本文選題：胃腫瘤　切入點：調(diào)節(jié)性T細胞　出處：《西南醫(yī)科大學》2017年碩士論文

【摘要】：目的:胃癌(Gastric cancer,GC)是世界上腫瘤相關性死亡的主要原因之一,而調(diào)節(jié)性T細胞(Treg cells)被認為是抑制抗腫瘤免疫促進腫瘤免疫耐受的主要因素之一。以往有關Treg細胞與胃癌的研究結果存在爭議,現(xiàn)已發(fā)現(xiàn)Treg細胞是一類具有異質(zhì)性的T細胞亞群,存在表型和功能的不同,這可能是導致之前結果不一致的原因。而目前鮮有關于這類Treg亞群細胞與胃癌的研究報道,因此本次研究通過檢測胃癌患者外周血以及腫瘤組織中Treg細胞及亞群的表達,探索Treg細胞及亞群與胃癌患者各臨床參數(shù)的相關性。方法:搜集胃癌患者和健康對照組外周血、同一患者的胃癌組織和癌旁正常組織,制備、分離、提純單個核細胞,標記表面抗體CD3、CD4、CD45RA、CD25和核內(nèi)抗體Foxp3。再根據(jù)CD45RA和Foxp3的表達將Treg細胞分為CD45RA+Foxp3lo(rTreg),CD45RA-Foxp3hi(aTreg)和CD45RA-Foxp3lo(non-Treg)三種亞群。流式細胞儀檢測分析各個組Treg及亞群細胞占CD4+細胞的比例。Graph Pad Prism統(tǒng)計軟件(5.0版本)統(tǒng)計分析數(shù)據(jù),Kruskal-wallis檢驗(多組)、Mann-Whitney U檢驗(兩組)比較各組間的差異是否具有統(tǒng)計學意義。結果:胃癌患者外周血(GC-PBMC)及腫瘤浸潤淋巴細胞(TIL)中總的CD4+CD25+Foxp3+Treg細胞占CD4+T細胞比例要比健康對照組(HD-PBMC)和癌旁正常組織浸潤淋巴細胞(NIL)的明顯增加(P0.0001,P0.0001),其差異有統(tǒng)計學意義。在外周血中胃癌患者的aTreg和non-Treg細胞比例較健康對照組明顯增加(P0.0001,P0.0001),差異有統(tǒng)計學意義。在組織浸潤淋巴細胞中,TIL-aTreg和TIL-non-Treg細胞比例較NIL明顯增加(P0.0001,P0.001)。分析Treg細胞及其亞群與各臨床病理參數(shù)關系發(fā)現(xiàn),僅發(fā)現(xiàn)腫瘤轉移(淋巴轉移和遠處轉移)與Treg及亞群細胞相關,而與腫瘤位置、大小、分化程度、分期以及血清CEA濃度無明顯相關性。出現(xiàn)轉移者無論在PBMC還是TIL中總的CD4+CD25+Foxp3+Treg細胞所占比例均較無轉移者明顯增加(P0.05,P0.01),亞群細胞中只有轉移患者TIL中的aTreg細胞所占比例較無轉移者明顯增加(P0.01)。結論:我們的研究證實了在胃癌患者外周血和TIL中Treg細胞所占比例明顯增加,以aTreg細胞增加最為明顯。而且只有TIL中增加的aTreg細胞與腫瘤轉移有關,提示在腫瘤微環(huán)境中起主要抑制抗腫瘤免疫的是TIL中的aTreg細胞,促使疾病的進展。所以在今后的免疫靶向治療中,局部地選擇性消除這類亞群細胞而非消除其他亞群細胞和(或)全身性的消除整個Treg細胞將有望成為一項有效治療胃癌的手段。同時我們還發(fā)現(xiàn)胃癌患者外周血及TIL中的non-Treg細胞也是增加的,但與疾病無明顯關聯(lián),所以在以后的研究中,我們需要鑒別不同的Treg亞型細胞而非整個Treg細胞與腫瘤的關系,這樣才有可能得出更為準確的結果。
[Abstract]:Objective: Gastric cancer is one of the major causes of tumor-related death in the world, and regulatory T cell line Treg cells is considered to be one of the main factors to inhibit anti-tumor immunity and promote tumor immune tolerance.Previous studies on Treg cells and gastric cancer have been controversial. It has been found that Treg cells are a class of heterogeneous T cell subsets with different phenotypes and functions, which may be the cause of previous inconsistent results.However, there are few reports on the expression of Treg cells and subsets in peripheral blood and tumor tissues of patients with gastric cancer.To explore the correlation between Treg cells and subsets and clinical parameters in patients with gastric cancer.Methods: the peripheral blood samples of patients with gastric cancer and healthy controls were collected, and the gastric cancer tissues and adjacent normal tissues of the same patients were collected. Mononuclear cells were prepared, isolated, purified, and labeled with surface antibody CD3p4, CD45RACD25 and intranuclear antibody Foxp3.According to the expression of CD45RA and Foxp3, Treg cells were divided into three subgroups: CD45RA Foxp3losrTrega (CD45RA-Foxp3hia Trega) and CD45RA-Foxp3losl (non-Tregt).Flow cytometry was used to analyze the ratio of Treg and subgroup cells to CD4 cells. The statistical analysis data were Kruskal-wallis test (Mann-Whitney U test).Results: the percentage of total CD4 CD25 Foxp3 Treg cells in peripheral blood of gastric cancer patients was significantly higher than that in healthy controls (HD-PBMC) and adjacent normal tissues (P 0.0001 / P 0.0001).The percentage of aTreg and non-Treg cells in peripheral blood of gastric cancer patients was significantly higher than that of healthy controls.The proportion of TIL-a Treg and TIL-non-Treg cells in infiltrating lymphocytes was significantly higher than that in NIL.By analyzing the relationship between Treg cells and their subsets and clinicopathological parameters, it was found that only tumor metastasis (lymphatic metastasis and distant metastasis) was associated with Treg and subgroup cells, but with tumor location, size and differentiation degree.There was no significant correlation between stages and serum CEA levels.The percentage of total CD4 CD25 Foxp3 Treg cells in both PBMC and TIL was significantly higher than that in patients without metastasis, and the percentage of aTreg cells in TIL of patients with metastasis was significantly higher than that of patients without metastasis.Conclusion: our study confirmed that the proportion of Treg cells in peripheral blood and TIL was significantly increased in gastric cancer patients, especially in aTreg cells.Moreover, only the increase of aTreg cells in TIL is related to tumor metastasis, suggesting that aTreg cells in TIL mainly inhibit anti-tumor immunity in tumor microenvironment, and promote the progression of the disease.Therefore, in the future, the local selective elimination of these subsets rather than the elimination of other subsets and / or the elimination of the whole Treg cell may be an effective method for the treatment of gastric cancer.We also found that the number of non-Treg cells in peripheral blood and TIL was increased in patients with gastric cancer, but there was no significant association with the disease, so in future studies, we need to distinguish the relationship between different Treg subtypes rather than the whole Treg cell.This makes it possible to arrive at more accurate results.
【學位授予單位】：西南醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.2

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相關期刊論文 前1條

1 Antoni M Szczepanik;Maciej Siedlar;Marek Sierzega;Dominika Goroszeniuk;Karolina Bukowska-Strakova;Antoni Czupryna;Jan Kulig;;T-regulatory lymphocytes in peripheral blood of gastric and colorectal cancer patients[J];World Journal of Gastroenterology;2011年03期

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本文編號：1723809