【作者】 宋魏；

【导师】 仇晓菲；

【作者基本信息】 天津医科大学 ， 病理学与病理生理学， 2008， 硕士

【摘要】 背景韧带样型纤维瘤病（Desmoid-type fibromatosis，DTF），也称侵袭性纤维瘤病，是发生于深部软组织的克隆性纤维母细胞增生性肿瘤，具有浸润性生长、局部复发但不转移的特点。其治疗主要通过手术扩大切除及术后辅以适当的放疗和／或化疗，但术后局部复发率仍很高，需要多次手术。分子靶向治疗作为肿瘤治疗的新手段已显示其重要的临床价值。AnnexinⅡ是新近发现的纤溶酶原（plasminogen，PLG）和组织纤溶酶原激活物（tissue plasmin activator，tPA）的共同受体，涉及乳腺癌等实体肿瘤的转移。最近发现AnnexinⅡ可以和靶向药物血管抑素结合，可能是血管抑素抑制肿瘤进展的重要靶点。TGF-β₁是一种多功能细胞因子，在调节细胞增殖、分化和凋亡等方面发挥着重要作用。我们先前的研究表明TGF-β₁参与结节性筋膜炎的发病，发现其除涉及肌纤维母细胞的分化，可能还参与血管形成。检索Pubmed和万方数据库（1980-2008年）均未见DTF中AnnexinⅡ的研究；虽然已有个别文献报道TGF-β₁在DTF中的表达，但它在DTF发病中的作用尚不清楚。目的：1．了解AnnexinⅡ是否参与DTF的发病，进一步了解其是否与DTF的侵袭性相关，为今后临床开展血管抑素靶向治疗DTF提供实验依据。2．证实TGF-β₁在DTF中表达，进一步了解其是否与DTF中肿瘤细胞分化、间质构建及血管新生相关。方法：选取手术切除后石蜡包埋DTF 34例为研究对象，应用免疫组织化学染色方法，分别检测（1）AnnexinⅡ在DTF和周围正常组织中的表达及分布，进一步分析其在DTF浸润区和非浸润区的表达及分布；（2）TGF-β₁和TGF-βRI在DTF和周围正常组织中的表达及分布；（3）用α-SMA标记肌纤维母细胞，用CD31标记肿瘤血管，观察分析TGF-β₁／TGF-βRI与肌纤维母细胞分化、fibronectin表达和MVD的关系；并检测VEGF在DTF中的表达，分析它与MVD的关系。结果：1．AnnexinⅡ在DTF中的表达主要分布于肿瘤细胞和内皮细胞，阳性率为85．3％（29／34），阳性表达在浸润区尤为突出。AnnexinⅡ在肿瘤组织中的平均阳性细胞率（61．34％±11．08％）显著高于周围组织（19．75％±4．59％）（P<0．05）；DTF肿瘤组织中，浸润区平均阳性细胞率（65．22％±14．15％）显著高于非浸润区（46．75±9．01）（P<0．05）。2．DTF肿瘤组织中，TGF-β₁阳性率为79．4％，其中+为29．4％（10／34），++26．5％（9／34），+++23．5％（8／34）；TGF-βRI阳性率82．3％，其中+为29．4％（10／34），++23．5％（8／34），+++29．4％（10／34）；TGF-β₁和TGF-βTI在肿瘤组织和周围组织中的表达水平均有显著性差异（P<0．05）；TGF-β₁和TGF-βRI表达水平之间呈正相关（P<0．05）。3．DTF肿瘤组织中可见α-SMA的表达，阳性率为67．6％，其中+为14．7％（5／34），++44．1％（15／34），+++8．8％（3／34）；肿瘤细胞中TGF-βRI与α-SMA表达水平之间呈正相关（P<0．05）。4．DTF肿瘤细胞和间质中可见fibronectin的表达，阳性率为100％。DTF肿瘤细胞中TGF-βRI与fibronectin表达水平之间呈正相关（P<0．05）。5．VEGF主要表达于DTF肿瘤细胞，血管内皮呈不连续弱阳性着色，阳性率为88．2％。肿瘤组织MVD为9．14±2．08。TGF-β₁和VEGF平均阳性细胞率与MVD之间均呈正相关（P<0．05）。结论：1．AnnexinⅡ在DTF中过度表达，可能参与DTF的发病，推测其通过介导纤溶酶原激活途径涉及肿瘤的浸润性生长。本研究显示DTF中AnnexinⅡ弥漫强阳性表达，为进一步尝试应用血管抑素靶向治疗DTF提供了实验依据。2．TGF-β₁和TGF-βRI在DTF中均过度表达，可能参与DTF的发病，DTF肿瘤细胞中可能存在TGF-β₁自分泌环。3．DTF肿瘤细胞由纤维母细胞和肌纤维母细胞构成。TGF-β₁／TGF-βRI可能参与诱导部分肿瘤细胞的肌纤维母细胞分化、间质构建及肿瘤血管新生。4．过度表达的VEGF可能也参与诱导肿瘤血管新生。结合我们和以往学者的研究结果，推测其肿瘤细胞VEGF表达的上调可能与TGF-β₁／TGF-βRI途径调节有关。更多还原

