【作者】 宋忠臣；

【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】 口腔颌面部肉瘤（oromaxillofacial sarcoma，OMFS）是一组起源于间叶组织的恶性肿瘤，组织学差异明显，种类繁多，发病率相对较低，但恶性程度多数较高，出现远隔转移早，综合治疗效果差，预后多不良，是临床处理非常棘手的恶性肿瘤，这些都迫使人们研究其发病机制，揭示其生物学特性和探索新的治疗手段。肿瘤分子生物学的研究揭示细胞增殖和凋亡失衡是肿瘤发生发展的重要机制。近年来，对细胞凋亡抑制基因的研究已成为分子肿瘤学的一个热点。Survivin是凋亡抑制蛋白（inhibitorapoptosis ofproteins，IAPs）家族的新成员，其周期性地表达在细胞周期的G₂／M期，抑制半胱氨酸蛋白酶（caspase3、7）的活性而发挥抗凋亡功能，也可以调控细胞周期，参与有丝分裂和细胞增殖。Bcl-2蛋白是凋亡调控蛋白之一，能抑制细胞凋亡，减少细胞对生长因子的需求，延长细胞的生存时间，但不能促进细胞增殖。目前Survivin、Bcl-2在口腔颌面部肉瘤中表达及其与细胞增殖和凋亡关系的研究罕见报道。 目的 本课题通过对Survivin、Bcl-2蛋白在OMFS中的表达及其与细胞增殖和凋亡的关系进行研究，初步探讨它们在OMFS发生发展中的作用，深入了解OMFS的生物学行为，并为OMFS的早期诊断，判断预后和指导临床治疗提供一种新思路。 材料和方法 1．收集口腔颌面部肉瘤标本37例，良性间叶组织肿瘤14例和5例正常肌肉或脂肪组织标本。全部组织均经10％福尔马林固定，石蜡包埋，术前均未行放疗、化疗或其他生物治疗。为日人学2004届硕士毕业沦文日腔{项面部肉瘤卜urvlvin、l弓。I一2表达及其与红11胞增殖和调}‘，关系l沟研究 2.应用免疫组织化学S一P法检测Survivin、Bel一2、PCNA表达情况，应用TuNEL拟、原位检测细胞凋亡情况。 3.应用sPsslo.0统计分析软件，采用、’检验，Fishe:精确概率法、方差分析和t险验进行统计学处理，取。=0.05为显著性检验水准】 结果 1.OMFS中Survivin阳性表达率为81.08%，而良性肿瘤中无Survivin表达，两组l’f，J差异有统计学意义（P<0.05）。在高分化、中分化、低分化OMFS中，Survivin阳性表达率分别为58.33%、80.00%、100%，其中高分化组与低分化组间差异有统计学意义 （P<0.05）。survivin与OMFS肿瘤大小，复发和淋巴结转移等生物学行为无关 （外0.05），在发生远处转移的肉瘤中Survivin蛋白虽明显升高，但差异无统计学意义 （产bo.05）。 2.OMFS中Bcl一2蛋白的阳性表达率为67.57%，良性间叶组织肿瘤中其阳性表达率为28.57%，差异有统计学意义（P<0.05）。Bcl一2蛋白阳性表达率在高分化组、中分化组及低分化组中分别为41.67%、70.00%、86.67%，其中高分化组与低分化组间差异有统计学意义（P<0.05）。经统计学分析显示Bcl一2与OMFS有无复发，淋巴结转移和远处转移间差异均无统计学意义（P>0.05）。 3.OMFS组PCNA一LI明显高于良性肿瘤组（尸<0.05）:PCNA一LI随OMFS组织分化程度降低而升高，在高分化组与中分化组间，高分化组与低分化组间，以及中分化组与低分化组间差异均有统计学意义（P<0.OI）:复发组肉瘤PCNA一LI与原发组肉瘤PcNA一LI间，差异有统计学意义（P<0.05）;发生淋巴结转移或远处转移的肉瘤中PCNA一Ll虽明显升高，但差异无统计学意义（P>0.05）。 4.OMFS组凋亡指数（Al）明显低于良性肿瘤组Al（尸<0.01）;Al随OMFS组织分化程度降低而降低，经统计学分析，在各组间差异均有统计学意义（P<0.ol）;Al与OMFS的复发，淋巴结转移和远处转移无关（P>.05）。 5.OMFS中，Survivin与Bel一2蛋白表达密切相关（P<0.05）。 6.OMFS中，Survivin阳性组PCNA一Ll 39.51士14.63显著高于Survivin阴性组PCNA一Ll阴性组Al22.52士1 1.65（尸<0.05）:Survivin阳性组Al 3.72士2.50显著低于Survivin6 .39士2.42（尸<0.05）。7.OMFS中，Bcl一2蛋白阳性表达组PCNA一LI显著高于Bcl一2阴性表达组（尸<0.05），哪州人学2004)lti硕士毕业论文IJ月空须面部肉瘤Survlv.「1、Bcl一2友达及其与细胞增殖和凋}’.关系的研究而Bcl一2蛋白阳性表达组Al显著低于Bc卜2阴性表达组（P<0.05）。 8.在5例正常肌肉、脂肪组织中均未检测到Survivin、Bcl一2蛋白、PCNA及凋亡细胞。 结论 1.survivin在OMFS组织中特异性地高表达并与肉瘤的分化程度相关，提示 Survivin可能与OMFS的发生发展有关，是OMFS早期而普遍的事件。 2.Bcl一2蛋白过表达与OMFS组织分化程度有关，提示检测Bd一2蛋白表达对判断 口腔领面部肉瘤的恶性程度具有参考价值。 3.PCNA增殖指数是反映OMFS细胞增殖活性和分化水平的一个重要指标。 4.细胞凋亡指数是反映OMFS恶性程度，评价其生物学行为的参数。 5，在OMFS中Survivin和Bel一2蛋白表达密切相关，提示Survivin有可能与Bel一2 共同作用抑制凋亡。 6.OMFS中，Survivin异常表达，伴有增殖指数升高，凋亡指数下降，提示survivin 可能通过抑制细胞凋亡从而促使肿瘤细胞的增殖恶化。 7.Bcl一2蛋白与细胞增殖失控、凋亡抑制关系密切，在OMFS的恶性进展中起重要 作用。更多还原

【Abstract】 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. So it is very difficult in treating the malignant tumor clinically. Molecular biology of tumors revealed that the imbalanced cell proliferation and cell apoptosis is an important mechanism of the tumorigenesis and progression. In recent years, the research of inhibitor apoptosis genes has been a hot topic in molecular oncology. Survivin is a new member of the IAPs(Inhibitor apoptosis of proteins) family. It regularly expresses in G2/M phase and suppresses the activity of caspase-3 and caspase-7 to inhibit cell apoptosis. It is also involved in regulation of proliferation. Bcl-2 