【作者】 李杰茹；

【导师】 齐凤英； 韩彩丽；

【作者基本信息】 河北医科大学 ， 病理学与病理生理学， 2004， 硕士

【摘要】 目的：食管癌是我国常见的消化道恶性肿瘤之一，其主要死亡原因是术后转移。大量研究表明，食管癌的形成和转移是一个多基因、多因子共同作用的结果。本研究通过对食管癌组织中MMP-2、TIMP-2、CD34和CD44v6蛋白表达的检测，探讨MMP-2、TIMP-2、CD34和CD44v6与食管癌的发生、发展及转移的关系；MMP-2和TIMP-2蛋白表达及MMP-2和CD44v6蛋白表达与食管癌浸润转移的相关性，MMP-2对食管癌组织血管生成的影响，为预测食管癌浸润转移提供客观参考指标。方法：1 应用免疫组织化学（immunohistochemistry，IHC）方法检测了48例食管癌组织，17例癌旁不典型增生组织和12例切缘正常组织中MMP-2蛋白的表达和平均血管密度（mean microvessel density，MVD）。2 应用流式细胞术（flow cytometry，FCM）的方法定量测定了30例食管癌组织，14例癌旁不典型增生组织和5例切缘正常组织中MMP-2、TIMP-2和CD44v6蛋白的表达。3图像分析：采用HPIAS-1000病理图文分析系统，将IHC染色结果转化为灰度值进行测量。结果： 1 MMP-2在食管上皮不同病变中的表达 <WP=4>采用FCM定量检测30例食管癌组织、14例癌旁不典型增生组织和5例切缘正常组织标本中MMP-2的含量，结果显示三者之间存在着明显的差别（P<0.001），且MMP-2在癌组织中的表达明显高于癌旁组织和切缘正常组织（P<0.05）（表2），说明食管癌组织具有较强分泌MMP-2的能力。应用IHC法对48例食管癌、17例癌旁不典型增生组织和12例切缘正常组织标本中MMP-2蛋白表达的检测获得基本相似的结果（表3，4），所不同之处为，应用IHC法检测时，癌旁组织MMP-2表达显著高于正常组织（P<0.05）（表3，4）。而应用FCM法检测时，MMP-2在癌旁组织中的表达与正常组织间无明显的差异（P>0.05）（表2）。综合IHC与FCM检测结果，MMP-2表达与患者的年龄、性别及肿瘤的分化程度无关（P>0.05），但MMP-2蛋白的表达与淋巴结转移呈正相关（P<0.05），且MMP-2蛋白的表达与浸润深度呈正相关（P<0.05）（表1，6，7）。2 TIMP-2在食管上皮不同病变中的表达采用FCM检测30例食管癌组织、14例癌旁不典型增生组织和5例切缘正常组织标本中TIMP-2的表达发现，三者之间存在着差别，但此差别无显著性意义（P>0.05）（表5），TIMP-2的表达与患者的性别、年龄、浸润深度及分化程度均未见明显的相关性（P均>0.05），而与淋巴结转移密切相关（P<0.05）（表8）。3食管上皮不同病变中的MMP-2/TIMP-2计算MMP-2 tumor/normal/TIMP-2 tumor/normal间的比值，这一比值反应了二者之间的平衡。发现FCM检测30例食管癌组织、14例癌旁不典型增生组织和5例切缘正常组织<WP=5>标本中，MMP-2 tumor/normal/TIMP-2 tumor/normal间的比值有明显的不同(P <0.001)，癌组织中比值明显高于癌旁不典型增生组织和切缘正常组织(P <0.05)（表9）。18例有淋巴结转移的患者中，比值为1.7879±0.4003，12例无淋巴结转移的患者中，比值为0.9725±0.1675，MMP-2/TIMP-2间的平衡与有无淋巴结转移密切相关（P <0.005）,并且MMP-2/TIMP-2间的平衡与肿瘤的浸润深度有关(P<0.05)；MMP-2/TIMP-2间的平衡与患者的性别、年龄、分化程度均未见明显的相关性（P均>0.05）（表11）。4在食管上皮?更多还原

