| Part I The expression of PD-1 and analysis of lymphocyte subgroup in peripheral blood of NSCLCObjective:Explore the expression characteristics of lymphocyte subgroup and programmed death-1(PD-1) in the peripheral blood between the NSCLC patients including different stages,pathological types and the healthy controls,analyzing the correlation of the proportion of lymphocyte subgroup and PD-1 with the clinical stage,discussing its clinical significance.Methods:Thirty-eight cases of adenocarcinoma,thirty-five cases of squamous cell carcinoma of NSCLC patients in various cancer types and different clinical stages and thirty healthy controls was selected.The CD4+,CD8+,CD19+,NK cells in the peripheral blood can be detected by the FCM.The concentration of PD-1 in the peripheral blood was measured by ELISA.Results:(1) The expression level of PD-1 of Group A,B,C is(104.98±20.76),(109.13±24.65),(50.80±14.38)pg/m L,and the result of experimental group was higher than that of the healthy controls(F=78.581,P<0.001),having no statistical significance between group A and Group B;(2)Contrasting with the healthy controls,The positive expression ratio of CD4+,NK cells was lower and CD8+ cells was higher in the peripheral blood of the experimental group,which both had statistical significance(P<0.001),but the difference of CD19+ between them was little(P>0.05),both having no statistical significance between Group A and Group B(P>0.05);(3)Based on the staging criteria of TNM and contrasting the Stage I-II with Stage III-IV,the CD4+ cell-ratio was significantly lower(tA=7.329, tB=5.376),the expression level of PD-1 was significantly higher(tA=-6.151, tB=-6.601),the NK cell-ratio was also significantly lower(tA=3.772,tB=5.169), the CD8+ cell-ratio was significantly higher(tA=-0.592,tB=-6.416),which all had statistical significance(P<0.001),but the CD19+cell-ratio had no statistical significance(tA=-0.981,tB=-0.224,P>0.05);(4)The expression level of PD-1 had negative correlation with the ratio of NK cells(rA=-0.568,rB=-0.631),had positive correlation with the ratio of CD8+cells(rA=0.643,rB=0.648),which both had statistical significance(P<0.001).But the expression level of PD-1 had no correlation with the ratio of CD4+CD19+cells,having no statistical significance(P>0.05).Conclusion:Compared with the healthy controls,The immune function of experi-mental group was significantly lower,the positive expression ratio of PD-1 in the peripheral blood was higher,having certain corrections between them,thus both the detection of lymphocyte subgroup and PD-1 expression level playing a crucial role in the development of NSCLC and maybe becoming a method of screening immunotherapy of NSCLC. Part Ⅱ Relationship between the expression of PD-1 and the clinical pathological parameters in non-small cell lung cancerObjective:To research the expression characteristics of programmed death-1(PD-1) on pathological tissue and PD-1+CD8+T in the peripheral blood,thus analyzing the correlation and its relationship with clinicopathological parameters(age,gender,pathological type,TNM stages,the degree of differentiation, lymphatic metastasis and distant metastasis).Methods:Ninety cases of NSCLC pathological tissues and corresponding fresh blood was selected,with 45 cases of para-carcinoma tissue samples and 30 cases of peripheral blood of healthy controls for comparison.SP immune-histochemical method was used to research the expression of the PD-1 and FCM was used to study the expression of the PD-1+CD8+T.Results:(1) PD-1 is mainly expressed in tumor interstitium lymphocyte membrane,whose positive rate was 75.56%(68/90) and 6.67%(3/45), the difference was statistically significant(χ2=57.102, P<0.05);(2)The positive expression ratio of PD-1+CD8+T cell of the two groups was 16.73±3.93% and 7.92±2.00%,the difference was statistically significant(t=11.764,P<0.05);(3) The results were significantly correlated with each other(rs=0.780,P<0.001),which both related to the TNM stages,the degree of differentiation,lymphatic metastasis and distant metastasis(P<0.05),no obvious correlation with age, gender and pathological type(P>0.05).Conclusion:The expression of PD-1 was increased in NSCLC patients,and PD-1 signaling pathway may inhibit anti-tumor immunity.The expression of PD-1 is associated with the TNM stages,the degree of differentiation,lymphatic metastasis and distant metastasis of patients,which may be a proposing marker and indicators in NSCLC treatment.The test of PD-1+CD8+T in the peripheral blood may instead of the test of PD-1 in pathological tissue, which may be a indicator in screening patients who can targeted therapy of PD-1. |
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