| Objective：to detect the expression levels of Th17cells and Treg cells in colon cancertumor tissues of patients and the expression levels of related cytokines in the peripheralblood, analysis of Th17/Treg in patients with colon cancer changes, observe therelationship with its clinical and pathological tissue typing, and discussed the relationshipwith the tumor occurrence and development and its possible mechanism.Methods：Patients with colon cancer (n=28) were divided into two groups: those withearly disease (n=8) corresponding to Dukes A according to the Dukes classification forcolon cancer and those with advanced disease corresponding to Dukes B, C and D (n=20).None of the patients received radiotherapy, chemotherapy，or other medical interventionsduring this study. Peripheral blood mononuclear cells (PMBC)were obtained from thepatients (n=20) and healthy volunteers (n=20). Tumor-infiltrating lymphocytes (TILs) andthe normal colonic mucosa as controls were isolated from resected tumor specimens frompatients with advanced colonic cancer (n=28). The frequencies of Thl7cells and Treg cellsas a percentage of total CD4~+T cells in TILs and normal colonic mucosa were evaluated byfluorescence-activated cell sorting (FACS) analysis.The expression level of Thl7celland Treg cell in TILs and normal colonic mucosa were investigated byimmunohistochemical technique (Immunohistochemistry,IHC). The transcription factorexpression level of Th17/Treg cells-related cytokines (IL-23, IL-10) in the peripheral bloodof colorectal cancer patients were detected using reverse transcription polymerase chainreaction (RT-PCR).Results：1. In TILs from patients with early disease of colon cancer patients, the frequencyof Th17cells and Treg cells was significantly higher than that in intra-epithelial lymphocytes (IELs) of normal colonic mucosa in the same cohorts (8.38±2.86%VS3.56±1.26%and7.13±1.65%VS2.74±1.09%,P<0.05); In TILs from patients withadvanced disease, the frequency of Th17cells and Treg cells was also significantly higherthan that in IELs of normal colonic mucosa in the same cohorts (9.23±1.37%VS4.82±1.56%and16.57±3.78%VS3.45±1.67%,P<0.05); In TILs from patients withadvanced disease the frequency of Treg cells was significantly higher than earlydisease(16.57±3.78%VS7.13±1.65%,P<0.05); but in advanced colorectal carcinoma cellTh17cells was not obvious differences than early disease (9.23±1.37%VS8.38±2.86%,P>0.05),while the ratio of Th17/Treg cells In TILs from patients with advanced diseasewas significantly lower than early disease.2. Immunohistochemical detection of colorectaltumors and normal tissues of Thl7cell and Treg cell expression levels, the results showthat: the early colon cancer tumor tissue within the number of Th17positive cells and thenumber of Treg positive cells was significantly higher than that of early normal tissuesadjacent to carcinoma(17.2±1.3VS2.6±3.6and16.5±2.4VS3.8±2.5,P<0.01); advancedcolorectal tumor tissue within the number of Th17positive cells and Treg positive cellnumber was significantly higher than that in advanced normal tissues adjacent tocarcinoma(19.3±2.8VS2.8±2.1and30.3±3.2VS4.1±1.2,P<0.01); early colorectal tumortissue in Th17positive cell number was not obvious differeces than advanceddisease(17.2±1.3VS19.3±2.8,P>0.05); and early colon cancer tumors in number of Tregpositive cells was significantly lower than advanced disease(16.5±2.4VS30.3±3.2,P<0.05).3. The results of PCR showed: IL-23, IL-10was highly expressed inperipheral blood of colon cancer patients with healthy control group (P<0.05), andIL-10mRNA expression levels was significant difference between advanced disease andearly disease (0.98±0.07%VS0.62±0.05%,P<0.05), while IL-23between the two groupshad no significant difference (0.72±0.02%VS0.68±0.03%, P>0.05).Conclusion： The results indicate that with disease progression, Th17cells and Treg cellsin TILs is gradually increased, and Treg cells than Th17cells increased more obvious.Cytokine plays an important role in Th17cells and Treg cell differentiation and development, and the trend of changes of related cytokines IL-23and IL-10in theperipheral blood of colon cancer was the agreement with that of Th17cells and Treg cellsin tumor tissue. The Th17cells and Treg cells in colon cancer may be associated withtumor immunity in environment related cytokine regulation. Further revealed that the Thl7/Treg balance adjustment mechanism may be provided a theoretical and experiment basisfor colon cancer immune therapy. |
PDF Full Text Request