HMGB1与肿瘤相关巨噬细胞在卵巢癌淋巴管生成方面的相关性研究

发布时间：2018-07-10 11:43

本文选题：HMGB1 + 肿瘤相关巨噬细胞　；参考：《天津医科大学》2014年硕士论文

【摘要】：目的：肿瘤微环境是一个由肿瘤细胞、基质细胞、免疫细胞以及细胞因子等共同构成的肿瘤局部复杂的炎性环境。大量研究已经证实肿瘤微环境中大量的细胞因子以及免疫炎性反应,对肿瘤的发生、进展等产生重要影响。HMGB1与肿瘤相关巨噬细胞作为肿瘤局部微环境的重要组成部分,对肿瘤的淋巴管生成以及后续的淋巴结转移有重要作用。同时,HMGB1作为外源性刺激因子,能作用于巨噬细胞进一步发挥其细胞学效应。该研究旨在探讨二者在促进肿瘤淋巴管生成方面有无相关性。方法：(1)免疫组化(S-P法)分别检测上皮性卵巢癌以及正常卵巢组织中HMGB1、肿瘤相关巨噬细胞(TAMs)及淋巴管密度(LVD)的表达。(2)用Spearman等级相关分析对上皮性卵巢癌患者HMGB1表达与LVD进行相关性分析,用Pearson相关分析对上皮性卵巢癌患者CD68阳性肿瘤相关巨噬细胞计数与LVD进行相关性分析。(3)MACS技术获取上皮性卵巢癌患者腹水中TAMs,设立对照组与实验组,对照组为未处理的LECs；实验组：分别用rHMGB1(2μg/ml)、CD14阳性肿瘤相关巨噬细胞培养上清、rHMGB1(2μg/ml)预处理的CD14阳性肿瘤相关巨噬细胞培养上清以及rVEGF-C(5ng/ml)处理的LECs作为实验组,通过CCK-8实验、细胞迁移实验、成管实验观察各组之间在淋巴管内皮细胞增殖、迁移、成管能力之间的差异。结果：(1)免疫组化结果显示：上皮性卵巢癌组织中HMGB1的表达和肿瘤相关巨噬细胞的浸润明显高于正常卵巢组织,两组之间的差异有显著统计学意义,且与淋巴结转移相关。(2)用Spearman等级相关分析对上皮性卵巢癌患者HMGB1表达与LVD进行相关性分析,可见上皮性卵巢癌患者中HMGB1表达与LVD成正相关(r=0.491,P0.001)。用Pearson目关分析对上皮性卵巢癌患者肿瘤相关巨噬细胞与LVD进行相关性分析,提示上皮性卵巢癌患者肿瘤相关巨噬细胞计数与LVD成明显正相关(R2=0.242,P0.001)。(3)体外实验结果提示：HMGB1与肿瘤相关巨噬细胞均可以促进淋巴管内皮细胞增殖、迁移、成管,然而,当二者相互作用后,这种促进淋巴管内皮细胞增殖、迁移、成管的能力大大增强。结论：上皮性卵巢癌中组织中HMGB1的表达和肿瘤相关巨噬细胞的浸润明显高于正常卵巢组织,且与肿瘤淋巴结转移密切相关。HMGB1与肿瘤相关巨噬细胞均可以促进淋巴管内皮细胞增殖、迁移、成管,然而,当二者相互作用后,这种促进淋巴管内皮细胞增殖、迁移、成管的能力大大增强,这将为抗卵巢癌淋巴管生成的靶向治疗提供理论依据。
[Abstract]:Objective: tumor microenvironment is a complex inflammatory environment composed of tumor cells, stromal cells, immune cells and cytokines. A large number of studies have confirmed that a large number of cytokines and immune inflammatory reactions in tumor microenvironment have important effects on tumor occurrence and progression. HMGB1 and tumor-associated macrophages are important components of tumor local microenvironment. It plays an important role in lymphangiogenesis and subsequent lymph node metastasis. At the same time, HMGB1, as an exogenous stimulant factor, can further exert its cytological effect on macrophages. The aim of this study was to investigate whether there is a correlation between the two in promoting lymphangiogenesis in tumors. Methods: (1) Immunohistochemistry (S-P method) was used to detect the expression of HMGB1, tumor associated macrophages (TAMs) and lymphatic vessel density (LVD) in epithelial ovarian carcinoma and normal ovarian tissues, respectively. (2) Spearman grade correlation analysis was used to detect HMGB1 expression in epithelial ovarian cancer patients. Da and LVD were analyzed. Pearson correlation analysis was used to analyze the correlation between CD68 positive tumor associated macrophage count and LVD in epithelial ovarian cancer patients. (3) Macs was used to obtain TAMsin ascites of patients with epithelial ovarian cancer. The control group and experimental group were set up and the control group was untreated LECs. Experimental group: LECs pretreated with rHMGB1 (2 渭 g/ml) tumor associated macrophage culture supernatant and rVEGF-C (5ng/ml) treated with rHMGB1 (2 渭 g/ml) were used as experimental group. The proliferation, migration and ability of lymphatic endothelial cells were observed. Results: (1) Immunohistochemical results showed that the expression of HMGB1 and the infiltration of tumor associated macrophages in epithelial ovarian carcinoma were significantly higher than those in normal ovarian tissues, and the difference between the two groups was statistically significant. (2) the expression of HMGB1 in epithelial ovarian cancer was correlated with LVD by Spearman rank correlation analysis. The positive correlation between HMGB1 expression and LVD was found in epithelial ovarian cancer patients (r 0.491p0.001). The correlation between tumor associated macrophages and LVD in patients with epithelial ovarian cancer was analyzed by Pearson muzzle analysis. These results suggest that the number of tumor-associated macrophages in epithelial ovarian cancer patients is positively correlated with LVD (R2O0.242P0.001). (3). The results of in vitro experiments indicate that both the tumor associated macrophages and the tumor associated macrophages can promote the proliferation, migration and angiogenesis of lymphatic endothelial cells. When the two interact, the ability to promote the proliferation, migration and angiogenesis of lymphatic endothelial cells is greatly enhanced. Conclusion: the expression of HMGB1 and the infiltration of tumor-associated macrophages in epithelial ovarian carcinoma are significantly higher than those in normal ovarian tissues. HMGB1 and tumor-associated macrophages can promote the proliferation, migration and angiogenesis of lymphatic endothelial cells. However, when the two interactions, this can promote the proliferation and migration of lymphatic endothelial cells. The ability of angiogenesis is greatly enhanced, which will provide a theoretical basis for targeted therapy against lymphangiogenesis of ovarian cancer.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.31

【参考文献】