| Objective: To detect the expression of carbonic anhydrase-IX(CA-IX)and epithelial cadherin(E-cad)in tissues of elderly patients with locally advanced non-small cell lung cancer(NSCLC)and normal lung,and the percentage of regulatory T cells(Treg cells)in peripheral blood of NSCLC patients and healthy people.The relationships between the three and clinical-pathological characteristics of NSCLC and correlation between each other were analyzed,and their impact on the radiotherapy efficacy and prognosis were explored further.To provide more theoretical basis to find effective markers of predicting the curative effect and estimating prognosis for elderly patients with locally advanced NSCLC with the individual treatment.Methods:1 A retrospective study was applied to collect paraffin blocks of cancer tissues from 53 cases of patients with locally advanced NSCLC and 16 cases of normal lung tissues from Department of Pathology,Hebei General Hospital from June 2011 to June 2014.53 cases of patients with NSCLC were not operated,diagnosed by biopsying with fiberbronchoscopy or percutaneous lung biopsy,KPS?70,age?60 and treated with no chemotherapy but only radiotherapy.Other 12 cases of peripheral blood samples from healthy people were collected.2 Expression of CA-IX and E-cad in 53 cases of locally advanced NSCLC tissues and 16 cases of normal lung tissues were detected by immunohistochemistry(SP method)and flow cytometry was used to detect the percentage of Treg cells in peripheral blood of 53 cases of patients with NSCLC and 12 cases of healthy people.3 All analyses were performed using SPSS version 13.0.The enumeration data were analyzed using the Chi-square test.Using mean±standard deviation to represent the measurement data,the comparison of the two samples were compared with t or t' test;Correlation analysis was performed using Spearman or linear correlation analysis.The Kaplan-Meier survival curves were used to analyse patient's survival time,and statistical significance was analyzed using the log-rank test.Independent factors of prognosis were studied by Cox regression model.All statistical tests were two-sided,and statistical significance was accepted at the P<0.05 level.Results:1 Expression of CA-IX protein in elderly patients with locally advanced NSCLC and the relationships with clinical pathological characteristics,radiotherapy response and prognosisExpression of CA-IX protein was mainly observed in the cancer cells with cell membrane or cytoplasm,and took a brownish yellow granular.The positive rates of CA-IX expression in NSCLC tissues and normal lung tissues were 64.2%(34/53)and 0%(0/16),there was significant difference between two groups(P<0.05).The results showed that there was no significance difference between the expression of CA-IX protein with gender,age and pathology type(P>0.05),while significantly correlated with TNM stage and differentiation degree(P<0.05).The positive expression of CA-IX protein in patients with IIIA stage and IIIB stage were 44.4%(8/18)and 74.3%(26/35),respectively.The positive expression of CA-IX protein in patients with high and middle differentiation,bad differentiation was 48.4%(15/31)and 86.4%(19/22),respectively.No cases CR and 17 cases PR in the positive expression of CA-IX of 34 cases,total effective rate was 50.0%(17/34);1 cases CR and 14 cases PR in the negative expression of CA-IX protein of 19 cases,total effective rate was 78.9%(15/19),and a significant difference was observed between the two groups(P<0.05).The analysis of statistical data showed there was probably a negative correlation between expression of CA-IX protein and the radiotherapy effect.With Kaplan-Meier survival curve analysis,the 1-,2-and 3-year survival rate of the patients with positive and negative expression of CA-IX were 70.4%,28.8%,6.4% and 73.7%,39.7%,17.0%,respectively.Difference was statistically significant by log-rank test(P<0.05),data showed that the expression of CA-IX protein was negatively correlated with the survival time of patients with locally advanced NSCLC.2 Expression of E-cad protein in elderly patients with locally advanced NSCLC and the relationships with clinical pathological characteristics,radiotherapy response and prognosisExpression of E-cad protein was mainly observed in the cancer cells with cell membrane,and took a brownish yellow granular.The positive rates of E-cad expression in NSCLC tissues and normal lung tissues were 43.4%(23/53)and 93.8%(15/16),there was significant difference between two groups(P<0.05).The results showed that there was no relationships between the E-cad expression and gender,age and pathology type(P>0.05),but significantly correlated with TNM stage and differentiation degree(P<0.05).The positive expression of E-cad