【Abstract】 BackgroundDesmoid-type fibromatoses, also known as aggressive fibromatosis, are locally aggressive clonal fibroblastic proliferations that arise in deep soft tissues and are characterized by infiltrative growth and a tendency toward local recurrence, but an inability to metastasise. Treatment of patients with DTF usually involves surgery with the goal of complete local excision of the tumor. AnnexinⅡis a well established receptor for plasminogen and tissue plasmin activator, and may be a attractive target for anti-breast cancer therapy since it has been shown to bind to angiostatin. No study about the expression of AnnexinⅡin DTF was found in Pubmed and Wangfang database （1980-2008）. TGF-β₁ is a multifunctional protein which plays an important role in cell proliferation, differentiation and apoptosis. It has been found in our previous study that TGF-β₁ may involve inducing transdifferentiaton of fibroblast to myofibroblast and angiogenesis in nodular fasciitis. Although TGF-β₁ expression in DTF has been demonstrated in recent two papers, its role in DTF is still not well known.Objectives:1. To investegate the expression of AnnexinⅡin DTF and its possible role in DTF.2. To confirm the expression of TGF-β₁ in the DTF, and then to explore its possible role in DTF.Methods:The 34 cases of DTF tumor and matched surrouding tissues were studied byimmunohistochemistry stainings. The expressions of AnnexinⅡ, TGF-β₁/TGF-βRIand VEGF were examined, and the relationship between their expressions andmyofiberblast differentiaton, stroma production and angiogenesis in DTF wasestimated.Results:1. AnnexinⅡwas expressed in 67.6% （29/34） cases of DTF. The expression was markedly in infiltrating region. The average percentage of AnnexinⅡpositive cells （61.34%±11.08%） in DTF was significantly higher than that （19.75%±4.59 %） of surrounding tissues（P<0.05）. Within the positive cases of DTF, the average percentage of AnnexinⅡpositive cells in infiltrating areas （65.22%±14.15%） was significantly higher than that in non-infiltrated ones（46.75±9.01）（P<0.05）. 2. TGF-β₁ was detected in 79.4%（27/ 34） DTF, with a different expressing level [+ in 29.4% （10/34）, ++ in 26.5%（9/34）,and +++ in 23.5% （8/34） respectively]. TGF-βRI was detected in 82.3% DTF[+ in 29.4%（10/34）, ++ in 23.5% （8/34）, and +++ in 29.4% （10/34） respectively]. There was a significant difference between the expression levels of both of TGF-β₁ and TGF-βRI in DTF and those of surrounding tissues （P<0.05） .3. 67.6% （23/34） cases of DTF shownα-SMA positive in the tumor spindle cells [+ in 14.7% （5/34）, ++ in 44.1%（15/34）, and +++ in 38.8%（3/34）]. There was a positive correlation between the expression level ofα-SMA and that of TGF-βRI in DTF （P<0.05）.4. Fibronectin was demonstrated in all cases of DTF. There was a positive correlation between the expression level of fibronectin and that of TGF-βRI in DTF （P<0.05）.5. The MVD in DTF was 9.14±2.08.VEGF positive spindle cells and endothelium were found in 88.2%（30/34）DTF. There was a positive correlation between MVD and the average percentage of both VEGF and TGF-β1 positive cellsConclusion:1. There is a overexpression of AnnexinⅡin DTF. AnnexinⅡmay participated in the development of DTF, possibly in promoting the infiltrative growth of tumour cells by mediating plasminogen activation pathway. AnnexinⅡreactivity may be the basis of a new attractive target for anti-DTF therapy.2. Overexpressions of both TGF-β₁ and TGF-βRI in DTF indicate that TGF-β₁ may participated in the development of DTF. The results in our study suggest it might envolved in neoplastic myofiberblast differentiaton, stroma production and angiogenesis in DTF.3. VEGF overexpression is present in DTF. Diffuse and strong VEGF reactivity in DTF surggests it may be another target for anti-angiogenic and anti-DTF therapies.更多还原