卵巢癌患者髓源性抑制细胞、调节性T细胞及其相关细胞因子的表达及临床意义

发布时间：2018-03-22 07:34

本文选题：卵巢癌　切入点：调节性T细胞　出处：《天津医科大学》2015年博士论文　论文类型：学位论文

【摘要】：目的作为常见妇科肿瘤之一,卵巢癌死亡率高,居妇科恶性肿瘤第一位。卵巢癌目前的治疗方案是以手术为主,术后辅助化疗、放疗、生物治疗等的综合治疗。由于70%左右的患者就诊时已于晚期,综合治疗后的患者仍有80%~85%左右会于短期内出现复发与转移,严重影响到卵巢癌患者治疗的疗效与预后,威胁着患者的生命。因此,寻找新的辅助治疗方法,提高或改善现有治疗手段的效果,以期实现遏制肿瘤转移和延缓肿瘤复发、提高生存率和改善生存质量的目的。多方研究报道表明,以往多种肿瘤免疫治疗方案效果不佳的原因可能与肿瘤免疫微环境相关,现在的研究主要集中在有关肿瘤免疫抑制微环境中免疫抑制细胞及其相关细胞因子在肿瘤免疫治疗中作用,这类研究希望能在不远的将来为肿瘤的治疗带来新的突破,也将为肿瘤免疫治疗效果不佳的现状提供新的解释。本研究通过检测卵巢癌外周血中CD33+HLA-DR-/low髓系来源抑制性细胞(myeloid-derived suppressor cells,MDSCs)、CD4+CD25+CD127-/low调节性T细胞(regulatory T cells,Tregs)的含量及卵巢癌患者血清中Th1、Th2类细胞因子的表达,分析这些肿瘤免疫微环境中的重要组成因素在卵巢癌患者中的表达特点,进一步分析它们与卵巢癌各项临床病理参数的关系,为深入揭示免疫抑制因素与卵巢癌进展及治疗的关系提供依据,为探索能够全面控制肿瘤免疫抑制微环境的有效措施提供实验基础。方法选择72例卵巢癌患者作为研究对象,30例健康人群作为对照组。采集清晨空腹外周血。应用流式细胞术检测外周血中CD33+HLA-DR-/low髓系来源抑制性细胞(myeloid-derived suppressor cells,MDSCs)和CD4+CD25+CD127-/low调节性T细胞(regulatory T cells,Tregs)的含量,分析二者在卵巢癌患者外周血中的表达特点,探讨二者与卵巢癌患者年龄、肿瘤大小、病理类型、病理分级、临床分期等各项临床病理参数的关系,探讨二者表达的相关性。应用BD CBA流式液相多重蛋白定量技术检测卵巢癌患者血清中Th1、Th2类细胞因子的表达及其与临床病理参数的关系。结果1.卵巢癌患者外周血中MDSCs为一群CD33阳性而HLA-DR阴性表达或低表达的细胞,与健康对照组(0.762±0.214)%相比,卵巢癌患者外周血中CD33+HLA-DR-/lowMDSCs的表达比例明显升高(3.391±1.931)%,差异有统计学意义(P0.01);其表达水平与卵巢癌患者的年龄、肿瘤大小、病理类型无关(P0.05),而与卵巢癌患者病理分级、临床分期具有明显联系,差异具有统计学意义(P0.05);与术前比较,手术后外周血中CD33+HLA-DR-/low MDSCs的表达水平(2.019±1.034)%明显降低,差异具有统计学意义(P0.05);2.卵巢癌患者外周血中Treg细胞为一群CD4和CD25双阳性而CD127低表达的细胞,平均百分比(14.068±4.546)%,较健康对照组外周血中(5.051±1.170)%显著增高,差异具有统计学意义(P0.01);Treg细胞的表达与卵巢癌患者的年龄、肿瘤大小、病理类型及肿瘤分化程度均无明显的相关性(P0.05),而与卵巢癌患者的临床分期密切相关,具有显著的统计学差异(P0.05);手术后外周血中CD4+CD25+CD127-/lowTregs的平均百分比为(12.243±4.213)%,较术前有所降低,差异具有统计学意义(P0.05);3.卵巢癌患者外周血中CD33+HLA-DR-/lowMDSCs的表达与CD4+CD25+CD127-/low Tregs的表达具有明显的相关性(n=72,r=0.0563,P0.05);4.卵巢癌患者血清中的Thl类细胞因子IL-2,IFN-γ的水平明显低于正常人(P0.01),而Th2类细胞因子IL-6、IL-10的水平显著高于正常人(P0.01),IL-4,TNF的水平与健康人比较无显著差异。且早期患者血清中IL-2,IFN-γ的水平明显高于晚期患者,而IL-6、IL-10的表达水平则低于晚期患者。结论卵巢癌患者外周血中免疫抑制性细胞CD33+HLA-DR-/low MDSCs和CD4+CD25+CD127-/lowTregs含量较健康对照均明显增高,二者的表达表现出明显的相关性,且Thl类细胞因子的水平降低,Th2类细胞因子的水平增高,提示卵巢癌患者体内免疫抑制状态的形成,为寻找卵巢癌独特的免疫调节机制以逆转卵巢癌免疫缺陷、改善卵巢癌预后提供了有力的实验依据和方法基础。
[Abstract]:Objective as one of the most common gynecological tumors, ovarian cancer mortality is high, ranking first in gynecological malignant tumor. Treatment of ovarian cancer is surgery, adjuvant chemotherapy, postoperative radiotherapy, biological therapy and other comprehensive treatment. Since about 70% of the patients have been in the late stage, after the comprehensive treatment of patients is about 80%~85% there will be recurrence and metastasis in the short term, seriously affect the curative effect and prognosis of patients with ovarian cancer treatment, threatening the lives of patients. Therefore, looking for new adjuvant treatment, improve or improve the existing treatment effect, in order to achieve curb tumor metastasis and delay the tumor recurrence and improve survival rate and improve the quality of life. Objective. Many reports indicated that the possible reasons for treatment scheme for multiple tumor immune effect and tumor immune microenvironment, now the research mainly focuses on the related tumor immunity Inhibition in the microenvironment of immunosuppressive cells and related cytokines in tumor immunotherapy, this kind of hope in the near future for cancer therapy bring new breakthrough, will also provide a new explanation for the status of tumor immunotherapy effect. This study through the detection of ovarian cancer in peripheral blood CD33+HLA-DR-/low myeloid derived suppressor cells (myeloid-derived suppressor cells, MDSCs), CD4+CD25+CD127-/low regulatory T cells (regulatory T cells, Tregs) and the content of Th1 in serum of patients with ovarian cancer, the expression of Th2 cytokines and expression analysis of characteristics in patients with ovarian cancer in these important factors in the tumor microenvironment, further analysis their relationship with clinicopathological parameters in ovarian cancer, and provide the basis for further revealing the relationship between progress and treatment of immunosuppressive factors and ovarian cancer, to explore the can Comprehensive and effective