protein is the most common antiapoptotic protein, which can inhibit cell apoptosis, decrease the need of growth factor for cell, prolong cell life. But Bcl-2 protein cannot contribute to cell proliferation. At present, little is known about study on the expression of Survivin , Bcl-2 as well as their relationship with proliferation activity and apoptosis in OMFS.Objective This study aimed at investigating the expression of Survivin , Bcl-2 aswell as their relationship with proliferation activity and apoptosis in OMFS in order to evaluate their roles in tumorigenesis and progression of OMFS,and understand the biological character of OMFS, so as to provide a potential new way for early diagnosis, prognosis assessment and indicating theraputics of OMFS.Materials and methods1. 37 OMFS samples, 14 benign mesenchymal tumor samples and 5 normal mesenchymal tissue samples were collected. The tissues were fixed in 10% formalin and embedded in paraffin. No patients had received preoperative chemotherapy, radiation therapy or other biological therapy.2. Immunohistochemical streptavidin peroxidase method was used to detect the expression of Survivin , Bcl-2 , PCNA in all samples. Cell apoptosis was examined by TUNEL method.3. The results were analyzed by SPSS 10.0 software wrap, using Chi-square, Fisher’s exact test, analysis of variance (ANOVA) and t-test. Statistically significant level was considered as "alpha equals 0.05".Results1. The Survivin positive rate in oromaxillofacial sarcomas was 81.08% and none in benign tumors. The result showed a significant difference between the two groups(P<0.05). In highly, moderately and poorly differentiation OMFS groups, the positive rates of Survivin expression were 58.33%, 80.00% and 100.00% respectively, and a statistical significance was only observed for Survivin between the highly and poorly differentiation OMFS groups(P<0.05). The results showed no significant difference between the sarcomas with and without lymph node metastasis or sarcomas with local recurrence(P>0.05). The expression level of Survivin was higher in sarcomas with distant metastasis than those without distant metastasis, but there was no significant difference between the two groups(P>0.05).2. The Bcl-2 positive rate was 67.57% in OMFS group and 28.57% in benign tumor group respectively. The difference between them was significant(P<0.05). In highly, moderately and poorly differentiation OMFS groups, the positive rates of Bcl-2 expressionwere 41.67%, 70.00%, 86.67% respectively. And a statistical significance was only observed for Bcl-2 expression between the highly and poorly differentiation OMFS groups. No correlation was observed between expression of Bcl-2 protein and the biological behaviors of OMFS(P>0.05).3. Compared to benign mesenchymal tumors, PCNA proliferation index(PCNA-LI) was significantly higher in OMFS(P<0.05). The expression level of PCNA-LI tended to increase with the decrease of the degree of the histological differentiation. In highly, moderately and poorly differentiation OMFS groups, there were significant diffe更多还原