protein in patients with IIIA stageand IIIB stage were 66.7%(12/18)and 31.4%(11/35),respectively.The positive expression of E-cad protein in patients with high and middle differentiation,bad differentiation were 58.1%(18/31)and 22.7%(5/22),respectively.1 cases CR and 17 cases PR in the positive expression of E-cad of 23 cases,total effective rate was 78.3%(18/23);No cases CR and 14 cases PR in the negative expression of E-cad of 30 cases,total effective rate was 46.7%(14/30);and the difference between two groups was statistical significance(P<0.05).The analysis of statistical data showed there was probably a positive correlation between expression of E-cad protein and the radiotherapy effect.With Kaplan-Meier survival curve analysis,the 1-,2-and 3-year survival rate of the patients with positive and negative expression of E-cad were 77.6%,43.8%,19.5% and 66.7%,24.6%,3.5%,respectively.Difference was statistically significant by log-rank test(P<0.05),data showed that the expression of E-cad protein was positively correlated with the survival time of patients with locally advanced NSCLC.3 The level of Treg cell in peripheral blood of elderly patients with locally advanced NSCLC and the relationships with clinical pathologic characteristics,radiotherapy outcome and prognosisThe ratio of Treg cells cells in the peripheral blood of NSCLC was(8.14±1.24)%,which was significantly higher than that in healthy control group(2.38±0.38)%,and the difference was statistically significant(P<0.05).The level of Treg cells in the peripheral blood of NSCLC patients was no significant difference in different gender,age and pathological types(P>0.05).Treg cells ratio were respectively(7.63±1.07)% and(8.85±1.11)% in high and middle differentiation,bad differentiation group;in the IIIA,IIIB group were respectively(7.46±1.30)%,and(8.49±1.06)%,and the difference was statistically significant(P<0.05).By Spearman rank test and linear correlation analysis,there was significantly negative correlation between the levels of Treg cells in peripheral blood of patients with NSCLC and radiotherapy efficacy and prognosis(r=-0.594,P=0.000;r=-0.989,P=0.000),namely the level of Treg cells was higher,the radiation effect was worse,survival time was shorter.4 The relationships between the expression of CA-IX and E-cad protein and the level of Treg cells in peripheral blood in elderly patients with locally advanced NSCLCWith Spearman rank test,there was a negative correlation between the expression of CA-IX and E-cad protein in the 53 cases of NSCLC tissues(r=-0.536,P=0.000).The levels of Treg cells in groups of positive and negative expression of CA-IX protein were respectively(8.38±1.16)% and(7.71±1.28)%,the difference was statistically significant(P<0.05),and data showed there was a positive correlation between the level of Treg cells and the expression of CA-IX protein.The levels of Treg cells in groups of positive and negative expression of E-cad protein were respectively(7.77±1.36)% and(8.42±1.07)%,the difference was no significant(P>0.05),suggesting there was no correlation between the levels of Treg cells and E-cad expression.5 The analysis of prognosis factors for elderly patients with locally advanced NSCLCAnalysis with one-way: Gender,age and pathology type were not correlated with the prognosis of NSCLC(P>0.05).But TNM stage,differentiation degree,CA-IX and E-cad,the level of Treg cells in peripheral blood were correlated with the prognosis of NSCLC(P<0.05).TNM stage,differentiation degree and CA-IX expression were independent prognostic factors of elderly patients with locally advanced NSCLC by Cox regression analysis.Conclusions:1 The expression of CA-IX protein in carcinoma tissues of elderly patients with locally advanced NSCLC is significantly higher than normal lung tissues.The expression of E-cad protein in carcinoma tissues is lower than normal lung tissues.The level of Treg cells in peripheral blood of elderly patients with locally advanced NSCLC is significantly higher than that of healthy people.And their expressions are related to TNM stage and differentiation degree.There is a negative correlation between the expression of CA-IX and E-cad protein,and the expression of CA-IX protein is probablypositively correlated with the level of Treg cells.2 There are significantly negative correlations between CA-IX expression and levels of Treg cells and radiotherapy effect and prognosis in elderly patients with locally advanced NSCLC,indicating that the prognosis is probably affected by tumor acidic microenvironment and immune status,and it is expected to become the index to judge the effect of radiotherapy and prognosis that CA-IX and Treg are detected together. |
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