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

【摘要】：目的:胃癌(Gastric cancer,GC)是世界上腫瘤相關(guān)性死亡的主要原因之一,而調(diào)節(jié)性T細(xì)胞(Treg cells)被認(rèn)為是抑制抗腫瘤免疫促進(jìn)腫瘤免疫耐受的主要因素之一。以往有關(guān)Treg細(xì)胞與胃癌的研究結(jié)果存在爭(zhēng)議,現(xiàn)已發(fā)現(xiàn)Treg細(xì)胞是一類具有異質(zhì)性的T細(xì)胞亞群,存在表型和功能的不同,這可能是導(dǎo)致之前結(jié)果不一致的原因。而目前鮮有關(guān)于這類Treg亞群細(xì)胞與胃癌的研究報(bào)道,因此本次研究通過檢測(cè)胃癌患者外周血以及腫瘤組織中Treg細(xì)胞及亞群的表達(dá),探索Treg細(xì)胞及亞群與胃癌患者各臨床參數(shù)的相關(guān)性。方法:搜集胃癌患者和健康對(duì)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(gè)核細(xì)胞,標(biāo)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(dá)將Treg細(xì)胞分為CD45RA+Foxp3lo(rTreg),CD45RA-Foxp3hi(aTreg)和CD45RA-Foxp3lo(non-Treg)三種亞群。流式細(xì)胞儀檢測(cè)分析各個(gè)組Treg及亞群細(xì)胞占CD4+細(xì)胞的比例。Graph Pad Prism統(tǒng)計(jì)軟件(5.0版本)統(tǒng)計(jì)分析數(shù)據(jù),Kruskal-wallis檢驗(yàn)(多組)、Mann-Whitney U檢驗(yàn)(兩組)比較各組間的差異是否具有統(tǒng)計(jì)學(xué)意義。結(jié)果:胃癌患者外周血(GC-PBMC)及腫瘤浸潤(rùn)淋巴細(xì)胞(TIL)中總的CD4+CD25+Foxp3+Treg細(xì)胞占CD4+T細(xì)胞比例要比健康對(duì)照組(HD-PBMC)和癌旁正常組織浸潤(rùn)淋巴細(xì)胞(NIL)的明顯增加(P0.0001,P0.0001),其差異有統(tǒng)計(jì)學(xué)意義。在外周血中胃癌患者的aTreg和non-Treg細(xì)胞比例較健康對(duì)照組明顯增加(P0.0001,P0.0001),差異有統(tǒng)計(jì)學(xué)意義。在組織浸潤(rùn)淋巴細(xì)胞中,TIL-aTreg和TIL-non-Treg細(xì)胞比例較NIL明顯增加(P0.0001,P0.001)。分析Treg細(xì)胞及其亞群與各臨床病理參數(shù)關(guān)系發(fā)現(xiàn),僅發(fā)現(xiàn)腫瘤轉(zhuǎn)移(淋巴轉(zhuǎn)移和遠(yuǎn)處轉(zhuǎn)移)與Treg及亞群細(xì)胞相關(guān),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(guān)性。出現(xiàn)轉(zhuǎn)移者無論在PBMC還是TIL中總的CD4+CD25+Foxp3+Treg細(xì)胞所占比例均較無轉(zhuǎn)移者明顯增加(P0.05,P0.01),亞群細(xì)胞中只有轉(zhuǎn)移患者TIL中的aTreg細(xì)胞所占比例較無轉(zhuǎn)移者明顯增加(P0.01)。結(jié)論:我們的研究證實(shí)了在胃癌患者外周血和TIL中Treg細(xì)胞所占比例明顯增加,以aTreg細(xì)胞增加最為明顯。而且只有TIL中增加的aTreg細(xì)胞與腫瘤轉(zhuǎn)移有關(guān),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(xì)胞,促使疾病的進(jìn)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(xì)胞而非消除其他亞群細(xì)胞和(或)全身性的消除整個(gè)Treg細(xì)胞將有望成為一項(xiàng)有效治療胃癌的手段。同時(shí)我們還發(fā)現(xiàn)胃癌患者外周血及TIL中的non-Treg細(xì)胞也是增加的,但與疾病無明顯關(guān)聯(lián),所以在以后的研究中,我們需要鑒別不同的Treg亞型細(xì)胞而非整個(gè)Treg細(xì)胞與腫瘤的關(guān)系,這樣才有可能得出更為準(zhǔn)確的結(jié)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(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.2

【參考文獻(xiàn)】

相關(guān)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(hào)：1723809