E-钙黏附素基因甲基化及蛋白在食管鳞状细胞癌中的表达及其临床意义研究
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摘要
目的：
通过比较食管鳞状细胞癌及其相应癌旁组织中E-cadherin基因甲基化和蛋白表达水平差异，探讨其与临床病理资料关系。方法：
收集南昌大学第一附属医院心胸外科行手术治疗的食管鳞癌患者标本50例，取材为食管癌组织及相应的癌旁组织，提取癌组织及相应癌旁组织的基因组DNA，应用甲基化特异性PCR对所提取的DNA进行E-cadherin基因甲基化检测；采用免疫组化SP法对癌组织及癌旁组织中上述基因蛋白表达水平进行检测，并结合临床病理资料进行相关性分析。患者对该实验知情同意，并经医院伦理委员会批准。
结果：
1.食管鳞癌组织中E-cadherin基因启动子甲基化的阳性率为58％（29/50），相应的癌旁食管黏膜组织E-cadherin基因启动子甲基化的发生率为30%（15/50）。癌旁食管黏膜组织E-cadherin基因启动子甲基化的阳性率显著低于癌组织（X2=7.955，P=0.005）。本实验中该基因甲基化发生率在年龄、性别、肿瘤大小和分化程度等各组间均无统计学差异（P＞0.05），在肿瘤的分期各组间及有无淋巴结转移组间有显著差异性，具有统计学意义（P＜0.05）。
2.食管鳞癌组织中E-cadherin主要定位于细胞膜/细胞浆上，E-cadherin蛋白表达的阳性率为40%(20/50)，E-cadherin蛋白表达在患者的年龄、性别、肿瘤大小和分化程度各组间无统计学意义（P＞0.05），在肿瘤的分期各组间及有无淋巴结转移组间有显著性差异（P＜0.05），具有统计学意义。
3. E-cadherin基因的启动子甲基化和相应蛋白的失表达密切相关（P=0.001，r=-0.463）。
结论：
1.食管鳞状细胞癌中E-cadherin基因启动子甲基化阳性率高于正常食管粘膜组织，E-cadherin基因启动子甲基化可能参与了食管鳞状细胞癌的恶性病变过程。
2.食管鳞状细胞中E-钙黏附素呈低表达或不表达,且与肿瘤临床病理分期及淋巴结转移有关，提示细胞间黏附作用的丧失参与了肿瘤的发生发展过程。
3.食管鳞状细胞癌中E-cadherin基因启动子甲基化与相应蛋白质下调或缺失相关。
Objective:
To detect the E-cadherin gene methylation and protein expression in esophagealsquamous cell carcinoma and corresponding adjacent tissue, explore its relationshipwith clinicopathological data.
Methods:
Collect the clinical data of50patients with esophageal squamous cell carcinomain the Department of Cardiothoracic Surgery, The First Affiliated Hospital ofNanchang University. Drawn parts of the tumor tissue and corresponding adjacenttissues (normal esophageal mucosa from the tumor>5cm).The genomic DNA fromthese patients was obtained from the frozen tumor specimens and matched normalmucosal tissue specimens using DNA extraction kit. Methylation changed in thepromoter region of E-cadherin gene was determined by using methylation-specificPCR.Immunohistochemical staining of E-cadherin was performed in the tumortissues using SP two-step method. And combined with clinical data to analysis.Informed consent was obtained from each patients.This experiment was approved bythe hospital ethical committee.
Results:
1. The positive rates of genes promoter hypermethylation of E-cadherin was58%(29/50) in tumor tissue, which was30%(15/50) in normal mucosal tissue. ThePositive rate was significantly higher in tumor tissues than in normal mucosal tissue（X2=7.955，P=0.005）. The methylation frequency of E-cadherin in these specimensdid not differ according to the clinical features: age, gender, tumor size anddifferentiation. And it associated with tumor stage and lymph node metastasis
2. E-cadherin is mainly expressed on the cell membrane/Cytoplasm in the ESCC,the positive rates of protein expression of E-cadherin was40%(20/50), E-cadherinexpressed in these specimens did not differ according to the clinical features: age,gender, tumor size and differentiation. And it associated with tumor stage and lymphnode metastasis
3.The promoter hypermethylation of E-cadherin gene was remarkably associatedwith the loss of protein (P=0.001，r=-0.463).
Conclusion:
1. Promoter hypermethylation of E-cadherin is common in ESCC and occursmore frequent in tumor tissues than in normal mucosal tissues. E-cadherin genepromoter methylation may be involved in the malignant lesions process of ESCC.
2. The methylation E-cadherin is associated with loss of protein，the promptedintercellular adhesion loss involved in the process of tumor development.
