【作者】 汪楠；

【导师】 袁苏徐；

【作者基本信息】 苏州大学 ， 肿瘤学， 2013， 硕士

【摘要】 目的：①通过免疫组化S-P法和RT-qPCR方法检测Lumican在人大肠癌中的表达；②研究Lumican与上皮粘附分子E-cadherin、基质降解酶MMP-7表达的相关性，初步探讨Lumican在大肠癌中的表达、临床意义及其与大肠癌发生、发展及浸润、转移的关系。方法：收集64例大肠癌组织及其对应的37例癌旁组织、远离癌的27例正常大肠组织蜡块。①应用免疫组化S-P法检测上述同一组织内Lumican、E-cadherin及MMP-7蛋白表达；②分别提取27例新鲜大肠癌组织及远离肿瘤的正常大肠组织RNA，应用荧光定量聚合酶链反应（RT-qPCR)技术检测Lumican mRNA相对表达量；③结合Lumican表达的临床意义，就其与相应同一组织内E-cadherin、MMP-7表达相关性行统计学分析。统计学分析采用SPSS19.0软件。结果：①免疫组化显示Lumican蛋白阳性染色定位于胞膜，癌细胞过表达，癌旁成纤维细胞亦有表达，正常大肠上皮细胞不表达。64例大肠癌、37例癌旁组织Lumican阳性表达率分别为73.44%（47/64）、67.57%（25/37），差异无统计学意义（P>0.05）。27例正常大肠组织无一例表达Lumican，表达率为0%（0/27），显著低于癌组织、癌旁组织表达，差异有统计学意义（P<0.001）。大肠癌组织Lumican蛋白表达与患者性别、年龄、发病部位无关，而与病理分化程度、Dukes分期、淋巴结是否转移相关；分化越差，Dukes分期越晚，伴淋巴结转移，Lumican表达越高，差异均有统计学意义（P<0.001）。②E-cadherin阳性染色定位在上皮细胞-细胞接触侧细胞膜上；MMP-7主要表达在胞质，部分胞膜染色，间质不染或浅染。大肠癌组织E-cadherin、MMP-7阳性表达率分别为34.38%（22/64）、68.75（44/64），两者表达呈负相关（r=－0.648），有统计学意义（P<0.01）。③Lumican mRNA在大肠癌组织中表达高于远离肿瘤的正常组织表达（P<0.001）。同一大肠癌组织免疫组化检测Lumican蛋白量与RT-qPCR检测Lumican mRNA相对表达量呈正相关（r=0.123），但无统计学意义（P>0.05）。④Lumican表达与E-cadherin表达呈负相关（r=－0.570），Lumican与MMP-7表达呈正相关（r=0.701），均有统计学意义（P<0.01）。结论：①大肠癌组织中Lumican mRNA和蛋白均较正常大肠组织表达升高，大肠癌组织Lumican mRNA升高引起翻译生成的Lumican蛋白表达增多。Lumican蛋白表达与大肠癌组织学分级、Dukes分期及淋巴结是否转移有关。②大肠癌中E-cadherin和MMP-7表达与组织学分级、Dukes分期及淋巴结是否转移密切相关，E-cadherin低表达与MMP-7高表达在大肠癌的侵袭和转移过程中可能有一定的协同作用。③Lumican蛋白表达与E-cadherin呈负相关，而与MMP-7呈正相关。联合检测Lumican、E-cadherin及MMP-7可作为评价大肠癌恶性生物学行为指标。更多还原

【Abstract】 Objective: To quantify the expression of Lumican in the human colorectal carcinomatissue by immunohistochemistry and RT-qPCR, and to study its correlation with theexpression of E-cadherin and MMP-7, in order to investigate the expression of Lumicanand its significance with the progression of colorectal carcinoma.Methods: Protein levels of Lumican, E-cadherin and MMP-7in paraffin sections of64cases of colorectal carcinoma,37cases of adjacent mucosa and27cases of distantnormal mucosa were determined by immunohistochemical Streptavidin-perxiodasetechnique, mRNA levels of Lumican of27cases of colorectal carcinoma and distantnormal mucosa was quantified by RT-qPCR. Additionally, clinical parameters includinggender and age of these patients and the site, grade were recorded, the protein levels ofE-cadherin and MMP-7in the same colorectal carcinoma tissue were determined byimmunohistochemistry. Finally, the relationship between the expression of Lumican ofE-cadherin, MMP-7in protein levels in the same patients with colon carcinoma wereanalyzed by SPSS.Results:①.Immunohistochemical results:Based on its immunoreactivity, theLumican protein was found to be localized in the cell membrane of colorectal cancer cellsand fibroblasts of stromal tissues adjacent to cancer cells, but not in colorectal carcinomaand adjacent normal mucosa were73.44%and67.57%, which were significantly higherthan that in distant normal mucosa of0%(p<0.001). The expression of Lumican incolorectal carcinoma were significantly correlated with the tumor differentiation, theDukes stage and lymph node metastasis, but there is no relationship between the Lumicanexpression and age, sex, attacking location and the size of tumor.②.The expression ofLumican mRNA in colorectal carcinoma were higher than that in distant normal mucosa.The expression of Lumican protein was positively correlated with Lumican mRNA in the same colorectal carcinoma, but this was not statistically significant (p>0.05).③.Based onits immunoreactivity, the E-cadherin protein was found to be localized in the cellmembrane of colorectal cancer cells, MMP-7protein was expressed in both of cellmembrane and cytoplasma, but weekly in the stromal tissues. The positive rate ofE-cadherin and MMP-7protein expression in colorectal carcinoma wererespectively:34.38%%and68.75%. There was a negative association between E-cadherinprotein and MMP-7（r=－0.648, P<0.01）.④.There was a negative association betweenLumican protein and E-cadherin（r=-0.570, P<0.01）, whereas a positive associationbetween Lumican and MMP-7（r=0.701, P<0.01）.Conclusion: The Lumican proteins and mRNA in colorectal carcinoma weresignificantly higher than those in distant normal mucosa. The expression of Lumican incolorectal carcinoma were significantly correlated with the tumor differentiation, theDukes stage and lymph node metastasis. Lower E-cadherin and higher MMP-7expressionmay be positively correlated with the development of the relative later stages of colorectalcarcinoma. The expression of Lumican protein and E-cadherin, MMP-7was correlated incolorectal carcinoma. The above three interacted with each other during the progression ofcolorectal carcinoma.更多还原