肝细胞核因子4α在胆汁酸诱导的胃黏膜肠上皮化生中的作用及其机制  

Role and mechanisms of hepatocyte nuclear factor 4α in bile acid induced gastric intestinal metaplasia

作　　者：闵亚莉 倪阵[2] 雷超[2] 袁挺 杜风[2] 刘凯歌[3] 时永全[2] Min Yali;Ni Zhen;Lei Chao;Yuan Ting;Du Feng;Liu Kaige;Shi Yongquan(Clinical Medical College, Ningxia Medical University, Yinchuan 750004, Chin)

机构地区：[1]宁夏医科大学临床医学院,银川750004 [2]第四军医大学西京消化病医院消化科 [3]西安医学院第一附属医院消化科

出　　处：《中华消化杂志》2018年第3期165-170,共6页Chinese Journal of Digestion

基　　金：国家自然科学基金（81470805）

摘　　要：目的探讨肝细胞核因子4α（HNF4α）在鹅脱氧胆酸（CDCA）诱导的胃黏膜肠上皮化生中的作用与机制。 方法以不同浓度的CDCA刺激人永生化胃黏膜上皮细胞GES-1后，采用实时荧光定量PCR和蛋白质印迹法检测GES-1和人胃癌细胞系（AGS、SGC7901、BGC823）中HNF4α、尾侧同源盒转录因子2（CDX2）和三叶肽因子家族3（TFF3）的mRNA和蛋白质表达变化。用HNF4α短发夹RNA和对照短发夹RNA分别转染GES-1后，以CDCA刺激，采用蛋白质印迹法检测HNF4α、CDX2和TFF3的蛋白质表达。采用慢病毒感染的方法，分别在GES-1和SGC7901中过表达HNF4α，在BGC823和AGS中静默HNF4α，以实时荧光定量PCR和蛋白质印迹法检测HNF4α、CDX2、TFF3的mRNA和蛋白质表达。采用荧光素酶报告基因实验分析HNF4α对CDX2的调控作用。统计学分析采用t检验。 结果GES-1、SGC7901、BGC823和AGS的HNF4α mRNA水平分别为1.00±0.12、263.01±10.23、848.01±18.13和3 049.86±91.75；其中AGS、BGC823和SGC7901的HNF4α mRNA水平均高于GES-1，差异均有统计学意义（t＝33.23、46.72、25.62，P均〈0.01）；AGS、BGC823、SGC7901和GES-1的HNF4α蛋白质表达与mRNA表达一致。转染HNF4α短发夹RNA组的CDX2和TFF3蛋白质表达低于未转染HNF4α短发夹RNA组。在GES-1细胞中，过表达HNF4α组HNF4α、CDX2和TFF3的mRNA水平分别为16 281.839±1 843.017、6.275±0.137和17.310±1.533，分别高于过表达对照组的1.000±0.048、1.000±0.012和1.000±0.108，差异均有统计学意义（t＝8.83、38.29、10.61，P均〈0.01）；在AGS细胞中，静默HNF4α组HNF4α、CDX2和TFF3的mRNA水平分别为0.021±0.001、0.088±0.007和0.074±0.002，分别低于静默对照组的1.000±0.108、1.000±0.131和1.000±0.122，差异均有统计学意义（t＝9.09、6.93、7.57，P均〈0.01）；在GES-1过表达细胞和AGS静默细胞中，HNF4α、CDX2、TFF3的蛋白质表达与mRNA表达一致。在克隆有CDX2-1（－2 000～－1 bp）�ObjectiveTo investigate the roles and mechanisms of hepatocyte nuclear factor 4α （HNF4α） in chenodeoxycholic acid （CDCA） induced gastric intestinal metaplasia （IM）. MethodsAfter the immortalized gastric mucosal epithelial cells GES-1 were stimulated with CDCA at different concentration, the changes of HNF4α, caudal-related homeobox 2 （CDX2） and trefoil factor family 3 （TFF3） expressions at mRNA and protein levels in GES-1 cells and gastric cancer cell lines （AGS, SGC7901 and BGC823） were detected by real time-polymerase chain reaction （RT-PCR） and Western blotting. After GES-1 were transfected with HNF4α short hairpin RNA （shRNA） or control shRNA, and followed by CDCA stimulation, the expressions of HNF4α, CDX2 and TFF3 at protein level were determined by Western blotting. HNF4α was overexpressed in GES-1 cells and SGC7901 cells, and HNF4α was silenced in BGC823 cell line and AGS by lentiviral vector system. The expressions of HNF4α, CDX2 and TFF3 at mRNA and protein levels were tested by RT-PCR and Western blotting. Luciferase reporter assay was used to analyze the regulation role of HNF4α on CDX2. T test was performed for statistical analysis. ResultsThe expressions of HNF4α in GES-1, SGC7901, BGC823 and AGS cells at mRNA level were 1.00±0.12, 263.01±10.23, 848.01±18.13 and 3 049.86±91.75, respectively. The mRNA levels of HNF4α in AGS, BGC823 and SGC7901 cells were all higher than that of GES-1 cells, and the differences were statistically significant （t=33.23, 46.72 and 25.62, all P〈0.01）. The expressions of HNF4α in GES-1, SGC7901, BGC823 and AGS at protein level were consistent with mRNA level. The expressions of CDX2 and TFF3 at protein level of HNF4α shRNA transfected group were lower than those of non-HNF4α shRNA transfected group. In GES-1 cells, the expressions of HNF4α, CDX2 and TFF3 of HNF4α overexpressed group at mRNA level were 16 281.839±1 843.017, 6.275±0.137 and 17.310±1.533, respectively; which were all higher than those of overe

关 键 词：肝细胞核因子4 肝细胞核因子4Α 肠上皮化生 尾型同源盒转录因子-2 

分 类 号：R735.2[医药卫生—肿瘤]