3.The promoter hypermethylation of E-cadherin gene is remarkably associatedwith the loss of protein.
通过比较食管鳞状细胞癌及其相应癌旁组织中E-cadherin基因甲基化和蛋白表达水平差异，探讨其与临床病理资料关系。方法：
收集南昌大学第一附属医院心胸外科行手术治疗的食管鳞癌患者标本50例，取材为食管癌组织及相应的癌旁组织，提取癌组织及相应癌旁组织的基因组DNA，应用甲基化特异性PCR对所提取的DNA进行E-cadherin基因甲基化检测；采用免疫组化SP法对癌组织及癌旁组织中上述基因蛋白表达水平进行检测，并结合临床病理资料进行相关性分析。患者对该实验知情同意，并经医院伦理委员会批准。
结果：
1.食管鳞癌组织中E-cadherin基因启动子甲基化的阳性率为58％（29/50），相应的癌旁食管黏膜组织E-cadherin基因启动子甲基化的发生率为30%（15/50）。癌旁食管黏膜组织E-cadherin基因启动子甲基化的阳性率显著低于癌组织（X2=7.955，P=0.005）。本实验中该基因甲基化发生率在年龄、性别、肿瘤大小和分化程度等各组间均无统计学差异（P＞0.05），在肿瘤的分期各组间及有无淋巴结转移组间有显著差异性，具有统计学意义（P＜0.05）。
2.食管鳞癌组织中E-cadherin主要定位于细胞膜/细胞浆上，E-cadherin蛋白表达的阳性率为40%(20/50)，E-cadherin蛋白表达在患者的年龄、性别、肿瘤大小和分化程度各组间无统计学意义（P＞0.05），在肿瘤的分期各组间及有无淋巴结转移组间有显著性差异（P＜0.05），具有统计学意义。
3. E-cadherin基因的启动子甲基化和相应蛋白的失表达密切相关（P=0.001，r=-0.463）。
结论：
1.食管鳞状细胞癌中E-cadherin基因启动子甲基化阳性率高于正常食管粘膜组织，E-cadherin基因启动子甲基化可能参与了食管鳞状细胞癌的恶性病变过程。
2.食管鳞状细胞中E-钙黏附素呈低表达或不表达,且与肿瘤临床病理分期及淋巴结转移有关，提示细胞间黏附作用的丧失参与了肿瘤的发生发展过程。
3.食管鳞状细胞癌中E-cadherin基因启动子甲基化与相应蛋白质下调或缺失相关。
Objective:
To detect the E-cadherin gene methylation and protein expression in esophagealsquamous cell carcinoma and corresponding adjacent tissue, explore its relationshipwith clinicopathological data.
Methods:
Collect the clinical data of50patients with esophageal squamous cell carcinomain the Department of Cardiothoracic Surgery, The First Affiliated Hospital ofNanchang University. Drawn parts of the tumor tissue and corresponding adjacenttissues (normal esophageal mucosa from the tumor>5cm).The genomic DNA fromthese patients was obtained from the frozen tumor specimens and matched normalmucosal tissue specimens using DNA extraction kit. Methylation changed in thepromoter region of E-cadherin gene was determined by using methylation-specificPCR.Immunohistochemical staining of E-cadherin was performed in the tumortissues using SP two-step method. And combined with clinical data to analysis.Informed consent was obtained from each patients.This experiment was approved bythe hospital ethical committee.
Results:
1. The positive rates of genes promoter hypermethylation of E-cadherin was58%(29/50) in tumor tissue, which was30%(15/50) in normal mucosal tissue. ThePositive rate was significantly higher in tumor tissues than in normal mucosal tissue（X2=7.955，P=0.005）. The methylation frequency of E-cadherin in these specimensdid not differ according to the clinical features: age, gender, tumor size anddifferentiation. And it associated with tumor stage and lymph node metastasis
2. E-cadherin is mainly expressed on the cell membrane/Cytoplasm in the ESCC,the positive rates of protein expression of E-cadherin was40%(20/50), E-cadherinexpressed in these specimens did not differ according to the clinical features: age,gender, tumor size and differentiation. And it associated with tumor stage and lymphnode metastasis
3.The promoter hypermethylation of E-cadherin gene was remarkably associatedwith the loss of protein (P=0.001，r=-0.463).
Conclusion:
1. Promoter hypermethylation of E-cadherin is common in ESCC and occursmore frequent in tumor tissues than in normal mucosal tissues. E-cadherin genepromoter methylation may be involved in the malignant lesions process of ESCC.
2. The methylation E-cadherin is associated with loss of protein，the promptedintercellular adhesion loss involved in the process of tumor development.
3.The promoter hypermethylation of E-cadherin gene is remarkably associatedwith the loss of protein.
引文
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