【作者】 王晶；

【导师】 钟鸣；

【作者基本信息】 中国医科大学 ， 口腔基础医学， 2010， 硕士

【摘要】 目的本研究探讨肿瘤相关巨噬细胞(TAMs)在人成釉细胞瘤(ABs)组织中的浸润及对ABs组织血管生成的影响,探讨前列腺素2 (PGE2)受体EP3蛋白在ABs组织中的表达与意义。材料与方法实验标本来自中国医科大学附属口腔医院和第一临床医院2001-2007年存档蜡块,其中ABs101例(其中原发66例、复发35例),正常口腔黏膜26例。应用HE常规染色法,在镜下观察人ABs中的中性粒细胞,巨噬细胞,淋巴细胞,浆细胞的浸润表达情况；应用免疫组化SP法及CD68, CD34鼠抗人单克隆抗体,EP3兔抗人多克隆抗体,对TAMs,微血管密度(MVD), PGE2受体EP3蛋白进行检测,通过结果判定,统计学处理进行分析。结果1、应用HE常规染色法观察ABs临床病理切片,发现人ABs中淋巴细胞,中性粒细胞,巨噬细胞浸润明显,有少量的浆细胞的浸润表达,这些炎性细胞有的位于靠近肿瘤上皮团的间质内,有的直接浸润于肿瘤上皮团内。ABs炎性细胞计数明显高于正常口腔粘膜(P值分别为0.000,P<0.05),具有统计学意义。2、ABs巨噬细胞计数与微血管记数明显高于正常口腔粘膜(P值分别为0.000,0.003,P<0.05)。巨噬细胞计数与MVD之间呈正相关(r=0.194,P值为0.026,P<0.05)。3、ABs中EP3表达主要在胞浆,以中度阳性强阳性为主,位于牙源性上皮的外周柱状或立方状细胞及中心星网状层细胞浆中；EP3表达具有异质性,分化较好的角化细胞中表达为强阳性,而在原始胚胎细胞中表达往往呈阴性或弱阳性；EP3蛋白部分视野下核阳性率较高,占总数的60.38%(64／106)。在炎细胞、血管内皮细胞、纤维细胞胞浆中EP3也为阳性表达。正常粘膜中EP3主要表达在表层和棘浅层。人类原发ABs、复发ABs中都有EP3的明显表达,阳性率依次为复发ABs 85.71%、原发ABs 80.30%；正常口腔粘膜也有表达其阳性率为42.31%。采用秩和检验,EP3蛋白在ABs与正常口腔粘膜有显著差异(P=0.000,P<0.05)；原发组,复发组与正常黏膜组间有显著差异,具有统计学意义(P值分别为0.000,0.029,P<0.05)。结论1、人ABs中巨噬细胞,淋巴细胞,中性粒细胞浸润明显,有少量浆细胞的浸润表达。炎性细胞构成了人ABs的肿瘤微环境。2、人ABs组织中有明显的TAMs的浸润且与ABs的血管生成有关。3、人ABs中炎性介质EP3在蛋白质水平上高表达且明显高于正常粘膜。更多还原

【Abstract】 ObjectiveThis study was to investigate the influence of Tumor-associated macrophages (TAMs) infiltration in the human ameloblastomas(ABs), the correlation between TAMs and angiogenesis in ABs, to investigate the expression and significance of prostaglandin E2(PGE2) receptor EP3 in the ABs.Materials and MethdodsAll of the samples were selected from department of oral pathology, school of stomotology, China Medical University in the period of 2001 to 2007. Include: ameloblastomas (n=101, including 66 primary ABs,35 recurring ABs), normal oral mucosa(n=26).Using the HE convention dyeing, Under microscope observes the neutral granular cell, the macrophage,the lymphocyte, acidophilia granular cell, cytoplasmic cell in human ABs. TAMs, MCD and EP3 were detected by streptavidin peroxidase method and avidin-biotin peroxidose complex technique of immunohistochemistry using monoclonal mouse anti-human CD68 and CD34 antibody, polyclonal rabbit anti-human EP3 antibody.Results1. Observes the ABs clinical pathological section using the HE convention dyeing, discovers in person ABs the macrophage, the lymphocyte, the neutral granular cell obvious infiltration, has the few cytoplasmic cells’s infiltration expression, these inflammatory cell some infiltrations in near tumor mesenchymal, some direct infiltrations in tumor epidermis group. Compare the infiltration with the normal mucous membrane to be remarkable, the P value respectively is 0.000, has statistics significance.2. The mean macrophage counts in ABs was significantly higher than the counts in normal mucosa (P<0.05). The mean macrophage counts in primary ABs, recurring ABs were different. The mean microvessel counts in ABs were higher than that in normal mucosa (P<0.05). The mean microvessel were strongly correlated with TAMs counts in ABs (P<0.01).3. EP3 was expressed in the plasma of peripheral columnar or cuboidal cells and central polyhedral cells of odontogenic epithelium, most of which were moderately positive and strongly positive. The expressions of EP3 had heterogenicity.It was strong positive in well differentiated keratinocytes, while in primary embryonic cells, It was negative or weakly positive..EP3 protein expressed strongly in the nuclus of ABs, and its nuclear positive rate was 60.38%(64/106).The expressions of EP3 in the plasm of phlogotic cells, vascular endothelial cells and fibrocytes were also positive. In normal mucosas, EP3 was expressed in surface layer and the superficial layer of stratum spinosum. The immunohistochemical staining of EP3 was obviously expressed in human primary ABs, recurrent ABs. the positive rate was recurrent ABs (85.71%), primary Abs (80.30%).And EP3 also expressed in NOMs, the positive rate of which was 42.31%. Using Rank sum test, we found that there were significant differences between NOMs and Abs (P=0.000).Conclusions1. In human ABs the macrophage, the lymphocyte, the neutral granular cell obvious infiltration, has the few cytoplasmic cell’s infiltration expression. The inflammatory cell constituted the ABs tumor microenvironment.2. The expression of TAMs in ABs was stronger than in normal mucosas, and it may play a major role in the regulation of angiogenesis in ABs.3. The protein expressions of EP3 in ABs was higner than that of normal mucosas.更多还原