【作者】 谢卫红；

【导师】 李新明；

【作者基本信息】 郑州大学 ， 口腔颌面外科学， 2004， 硕士

【摘要】 背景和目的：口腔颌面部肉瘤(Oromaxillofacial sarcoma，OMFS)的恶性度较高，发展较快，且综合治疗效果差，预后不良，早期诊断和根治性外科手术是提高生存率的主要途径。因此充分利用现代免疫学和分子生物学理论和技术，探索OMFS的发生发展规律，揭示其生物学特性，为临床诊治提供可靠依据尤为必要。粘着斑激酶(focaladhesion kinase，FAK)是一个胞浆酪氨酸激酶，能激活Src内源性蛋白酪氨酸激酶。癌基因、细胞外基质(extracellular matrix，ECM)、整合素(integrin)刺激后，FAK发生酪氨酸磷酸化。FAK的高表达和酪氨酸磷酸化与肿瘤的生成能力相关。大量文献报道，FAK的高表达与多种恶性肿瘤的侵袭和转移正相关。基质金属蛋白酶-9(matrixmetalloproteinase-9，MMP-9)属于明胶酶类，主要作用是降解ECM，其作用底物主要是明胶和Ⅳ／Ⅴ型胶原，同时在血管生成方面发挥作用，促进肿瘤的侵袭和转移。以往大量的研究表明，MMP-9可在皮肤癌、甲状腺癌、前列腺癌、结肠癌、肺癌、骨肉瘤等肿瘤组织中表达，免疫组化研究表明MMP-9表达部位主要在细胞浆，尤其表达在肿瘤侵袭和转移前沿；而且，MMP-9的表达与多种肿瘤的病理分级正相关，MMP-9的表达可能成为恶性肿瘤预后的一项指标。尿激酶型纤溶酶原激活物(urokinase-typeplasminogen activator，uPA)通过激活纤溶酶原增加纤溶酶的生成，从而促进基底膜和细胞外基质蛋白的降解，影响细胞的迁徙。uPA和其受体的结合启动了蛋白溶解过程，有利于MMP-9的激活，促进ECM的溶解。这些过程在肿瘤细胞的侵袭和转移中发挥重要作用。本课题应用免疫组化方法检测OMFS中FAK、MMP-9和uPA的表郑州大学20(抖年硕士毕业论文口腔领面部肉瘤中FAK、MMP一9和证叭的表达及其生物学意义达，探讨FAK激活及其信号传导在肉瘤发生发展中的作用及其相互关系，为临床评估OMFS复发、侵袭和转移潜能，进行预后监测提供参考依据。 材料和方法: 1.收集1993年2月至2002年5月郑州大学第一附属医院手术切除并经病理证实口腔领面部间叶性肿瘤51例。其中OMI.S37例，所有病例术前均未行化疗、放疗或生物治疗。全部组织均经中性福尔马林液固定，常规脱水后石蜡包埋。口腔正常间叶组织5例，取自口腔领面部外伤手术病人。 2.应用免疫组化S一P法检测OMF.S中FAK.、MMP一9和uPA的表达。 3.应用SPSslo.0统计软件对FAK、M血于一9和uPA的表达与肉瘤患者临床病理特征的关系，以及指标间的相关性进行统计分析，检验结果均以Q=0 .05作为有统计学意义的判断水准。 结果: 1 .FAK阳性信号位于肿瘤细胞胞膜或胞浆内，为弥散性或细颗粒状棕黄色，阳性表达细胞为非均质性，口腔领面部肉瘤中FAK的阳性表达率为62.16%，而良性间叶组织肿瘤中阳性表达率为14.29%，两组比较差异有统计学意义(尸<0.05)。在高分化组、中分化组和低分化组中，FAK的阳性表达率分别为33 .33%、70.00%和80.00%，统计分析表明，高分化组与低分化组之间差异有统计学意义(尸<0.仍)。发生复发、淋巴结转移或远处转移的肉瘤中FAK表达明显升高，但差异不具有统计学意义(尸>0 .05)。与无转移肉瘤相比，转移组FAK表达差异有统寸学意义(尸<0.05)。 2.MMP一9的阳性信号位于肿瘤细胞胞膜或胞浆内，为棕黄色或黄色细颗粒状，阳性表达细胞为非均质性。口腔领面部肉瘤中M血IP一9的阳性表达率为62.16%，明显高于良性间叶组织肿瘤(尸<0.05)，并随肉瘤分化程度的降低而呈升高趋势，统计分析表明，高分化组与低分化组之间差异有统计学意义(尸<0.05)。发生复发、淋巴结转移或远处转移的肉瘤中M入IP.9表达明显升高，但差异不具有统计学意义(尸>0.05)。与无转移肉瘤相比，转移组MMP一9表达差异有统计学意义(尸<0.05)。 3 .uPA的阳性信号为棕黄色细颗粒状，位于肿瘤细胞胞膜或胞浆内，阳性表达细胞为非均质性。口腔领面部肉瘤中uPA的阳性表达率为70.27%，而良性间叶组织肿瘤中阳性表达率为21.43%，两组比较差异有统计学意义(尸<0.05)。在高分化组、中分化组和低分化组中，uPA的阳性表达率分别为41.67%、·70.00%和93.33%，统计分析郑州大学2004举硕士毕业论文口腔领面部肉瘤中I叭K、M劫P一9和ul叭的表达及其生物学意义表明，高分化组和中分化组、中分化组和低分化组之间差异不具有统计学意义(尸>0.05)，而高分化组与低分化组之间差异有统计学意义(尸<0.05)。复发肉瘤中uPA的阳性表达率明显高于原发肉瘤(尸<0.05)。在发生淋巴结转移或远处转移的肉瘤中uPA表达有明显升高趋势，但差异不具有统计学意义(尸>0.05)。 4.口腔领面部肉瘤中，FAK的阳性表达和MN[P一9的阳性表达呈显著相关(尸<0.05);FAK的阳性表达和uPA的阳性表达呈显著相关(尸<0.05);MMP一9的阳性表达和uPA的阳性表达呈显著相关(P<0 .05)。 5.FAK、Ml田一9和uPA在正常口腔肌肉、脂肪组织中均不表达。 结论: 1 .FAK在口腔领面部肉瘤发生和发展过程中起重要作用;FAK的高表达可能成为评估口腔领面部肉瘤临床侵袭和转移潜能的有用指标。 2.MMP一9的高表达是反映口腔领面部肉瘤恶性程度评估更多还原