measures to control the immunosuppressive microenvironment provide experimental basis. Methods 72 cases of ovarian cancer patients as the research object, as a control group of 30 healthy people. Collected fasting peripheral blood. Flow cytometry was used to detect peripheral blood CD33+HLA-DR-/low myeloid derived suppressor cells (myeloid-derived suppressor cells, MDSCs) and CD4+CD25+CD127-/low regulatory T cells (regulatory T cells, Tregs) content, expression of the characteristics of the two in patients with ovarian cancer in the peripheral blood of two patients, age, ovarian cancer and tumor size, pathological type, pathological grade, clinical stage and relationship between clinicopathological parameters, to investigate the correlation between the expression of two the application of BD CBA flow liquid multiple protein quantitative detection of serum Th1 in patients with ovarian cancer, expression of Th2 cytokines and its relationship with clinicopathological parameters. Results 1. Ovarian cancer patients peripheral blood MDSCs for a group of CD33 positive and HLA-DR negative expression or low expression of the cells, and the healthy control group (0.762 + 0.214)% compared to the ratio of CD33+HLA-DR-/lowMDSCs expression in peripheral blood of patients with ovarian cancer were significantly higher (3.391 + 1.931)%, the difference was statistically significant (P0.01); the the expression level of ovarian cancer and the patient's age, tumor size, pathological type, and independent (P0.05) classification and pathology of patients with ovarian cancer, the clinical stage is obviously, the difference was statistically significant (P0.05); compared with the preoperative, the expression level of CD33+HLA-DR-/low in peripheral blood of MDSCs after surgery (2.019 + 1.034)% was decreased, the difference was statistically significant (P0.05); 2. patients with ovarian cancer Treg cells in peripheral blood for a group of CD4 and CD25 double positive and low expression of CD127 cells, the average percentage (14.068 + 4.546)%, compared with the healthy control group in peripheral blood (5.051 + 1.170)% Significantly, the difference was statistically significant (P0.01); Treg cells expressed in patients with ovarian cancer and age, tumor size, were no significant correlation between pathological type and tumor differentiation (P0.05), and correlated to the clinical stage of ovarian cancer patients is closely related to a statistically significant difference (P0.05); the average percentage after surgery CD4+CD25+CD127-/lowTregs in peripheral blood was (12.243 + 4.213)%, compared with the preoperative decreased, the difference was statistically significant (P0.05); the expression of CD33+HLA-DR-/lowMDSCs in peripheral blood of CD4+ CD25+CD127-/low and Tregs 3. in patients with ovarian cancer have significant correlation (n=72, r=0.0563, P0.05); Thl cytokines IL-2 4. ovary cancer patients in the serum, IFN- gamma levels were significantly lower than the normal people (P0.01), and Th2 cytokines IL-6, IL-10 levels were significantly higher than those in normal people (P0.01), IL-4, TNF level and the health of people and there is no explicit The differences in the serum of patients with early IL-2. And, IFN- gamma levels were significantly higher than those in patients with advanced, and IL-6, the expression level of IL-10 was lower than that in patients with advanced ovarian cancer patients. Conclusion the peripheral blood cells of CD33+HLA-DR-/low MDSCs immune suppression and CD4+CD25+CD127-/lowTregs content were significantly increased compared with healthy controls, the expression of the two genes show significant correlation Thl, and cytokine levels decreased, Th2 increased cytokine levels, suggesting that the formation of immune suppression in patients with ovarian cancer, ovarian cancer and unique for immune regulation mechanism to human ovarian cancer immunodeficiency, provided the experimental basis and method to effectively improve prognosis of ovarian cancer.

【学位授予单位】：天津医科大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R737.31

【共引文献】