Expression Of LINC00261 In Barrett's Esophagus And Esophageal Adenocarcinoma And Its Possible Mechanism

To evaluate the expression of LINC00261 in Barrett's esophagus and esophageal adenocarcinoma and to investigate the effect of LINC00261 expression on the differentiation phenotype of embryonic stem cells induced by acid and bile exposure and its possible mechanism.The following methods are discussed: RecoverAll? Total Nucleic Acid Isolation Kit was used to extract total RNA from FFPE Tissues(20 cases of normal esophageal squamous tissue,20 cases of esophageal adenocarcinoma tissue and 20 cases of Barrett esophagus tissue),and qRT-PCR was used to analyze the expression changes of LINC00261 in normal esophageal squamous epithelium,Barrett's esophagus and esophageal adenocarcinoma tissues.Make the four different cells line to inoculate respectively in R PMI-1640 medium until cells into the logarithmic phase.Extract total RNA from cells,and qRT-PCR was used to analyze the expression changes of LINC00261.Human embryonic stem cells were cultured on the feeder layer of human embryonic fibroblasts until into the logarithmic grow phase and were exposed to different concentrations of acid and bile.The total RNA was extracted after treated with 24 h,48h and 72 h,and qRT-PCR was used to analyze the expression changes of LINC00261.Analysis of downstream genes regulated by LINC00261 by Bioinformatics.qRT-PCR was used to analyze the expression changes of lentivirus-mediated LINC00261 and Western blot was used to analyze the expression changes of Lentivirus-mediated FOXA2.Western blot was used to analyze the expression changes of CDX2 and MUC2 in different expression of FOXA2 in embryonic stem cells.SPSS 17 statistical software was used to analyze the results of the experiment.P<0.05 showed the difference was significant.We found the expression of LINC00261 in Barrett's esophagus and esophageal adenocarcinoma were both significantly higher than in normal esophageal squamous epithelium tissues and cells(P<0.01).Acid and bile exposure could increase the expression of LINC00261 and showed a concentration-dependent and time-dependent manner.Bioinformatics revealed LINC00261 locate in the upstream of FOXA2 and participate the expression of FOXA2.Lentivirus-mediated LINC00261 upregulation could induce the inhibition of FOXA2 protein expression.Acid and bile exposure could induce the inhibition of FOXA2 protein expression,what's more,induce the expression of CDX2(a biomarker for intestinal differentiation)and MUC2(a biomarker for columnar differentiation).Lentivirus-mediated FOXA2 upregulation could attenuate the influence of acid and bile exposure in human embryonic stem cells on the expression of CDX2(a biomarker for intestinal differentiation)and MUC2(a biomarker for columnar differentiation).In general,LINC00261 plays a key role in the development of Barrett's esophagus,which might be related to its negative regulation effect on FOXA2 gene transcription and induce the expression of CDX2 and MUC2.