【Abstract】 Objective: Esophageal carcinoma is one of the high incidence carcinomas in malignant digestive tumors in our country. Patients with esophageal carcinoma were died from metastasis of esophageal carcinoma. It has been known that combined biological effects of multiple genes and multiple factors are involved in the carcinogenesis of esophageal epithelium and metastasis of esophageal carcinoma. To investi- gate the expression of matrix metalloproteinases-2(MMP-2), Tissue inhibitor of metalloproteinase-2 (TIMP-2), CD44v6 protein and intratumor microvessel density(MVD) in esophageal carcinoma and the relationships to biological behavior of the cancer. To investigate relationships between expression of MMP-2 and TIMP-2 and their correlation to invasion and metastasis in esophageal cancer. To investigate relationships between expression of MMP-2 and CD44v6 and their correlation to invasion and metastasis in esophageal cancer. To investigate the effect of MMP-2 on angiogenesis . The results may be helpful in prognostication of esophageal cancer.Methods:1 The expression of MMP-2 protein and MVD were examined by s-p immunohistochemical stain (IHC) in 48 esophageal <WP=9>carcinoma specimens and 17 para-tumorous esophageal tissues and 12 incisal margin normal tissues.2 The expression of MMP-2 protein, TIMP-2 protein and CD44v6 protein in 30 esophageal carcinoma specimens and 14 para-tumorous esophageal tissues and 5 incisal margin normal tissues were quantitatively examined by flow cytometry(FCM).3 Image Analysis: The results of IHC is applied to HPIAS-1000 image analysis system and translated to gray values.Results:1 Expression of MMP-2 in various lesions of esophageal epithelialThere was a significant difference in 30 esophageal carcinoma specimens and 14 para-tumorous esophageal tissues and 5 incisal margin normal tissues were quantitatively examined by FCM(P<0.05) (table 2), and expression of MMP-2 in esophageal carcinoma was significantly higher than in para-tumorous esophageal tissues and incisal margin normal tissues(P<0.05). It showed that esophageal carcinoma cell had a strong capability to secrete MMP-2. Expresions of MMP-2 protein in 48 esophageal carcinoma specimens and 17 para-tumorous esophageal tissues and 12 incisal margin normal tissues were detected by IHC, the results was similar to that of MMP-2 protein detected by FCM(table 3,4). But there was a difference between the result of IHC and the result of FCM, Expression of MMP-2 in para-tumorous esophageal tissues was <WP=10>significantly higher than in incisal margin normal tissues(P<0.05), but there wasn’t a significant difference in para-tumorous esophageal tissues and incisal margin normal tissues were quantitatively examined by FCM (P>0.05).Results of IHC and FCM showed that expression of MMP-2 wasn’t correlated with age, sex and grate of differentiation (P>0.05), but the high expression of MMP-2 in cancer tissues were significantly related with the fibromembranous invasion and lymph node metastasis (P<0.05)( table 1,6,7).2 Expression of TIMP-2 in various lesions of esophageal epithelialThere wasn’t a significant difference in 30 esophageal carcinoma specimens and 14 para-tumorous esophageal tissues and 5 incisal margin normal tissues were quantitatively examined by FCM (P>0.05)( table 5). Expression of TIMP-2 wasn’t correlated with age, sex , grate of differentation and fibromembranous infiltration (P>0.05), but the low expression of TIMP-2 in cancer tissues was significantly related with lymph node metastasis(P<0.05) (table 8).3 MMP-2/TIMP-2 in various lesions of esophageal epithelialStudies in MMP-2(tumor/normal)/TIMP-2(tumor/normal) could indicate the balance between MMP-2 and TIMP-2. There was a significant difference in 30 esophageal carcinoma specimens and 14 para-tumorous esophageal tissues and 5 incisal margin normal tissues were quantitatively examined by <WP=11>FCM(P<0.001),and expression of MMP-2/TIMP-2 in esophageal carcinoma was significantly higher than in para-tumorous esophageal tissues and更多还原