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目的:观察Barrett’s食管黏膜组织中CDX mRNA和蛋白表达,观察不同胆汁酸对人食管细胞株Eca-109CDX2 mRNA表达水平的影响.方法:收集内镜下正常食管组织(n=5),反流性食管炎(n=7),Barrett’s食管(n=8)标本.采用RT-PCR测定CDX1和CDX2 mRNA表达;实时荧光定量PCR测定CDX2 mRNA表达.用免疫组织化学染色法对CDX2蛋白进行定位.人食管癌细胞株Eca-109分别在100、400、1000μmol/L胆酸(CA),脱氧胆酸(DCA),甘胆酸(GC)中培养,分别在0-24h收取细胞,荧光定量PCR测定CDX2 mRNA表达.结果:CDX2 mRNA在正常食管组织中无表达,在Barrett’s食管组、反流性食管炎组表达增高(9.6411±6.6823,3.9156±3.6700),与正常食管组之间比较均存在显著性差异(P=0.006,0.025),但2组之间没有显著性差异(P=0.133),CDX1在所有食管组织中均无表达.免疫组织化学染色在Barrett’s食管有CDX2蛋白胞核呈特异性染色阳性反应,反流性食管炎组织CDX2胞质显示细颗粒性染色(假阳性),正常的食管组织未发现特异性染色.CDX2 mRNA在未加药物处理的Eca-109细胞株中没有表达,其表达量对DCA、GC、CA显示出时间和剂量依赖性.结论:CDX2基因异常表达在食管黏膜肠化生过程中可能具有较重要作用,CDX2基因的表达可能是Barrett’s食管发生过程的早期事件.胆汁酸在体外实验上调CDX2基因表达,通过调节其表达在Barrett’s食管发生中起重要作用.
OBJECTIVE: To observe the expression of CDX mRNA and protein in Barrett’s esophageal mucosa and to observe the effect of different bile acids on the expression of Eca-109CDX2 mRNA in human esophageal cell line.Methods: The normal esophageal tissues (n = 5), reflux Esophagitis (n = 7) and Barrett’s esophagus (n = 8). CDX1 and CDX2 mRNA expressions were determined by RT-PCR and CDX2 mRNA expression by real-time fluorescence quantitative PCR, and CDX2 protein was localized by immunohistochemistry The esophageal cancer cell line Eca-109 was cultured in 100,400,1000μmol / L cholic acid (CA), deoxycholic acid (DCA) and glycocholic acid (GC) respectively. Cells were collected at 0-24h. CDX2 mRNA expression.Results: The expression of CDX2 mRNA in normal esophageal tissues was significantly higher than that in normal esophageal tissues (9.6411 ± 6.6823,3.9156 ± 3.6700) in Barrett’s esophagus and reflux esophagitis group (P = 0.006, 0.025), but there was no significant difference between the two groups (P = 0.133), CDX1 was not expressed in all esophageal tissues.Immunohistochemical staining of CDX2 protein nuclei in Barrett’s esophagus showed specific staining Positive reaction, reflux esophagitis CDX2 cytoplasm showed fine granularity No specific staining was found in the normal esophageal tissues.CDX2 mRNA was not expressed in the untreated Eca-109 cell line, and the expression of CDX2 mRNA showed time and dose-dependent on DCA, GC and CA. Conclusion: The abnormal expression of CDX2 may play an important role in intestinal metaplasia of esophageal mucosa, and the expression of CDX2 may be an early event of Barrett ’s esophagus.The bile acid experimentally up - regulated the expression of CDX2 gene in vitro by regulating its expression in Barrett’ s Esophageal plays an important role.