【作者】 张斌；

【导师】 僧国珍；

【作者基本信息】 郑州大学 ， 普通外科， 2004， 硕士

【摘要】 [背景和目的]：肿瘤的主要特征是无控制侵袭性生长及转移。胃癌是常见的消化道肿瘤之一，目前胃癌的发病率，死亡率仍居第一位。在消化系统恶性肿瘤的死亡病例中，约有半数死于胃癌，目前其治疗方法包括：手术、化疗、放疗、生物治疗，免疫治疗等综合疗法。然而其治愈率并未明显提高，而癌转移是造成胃癌死亡的主要原因。目前癌转移的机制尚不清楚，因此，如何诊断及阻断肿瘤的浸润转移引起了广泛的关注。细胞外基质(Extracellular matrix，ECM)和基底膜是机体对抗恶性肿瘤转移的第一道屏障，肿瘤浸润转移的过程需要细胞外基质(ECM)的降解，基质金属蛋白酶(matrix metalloproteinases，MMPs)在ECM降解中起重要作用，基质金属蛋白酶(MMPs)是一组重要的锌依赖蛋白酶，它主要参与细胞外基质的降解与转运。MMP-7属于MMPs基因家族成员之一，其作用尤为重要，MMP-7主要表达与肿瘤细胞中，MMP-7能降解Ⅳ型胶原，纤维连结蛋白和层粘连蛋白，纤维结合素，蛋白多糖，Ⅰ型明胶和可溶性弹性蛋白等底物。肿瘤细胞通过分泌MMP-7来降解基底膜的ECM成分，从而破坏了机体防御肿瘤浸润与转移的自然屏障，增加了癌细胞的侵袭性，促进了肿瘤的浸润和转移。随着分子生物学技术的发展，对胃癌浸润转移的分子学基础有了较深入的研究。核转录因子KAPPAB(nuclear factor kappaB，NF-κB)成为近年研究的热点。郑州大学2(X)4年硕士论文NF。K B andMMP一7在胃癌中的表达及临床病理意义NF一忍广泛存在于真核生物中，是一种重要的核转录因子，有一个复杂的体系组成。它能促进多种细胞因子的表达，它作为信号传导途径中的枢纽与肿瘤的发生发展，细胞凋亡的调节以及胚胎发育等重要事件有着密切关系，有报道称它对MMP一7有重要的调节作用，通过促进MMP一7的表达来增强肿瘤的浸润转移能力;本实验通过对胃癌中NF一虑、MMP一7的表达进行研究，初步探讨它们在胃癌浸润转移中的作用，二者之间的相互关系及可能的分子机制。防法】:采用免疫组化PV-9000法检测NF一沼p65、MMP一7蛋白在48例胃癌组织中，15例正常组织中，15例肠上皮化生，18例不典型增生组织中的表达;探讨它们在胃癌浸润转移中的作用，二者之间的相互关系及可能的分子机制。分析它们在不同病变中的表达与胃癌临床病理参数的关系。统计分析采用SPSS 10.0软件包进行统计分析。采用扩检验，Fisher精确概率法及spe~an等级相关对相应资料进行分析，取a=0.05为检验水准。【结果卜1.NF一妞p65表达情况:(l)在48例胃癌标本中33例NF一沼p65表达 (33/48，·68.8%)明显高于正常胃组织，肠上皮化生组织及胃不典型增生组织中的表达，差异具有显著性:(2)在22例淋巴结转移的肿瘤组织中20例NF一虑p65表达阳性(20/22，90.9%)，而在26例无淋巴结转移的肿瘤组织中仅有13例阳性(13/26，50.0%)，差异具有显著性(尸<0.05);浸润至浆膜外组表达率88.9%(8/9)明显高于侵及浅肌层组38.9%(7/18)，差异具有显著性(尸<0.05);PTNM分期111一W患者阳性率94.7%(18/19)明显高于I一H患者阳性率51.7%(15/29)，差异具有显著性;(3)NF一沼p65的表达还与胃癌分化程度，肿瘤大小密切相关，与年龄、性别、肿瘤部位等因素无关。 2.MMP一7表达情况:(l)在48例胃癌标本中29例MMP一7表达阳性，阳性率为60.4%(2 9/48)，明显高于正常组织，肠上皮化生组织及不典型增生中的表达，差异具有显著性;(2)在22例淋巴结转移的肿瘤组织中19例MMP一7表达(19/22，86.4%)。而在26例无淋巴结转移的肿瘤组织中仅有10例阳性表郑州大学2004年硕士论文NF。K B andM州lP一在胃癌中的表达及临床病理意义达(10/26，36.5%)，差异具有显著性(尸<0 .05);侵及浆膜(15/21)，浆膜外层组(8/9)的表达明显高于浸及浅肌层组(6/18)，差异具有显著性;MMP一7在肿瘤大于5 cM组的阳性率86.9%(20/23)明显高于小于5 cM组的阳性率36.0%(9/25)，差异具有显著性;PTNM分期m一W患者阳性率94.7%(18/19)明显高于I一H患者阳性率37.9%(11/29)，差异具有显著性;MMP一7的表达还与胃癌分化程度密切相关，与年龄、性别、肿瘤部位等因素无关。 3.sP~an相关分析表明NF-虑p65的表达与MMp一7的表达成正相关。rs=0 .779，(P<0.01).I结论l:1.NF一忍在胃癌中的表达显著高于相应正常组织，并且与胃癌的浸润深度、淋 巴结转移、肿块大小及PTNM病理分期密切相关，提示NF一虑在胃癌发展浸 润转移中起重要促进作用。2.MMP一7的表达显著高于相应正常组织，并且与胃癌的浸润深度、淋巴结转 移、FINM病理分期及肿块大小密切相关，提示MMP一7在胃癌发展浸润转 移中起重要促进作用。3.胃癌中NF一沼在蛋白水平上对MMP一7有重要的调节作用，通过促进MMP一7 的表达来促进肿瘤的浸润转移。4.胃癌中NF一沼、MMP一7共阳性表达与胃癌的浸润深度、淋巴结转移及PTNM 病理分期成正相关。联合检测NF一忍、MMP一7的表达，可以作为病情监控指 标，可能有利于估计胃癌患者的预后。同时以NF一虑、MMP一7为中心的靶向 治疗，可能成为抑制肿瘤转移的有潜力的途径之一。5.NF*虑、更多还原

