.05)but it is related to clinical stage(P<0.05).2.Compared with before and after radiotherapy,the proportion of Treg in peripheral blood,the levels of IL-10,TGF-β,IL-6,PGE-2 and HMGB1 in serum increased gradually(P<0.05),while the proportion of Teff and the level of IL-2 showed a decreasing trend(P<0.05).3.The number of Treg in peripheral blood of patients with esophageal squamous cell carcinoma was negatively correlated with the short-term efficacy(r=-0.478,P<0.05),the increased level of Treg cells after radiotherapy was negatively correlated with the short-term efficacy(r=-0.480,P<0.05),the number of Teff was positively correlated with the short-term efficacy(r=0.424,P<0.05),and the decreased level of Teff cells after radiotherapy was negatively correlated with the short-term efficacy(r=-0.491,P<0.05).4.There was a negative correlation between the number of Treg and the number of Teff,but there was no statistical significance(r=-0.201,P=0.288).The number of Treg was positively correlated with the contents of IL-10,IL-2,TGF-β and HMGBI in serum,without statistical significance(r=0.048,0.023,0.057,0.174,P=0.802,0.902,0.767,0.359,respectively).The number of Treg cells was positively correlated with the levels of PGE-2 and IL-6 in peripheral blood,and the difference was statistically significant(r was 0.497,0.410,P was 0.005,0.024).There was a negative correlation between the number of Teff and the number of Treg(r=-0.201,P=0.288),but there was no statistical significance.There was a negative correlation with IL-10,PGE-2,IL-2,and a positive correlation with IL-6,with no statistical significance(r=-0.053,-0.049,-0.031,0.015,P=0.780,0.796,0.871,0.939).And the levels of TGF-β and HMGB1 in peripheral blood were negatively correlated with statistical significance(r=-0.394,-0.449,P=0.031,0.013).Conclusion:1.The proportion of Treg cells to CD4+T cells in peripheral blood and the concentrations of IL-10,TGF-β,IL-6,PGE-2 and HMGB1 in serum of esophageal squamous cell carcinoma patients were higher than those of healthy subjects,while the proportion of Tefl cells to peripheral blood and the level of IL-2 in serum of esophageal squamous cell carcinoma patients were lower than those of healthy subjects.2.Routine clinical radiotherapy can increase the proportion of Treg cells in CD4+T cells and the concentrations of IL-10,TGF-β,IL-6,PGE-2 and HMGB1 in serum of patients with esophageal squamous cell carcinoma,reduce the proportion of Teff to peripheral blood T lymphocytes and the level of IL-2 in serum,thus further inhibiting the immune function.3.Treg cells in esophageal squamous carcinoma patients peripheral blood accounted for the proportion of CD4+T cells,T lymphocytes percentage Teff,IL-10 in serum,TGF-beta,IL-6,PGE-2,HMGB1,IL-2 concentration is associated with tumor stage.The higher levels of Treg cells accounted for the proportion of CD4+T cells,IL-10 in serum,TGF-beta,IL-6,PGE-2,HMGB1and the lower Teff lymphocyte percentage,the concentration of serum IL-2 is associated with the later stage,4.The proportion of Treg cells in peripheral blood of patients with esophageal squamous cell carcinoma was negatively correlated with the curative effect of radiotherapy.The higher the proportion of Treg cells in baseline,the worse the curative effect of radiotherapy.The Teff ratio is positively correlated with the short-term efficacy,and the higher the number,the better the efficacy.The decreasing range of Treg cells after radiotherapy is even negatively correlated with the efficacy.The significantly decreased range of Treg cells is the worse the efficacy,which can be used as a predictor of the efficacy of radiotherapy. |
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