【Abstract】 Background and Objective: Oromaxillofacial sarcoma(OMFS) is a malignant tumor originated from mesenchymal tissue of oral and maxillofacial region which not only has obvious difference of histology, many kinds and lower incidence rate but also is characterized by high malignant degree, early distant metastasis, insignificant effect for combined therapy and poor prognosis.It is essential for correct treatment decisions and better survival rates to make extensive research on the development mechanisms and the biological character of oromaxillofacial sarcoma, understanding the molecular mechanisms of metastasis is critical with respect to oral tumorigenesis. focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that plays a pivotal role in signal transduction at integrin-linked cellular adhesions, which mediate cell contact with the extracellular matrix. It has been shown to play a role in the survival of anchorage-dependent cells and to be essential for integrin-linked cell migration - processes that are likely to play important roles in the development of malignancies. FAK is an intracellular tyrosine kinase associated with the regulation of cell growth, migration, and survival. The purpose of the current study was to determine whether elevated FAK expression in oromaxillofacial sarcoma was associated with increased invasiveness and metastasis. To detect the expression of FAK and MMP-9 and uPA in oromaxillofacial sarcoma and to study their correlation with clinicopathological data including tumor type, grade and lymph node status.Material and Method:1. Thirty-seven cases of OMFS samples, 14 cases of benign mesenchymal tumors and 5 cases of normal mesenchymal tissues of oral and maxillofacial region were collected at the first teaching hospital of zhengzhou university, patients hadn’t received preoperative chemotherapy or radiation therapy.the tissues were fixed in 10% formalin and embedded in paraffin.2. Immunohistochemical streptavidin peroxidase method was used to detect the expression of FAK and MMP-9 and uPA in OMFS and benign mesenchymal tumors.3. The results were analyzed by SPSS 10.0 software wrap, and a P value of less than 0.05 was considered to be statistically significant.Results1 . In oromaxillofacial sarcomas,positive staining for FAK was mainly observed in the cytoplasm and/or on the membrane of tumour cells as well as some mesenchymal cells, and its positive rate was 62.16% (23/37);FAK express was detected in 14.29% (2/14) of benign tumors,and a significant diffenence was found between the two groups (P <0.05) .in the there groups of highly, moderately and poorly differentiated of OMFS,the positive rates of FAK were 33.33% (4/12), 70.00% (7/10) and 80.00% (12/15) .respectively .There was a significant difference between highly differentiated group and poorly differentiated group(P <0.05) .the expression level of FAK was significantly higher in locally recurrent sarcomas than that in primary sarcomas, while no statistical significance was found (P >0.05) . There was a significant difference between sarcomas with distant metastasis and lymph node metastasis and the other sarcomas (P <0.05 ) .2. In OMFS ,the positive rate of uPA is 70.27 % (26/37), while the positive rate is 21.43 % (3/14) in benign tumors, There was a significant difference between the two groups(P <0.05 ) .in the there groups of highly, moderately and poorly differentiated of OMFS,the positive rates of uPA were 41.67% (5/12) and 70.00%(7/10) and 93.33%(14/15), respectively. There was a significant difference between highly differentiated group and poorly differentiated group (P <0.05 ) . the expression level of uPA was significantly higher in locally recurrent sarcomas than in primary sarcomas (P <0.05) .in sarcomas with distantmetastasis and/or lymph node metastasis,the positive expression of uPA tended to increase compared to the other sarcomas, while no statistical significance was found (P >0.05 ) .3. In OMFS , the positive rate of MMP-9 is 62. 16% (23/37);while the positive rate is 21.更多还原