Kruppel样转录因子5对食管鳞状上皮转分化绒毛蛋白表达的影响  

Kruppel-like factor 5 regulates villin expression in intestinal transdifferentiation of esophageal squamous epithelium

作　　者：夏一菊[1] 张昊翔[1] 沈才飞[1] 王璞[1] 闫武[1] 李婧文[1] 徐胤[1] 房殿春[1] 

机构地区：[1]第三军医大学附属西南医院消化科,重庆400038

出　　处：《中华消化杂志》2016年第4期265-270,共6页Chinese Journal of Digestion

基　　金：国家自然科学基金（81170356、81270450）

摘　　要：目的 探讨脱氧胆酸诱导食管鳞状上皮细胞肠化生过程中，Krüppel样转录因子（KLF）5对绒毛蛋白的调节作用。 方法 采用免疫组织化学法检测KLF5和绒毛蛋白在人正常食管鳞状上皮组织（20份）、反流性食管炎组织（59份）、Barrett食管组织（21份）中的表达；使用HET-1A细胞，分别用脱氧胆酸处理2、4、8、12 h，KLF5 RNA干扰（小干扰RNA）下调和慢病毒过表达KLF5后，PCR、蛋白质印迹法检测其KLF5、绒毛蛋白mRNA及其蛋白质的表达。计数资料采用卡方检验比较，计量资料采用配对t检验分析比较。 结果 免疫组织化学法显示，KLF5、绒毛蛋白在正常食管黏膜未见表达，在食管炎组织呈部分弱表达[分别为27.1%（16/59）和20.3%（12/59）]，而在Barrett食管组织中表达明显增强[分别为85.7%（18/21）和100.0%（21/21），差异均有统计学意义（χ2＝36.576、56.765, P均〈0.01）]。200 μmol/L脱氧胆酸分别处理HET-1A细胞2 h后，与未加脱氧胆酸的对照组（0 h）相比，KLF5、绒毛蛋白mRNA（t＝－16.64, －11.85）及其蛋白质（t＝－30.80, －20.70）表达量增加（P均〈0.01）。采用RNA干扰技术下调KLF5表达可阻止胆酸诱导的绒毛蛋白表达，采用基因转染技术上调KLF5表达后，可同时上调绒毛蛋白在HET-1A细胞中的表达。 结论 在胆酸介导的食管鳞状上皮细胞肠化生过程中，KLF5对绒毛蛋白表达具有调节作用，可能参与Barrett食管的发生。Objective To investigate the regulatory role of Kruppel-like factor 5 （KLF5） in villin expression during the process of deoxycholic acid （DCA） induced intestinal transdifferentiation of esophageal squamous epithelium. Methods The expressions of KLF5 and villin in normal esophageal epithelial tissues （20 cases）, reflux esophagitis tissues （ 59 cases） and Barrettt s esophagus tissues （21 cases） were detected by immunohistochemistry. After the HET-1A cells treated by DCA for two, four, eight, 12 h, KLF5 were down regulated by small interfering （si）RNA and over-expressed by lentivirus, and then the expressions of KLF5 and villin at mRNA and protein level were measured by polymerase chain reaction （PCR） and Western blotting. The counting data were by Chi-square test, and the measurement data were compared by t test. Results The results of immunohistochemistry showed that there was no expressions of villin and KLF5 in normal esophageal squamous epithelium, partial weak expressions in esophagitis tissue （27.1%（16/59） and 20.3%（12/59）, respectively） and strong expressions in Barrett＇s esophagus tissue （85. 7% （18/21） and 100. 0% （21/21））, respectively, and the differences were statistically significant （x2=36. 576, 56. 765, both P〈0.01）. After HETdA cells treated by 200 μmol/L DCA for 12 h, compared with the control （treated with non DCA）, the expression of KLF5 and villin at mRNA （t=-16.64, -11.85） and protein levels （t=-30.80, --20. 70） increased （all P〈0.01）. Once theexpression of KLF5 downregulated by siRNA, DCA induced expression of villin was inhibited. The expression of KLF5 was up regulated by transfection, which also increased the expression of villin in HET-1A cells. Conclusion In DCA mediated intestinal transdifferentiation of esophageal squamous epithelium, KLF5 plays a role in the regulation of villin expression and may be involved in the genesis of Barrett＇s esophagus.

关 键 词：BARRETT食管 Kriippel样转录因子5 绒毛蛋白 化生 

分 类 号：R735.1[医药卫生—肿瘤] R571[医药卫生—临床医学]