LncRNA CTA-250d10.23 In The Role Of Esophageal Squamous Carcinoma And Molecular Mechanism Research

Objective: In esophageal squamous cell carcinoma and adjacent tissues of testing to determine the long chain noncoding RNA CTA-250d10.23 expression differences,and explore its role in esophageal squamous cell carcinomas,provide new therapeutic targets for clinical treatment.Methods: 1.Collect esophageal squamous cell carcinoma and adjacent tissues,and through the qRT-PCR detection of lncRNA CTA-250d10.23 expression differences;And statistical esophageal squamous cell carcinoma patients with clinical pathological information;The qRT-PCR detection of LncRNA expression in normal esophageal mucosa epithelial cells(Het-1 a)and esophageal squamous cancer cells(TE-1,TE-10,TE-11,YSE140,KYSE150,EC109 and EC9706).According to the expression of LncRNA,follow-up experiments needed for cell screening.2.The high expression of lncRNA CTA-250d10.23 KYSE150,TE-11 cell line,through the siRNA interference technology to decrease the lncRNA expression,then use the CCK 8 method is used to detect the proliferation of esophageal squamous cell carcinoma cell;the effects of using scratches experiment,transwell experimental detection the migration ability in esophageal squamous cell carcinomas cell,which lncRNA expression was interfered by siRNA interference technology;And by building lncRNA CTA-250d10.23 expression plasmid,further test itsfunctions in cell proliferation and migration of esophageal squamous cell carcinoma cell.3.Through the literature and bioinformatics prediction found there was a link between miR-326 and lncRNA CTA-250d10.23,the siRNA interference lncRNA CTA-250d10.23 expression,can release the micro miR-326 expression;And explore has-miR-326 impact of esophageal squamous cell carcinoma of the biological behaviors.4.To explore the lncRNA CTA-250d10.23 in the role of esophageal squamous cell carcinoma,using bioinformatics forecast micro miR-326 downstream target genes,further qRT-PCR and western blotting technology test of downstream target genes mRNA and protein expression level of the situation.5.Use Graph pad Prism 6.0statistical analysis of experimental results data and drawing,p value is less than0.05,it has statistical significance.Results:1.The expression of long noncoding RNA CTA-250d10.23 in esophageal squamous cell carcinoma and paracancerous tissues was significantly higher than that in paracancerous tissues.The clinicopathological information of patients with esophageal squamous cell carcinoma was statistically analyzed,found the differently expressed of CTA-250d10.23 was not significantly related to the age,gender,depth of tumor invasion,size,grade and stage of esophageal squamous cell carcinoma patients;meanwhile,the expression of lncRNA CTA-250d10.23 was increased in esophageal squamous cell carcinoma cells,among which KYSE150 and TE-11 cells showed the greatest difference as follow-up experimental cells.2.The results of CCK-8showed that after interfering with the expression of long noncoding RNACTA-250d10.23,it could inhibit the proliferation of cells;Scratch test and Transwell cell lab test showed that after interfering with long noncoding expression,the migration of cells was inhibited;After transfection with over expression plasmids,the proliferation and migration of KYSE150 and TE-11 cells were improved.3.Long noncoding RNA CTA-250d10.23 can affect the expression of has-miR-326;transfection of has-miR-326 inhibitor can increase the proliferation and migration of KYSE150 cell line,while transfection of has-miR-326 mimic can inhibit the proliferation and migration of cells.4.The target gene of has-miR-326 was found by bioinformatics prediction and literature review.After the target gene was transfected with the inhibitor,the inhibition of microRNA on the target gene mRNA was relieved.At the same time,the mRNA level of the target gene was inhibited by the transfection of has-miR-326 mimic.In KYSE150 cells,there was lncRNA CTA-250d10.23 can inhibit the expression of downstream target genes,especially the expression of RARG.At the same time,at the protein level,the expression of target gene protein decreased after transfection of has-mir-326 mic,and the expression of RARG protein increased after the addition of inhibitor,and the overexpression of lncRNA CTA-250d10.23.Conclusion:lncRNA CTA-250d10.23 can restrain through targeted micro has-miR-326 activation RARG expression,thus promotes the proliferation of esophageal squamous cell carcinoma,migration and other biological behaviour.