The Detection And Significance Of Transcription Factors And Cytokines Of Th17/Treg Cells In The Gastric Patients

Objective:To analyze the expresstion of Th17/ Treg in the patients with gastric cancers and observe the relationship between Th17/Treg and the clinicopathologic type,We detected the expression levels of transcription factor and cytokines of Th17/Treg cells in gastric patients.Method:1) Peripheral blood mononuclear cells(PBMC) were acquired from healthy volunteers,and cultured with PMA+Ionomycin for 6 hours. The goal gene were amplificated by PCR,and plasmid standard sample using in QRT-PCR were build by T-A clone.2) All samples including 70 patients and 40 healthy volunteers were collected from Jiangsu University affiliated hospital -Zhenjiang First Hospital.All patients were registered gender,age,location of cancer, clinicopathologic types.The transcription factor of Th17/Treg cells RORγt and FoxP3 were determined by quantitative real-time PCR (QRT-PCR),and the cytokines including IL-17,IL-6,IL-1β,TGF-βmRNA were detected with the same method.3) The correlation of the mRNA levels between Foxp3 and RORγt, TGF-βwere assessed by using bivariate correlation analysis. 4) The concentration of IL-23 and IL-10 in plasma of gastric cancer patients were detected by enzyme linked immunosorbent assay (ELISA).Results:1.The results of transcription factors1) Compared with healthy volunteers,gastric patients have higher mRNA levels of RORγt and FoxP3 in the PBMCs(P＜0.05).2) The ratio between FoxP3 mRNA and RORγt mRNA was higher in gastric patients with metastasis than in non-metastasis(P＜0.05),while there was no obvious difference between non-metastasis and health volunteers(P＞0.05).3) The ratio of FoxP3 mRNA compared with RORγt mRNA showed the higher value in poorly differentiated clinicopathologic type group than in well differentiated group(P＜0.05)4) The bivariate correlation analysis indicated that there was no obvious association between FoxP3 and RORγt in gastric patients with metastasis(R~2=0.1520).2.The results of Th17/Treg associated factors1) Compared with healthy volunteers,gastric patients had obvious higher mRNA levels of Th17 cells associated cytokine IL-17,IL-6, IL-1βand Treg cells associated cytokine TGF-βin the PBMC (P＜0.05).In addition,the predominant high level of TGF-βmRNA was found in metastasis group,but IL-17,IL-6,IL-1βwere not found(P＞0.05).2) The result of assessing relationship between these cytokines and gastric clinicopathologic types suggested that there was higher level of TGF-βmRNA in poorly differentiated gastric patients group (P＜0.05);but no obvious difference about IL-17,IL-6,IL-1βmRNA among all groups(P＜0.05).3) The cytokines levels in plasma detected by ELISA indicated both IL-23 and IL-10 were higher in gastric patients than in healthy volunteers(P＜0.05);In addition,IL-10 was significant higher in metastasis group than non-metastasis group(P＜0.05);but IL-23 was not found(P＞0.05).Conclusions:1) There was higher expression of Th17/Treg cells in PBMC of gastric cancer patients.2) The predominant expresstion of Treg was found in advanced gastric patients,and in poorly differentiated group,while Th17 cells only had higher level in gastric patients,but seems not significant change between non-metastasis and metastasis groups.3) The expresstion character of Th17/Treg may related to their cytokines regulation in immue internal environment.