| Objective: This study aims to evaluate the expression of HMGB1,HMGN1,PD-1,PD-L1 and P16 in cervical tumor cells,and to discuss their relationships with the tumor-infiltrating lymphocytes in cervical carcinoma.To testify the expression of cytokines associated with Th1/Th2/Th9/Th17Th22,and compare the expression of immune checkpionts with immue response in peripheral blood.Methods:Paraffin embedded tumor tissue samples of 100 patients with cervical carcinoma were involved in this study.The immunohistochemical staining was applied to detect the expression of HMGB1,HMGN1,CD3,CD4,CD8 and FOXP3 in cervical tumors.HMGB1 and HMGN1 were divided into high expression and low expression on the basis of intensity,and into positive and negative group according to expression.PD-1,PD-L1 and P16 were devided into positive and negtive group on the basis of the percentage of positive cells.For the section of CD3,CD4,CD8 and FOXP3,the mean positive cells of five high magnification vision were acquire.50 cases of the peripheral blood from cancer patients were collected and separated,and we used ELISA to testify the serum expression of HMGB1,and multiplex cytokine detection systems to detect the expression levels of cytokines associated with Th1/Th2/Th9/Th17/Th22.Results: 1.The expression of HMGB1 and HMGN1 in tumor tissue of cervical cancer patients and its correlation with TILs:(1)No significant differences were observed between the expression of HMGB1 or HMGN1 and the patient age,FIGO stage as well as differentiation(P>0.05);(2)Cytoplasmic expression of HMGB1 and HMGN1 in tumor cells showed a significant positive correlation(P<0.05);(3)High levels of CD3+ and CD8+ expression emerged in tumors with cytoplasmic expression of HMGB1 or HMGN1 rather than in that without cytoplasmic expression of two proteins(P<0.05);(4)there was no significant relevance between cytoplasmic expression of HMGB1 and the number of CD4+ and Foxp3+ cells;(5)the number of CD4+ cells of HMGN1 positive group and negative group respectively were(249.71±110.01)and(205.24±126.76),with significant differences(P<0.05),but no significant differences were observed in Foxp3+ beteeen the two group(P>0.05).2.The expression of PD-1/PD-L1 in cervical cancer tissue:(1)the positive rate of PD-1 on lymphocytes and PD-L1 on tumor cells in tumor tissue respectively were 41% and 50%,meanwhile,there was no relevance between them and clinicopathologic variables;(2)no significant discrepancy in the number of CD3+,CD4+,CD8+ and Foxp3+ cells in tumor tissue bettween PD-1/PD-L1 positive and negative group were found(P>0.05);(3)nuclear and cytoplasmic HMGN1 expression in PD-1 positive cancer cells were lower than that in negative cancer cells(P<0.05).3.The expression of P16 in cervical tumor tissue:(1)the positive rate of P16 in tumor is 78 percent,the expression of P16 was not related to clinicopathologic variables,expression of HMGB1 and HMGN1;(2)the number of CD3+ and CD4+ T cells in P16 positive cells was more than that in negative cells(P<0.05),but there was no differences bettween the two group in the number of CD8+ and FOXP3+ T cells(P>0.05);(3)the expression of PD-L1 in P16 positive group was higher than latter,but the difference was no significant(P>0.05).4.The serum HMGB1 expression levels of cervical cancer patients:(1)the concentration of serum HMGB1 of II stage patients was(23.8048±5.1110)ng/ml,which was much higher than that of I stage patients(20.4743±6.3155)ng/ml(P<0.05),however,the level of HMGB1 was not related to age,differentiation,uniparous age and number of pregnancy and deliver(P>0.05).(2)the concentration of serum HMGB1 of patients with cytoplasmic HMGB1 expression was(23.7765±6.0113)ng/ml,which was much higher than that of patients without cytoplasmic HMGB1 expression(P<0.05),demonstrating that the serum HMGB1 levels of patients with cervical cancer correlated with cytoplasmic HMGB1 expression in tumor tissue.5.The levels of cytokines in peripheral blood:(1)the concentration of IL-6,IL-10 and IL-17 A of II stage patients was higher than that in I stage patients(P<0.05);(2)the Th1/Th2 cytokine ratios IFN-?/IL-5 was reversely correlated with the concentration of HMGB1 in peripheral blood of cervical cancer patients(P<0.05),TNF-?/IL-5 was positively correlated with PD-1 expressed on lymphocytes in tumor microenvironment(P<0.05),furthermore,TNF-?/IL-10 was negatively related to clinical stage(P<0.05).Conlusion: HMGB1 and HMGN1 secreted from cervical tumor cells participated in raising of TILs in tumor microenvironment.PD-1,PD-L1 and P16 did not take part in the progress of TILs movement in cervical cancer.HMGB1 secreted by cervical cancer cells maybe one crucial resource of serum HMGB1 of patients.Meanwhile,the serum HMGB1 levels of II stage patients was higher than that of I stage patient,illustrating that HMGB1 maybe involve in cervical cancer progression.The IL-6,IL10 and IL-17 A may involeved in the development of cervical cancer.HMGB1 in peripheral blood and PD-1 on lymphocytes in tumor microenvironment may take part in regulating the balance of Th1 or Th2 type of immune response. |
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