【Abstract】 Background and Objective: Gastric carcinoma is one of the most common malignant neoplasms in the world,it is the first frenquent cause of cancer death in our country. There are some treatments to gastric carcinoma,including surgery, chemotherapy, radiotherapy, biotherapy and immunotherapy. But the effection is still poor. It is known that the important dead reason of gastric carcinoma depends on its invasive growth and metastasis. But the molecular mechanism of cancer metastasis is not clear. So how to diagnose malignant neoplasm and prevent invasive growth and metastasis of cancer is important. Extracellular matrix is the first barrier to suppress the invasion and metastasis of cancer. During the invasive growth and metastasis of cancer,it needs degrade Extracellular matrix(ECM). Matrix metalloproteinases(MMPs) play an important role in degrading ECM. MMP-7 is one of the members of MMPs family. It is mainly produced from tumor cells,and has a great role in tumor invasion and metastasis. It can degrade type IV collagen,laminin,fibronectin, proteinpolysaccharide and others. It is related to degrade and remodel the ECM. The carcinoma can destroy the resistence of the body by secreting MMP-7. NF-kB is animportant nuclear factor. It can induce the expression of some cytokines. It was proved that NF-kB play an important role in the activation of immune cell, anti-virus,various stress action, regulation of apoptosis and the occurrence of carcinoma. It is reported that NF-kB can induce the expression of MMP-7 and strengthen the ability of malignant tumor’s invasion and matastasis. By observing the expression of NF-kB,MMP-7 in gastric carcinoma. We try to explore the influence on tumor invasion and matastasis,their mutual relationship and their possible molecular mechanisms.Methods: In this study, NF-kB and MMP-7 were detected by use of immunohistochemical assay from 48 gastric carcinoma specimens and 15 normal gastric mucosa, 15 intestinal metaplasia and 18 gastric hyperplasia mucosa.We observe their expression to explore the influence on tumor invasion and matastasis their mutual relationship and their possible molecular mechanisms.We analysis the relationship between the expression of NF-kB and MMP-7 and the clinicopathologic parameters of gastric carcinoma. Data were performed with SPSS 10.0 software. Chi-square test fisher’ exact test of probabilities and Spearman rank correlation were used, and Statistically significant level was considered as "alpha equals 0.05". Results: 1. The expression of NF-kB :(1) The expression of NF-kB was detected in 33 of 48 gastric carcinoma (68.8%,33/48) , was significantly higher than those in normal gastric mucosa, intestinal metaplasia and gastric hyperplasia mucosa.(2). The expression of NF-kB was detected in 20 of 22 gastric carcinoma with lymph node metastasis (90.9%,20/22) ;But 13 in 26 gastric carcinoma without lymph node metastasis(50.0 %,13/26) ,The difference was significant (P<0.05); The expression of NF-kB in invasion to chorion(15/21 )was significantly higher than those invasion to low muscular layer (6/18); According to clinical stage of PTNM, The differencebetween III -IV (94.7%, 18/19 ) and I - II (51.7%, 15/29) was significant.(3).The expression of NF-kB has significant correlation to tumor size and differentiation.2. The expression of MMP-7: (l).The expression of MMP-7 was detected in 29 of 48 gastric carcinoma(60.4%,29/48), was significantly higher than those in normal gastric mucosa, intestinal metaplasia and gastric hyperplasia mucosa.(2). The expression of MMP-7 was detected in 19 of 22 gastric carcinoma with lymph node metastasis(86.4%,19/22);But 10 in 26 gastric carcinoma without lymph node metastasis (36.5%,10/26),The difference was significant (P<0.05); The expression of MMP-7 in invasion to chorion (15/21) was significantly higher than those invasion to low muscular layer(6/18);The expression of MMP-7 in tumor size 5CM (87.0%,20/23) was significantly higher than those size<5CM (36.0%,9/25); The difference between III-IV(94.7%, 18/19) and I-II更多还原