The Role And Mechanism Of LncRNA XLOC₀04787 In Gastric Cancer

Objective:Long noncoding RNA?lncRNAsNA?has emerged as a significant regulatory factor,and was reported in a variety of cancers.But the function of lncRNAsNAs in the development of malignant tumor is not completely revealed.Overall regulation of gene expression,biological role,and clinical significance of lncRNAsNAs in cancer are also not complete known.Using gene arrays,we recently found a new lncRNAsNA that may play an important role in cancer.Aiming to clarify its biological significance,we investigated the function and mechanism of action of the novel lncRNAsNA,XLOC₀04787,in gastric cancer?gastric cancer,GC?.Method:?1?Our research group used gene chip technology to screen for XLOC₀04787 with obvious expression difference.The 5'-RACE,3'-RCAE sequencing method was used to sequence the entire length of XLOC₀04787.The expression of XLOC₀04787mRNA in clinical gastric cancer samples was analyzed by real-time fluorescence quantitative PCR.Real-time quantitative PCR was used to analyze the expression of XLOC₀04787 mRNA in gastric cancer cell lines and normal gastric mucosal epithelial cells GES-1.?2?SGC-7901 was selected to transfect XLOC₀04787 small interfering RNA,and HGC-27 cells were transfected with overexpression plasmid pc DNA-XLOC₀04787;Transwell cell experiment,CCK-8 proliferation experiment,and plate clone formation experiment were used to detect the effect of XLOC₀04787 on gastric cancer cell migration and proliferation function Regulation of EMT function;Cell nuclear immunofluorescence technique was used to analyze the nuclear entry of key nuclear translocation factors p-smad2/3 and ?-catenin.?3?The lncRNAsNA prediction website was used to screen the target genes below XLOC₀04787.Dual Luciferase Reporter experiments,qRT-PCR experiments,and co-transfection experiments of cell functional effects were used to confirm targeted genes.Results:?1?qRT-PCR results showed that the expression level of XLOC₀04787 mRNA in gastric cancer tissues was relatively high?81%,26/32?compared to normal tissues.Compared with normal gastric mucosa epithelial GES-1 cells,gastric cancer cell lines are highly expressed,and the expression of SGC-7901 cells is relatively high,while the expression of HGC-27 cells is relatively low in vitro.?2?The results of Transwell chamber experiment,CCK-8 proliferation experiment,and plate clone formation experiment showed that after knocking down the expression of XLOC₀04787 in SGC-7901 cells,the cell proliferation and migration ability and EMT process were inhibited,and the Western-blot results showed that Epithelial cell marker E-Cadherin expression increased,the expression of other Mesenchymal cell markers such as N-cadherin,snail,vimentin,MMP2,and MMP9 decreased,and the expression of key indicators in the Wnt and TGF-?signaling pathways decreased,and the results of cell immunofluorescence experiments showed that the nuclear translocation tendency of key factors p-smad2/3and ?-Catenin decreased.After transfection of the overexpression plasmid pc DNA-XLOC₀04787 to increase the expression of XLOC₀04787 in HGC-27 cells,compared with the control groups,the cell's migration and proliferation ability was enhanced and the cell's EMT process was induced.The Western-blot results showed that the expression of Epithelial cell marker E-cadherin decreased and other Mesenchymal cell markers such as N-cadherin,snail,vimentin,MMP2,and MMP9 were all increased,and the expression of key indicators in the Wnt and TGF-?signaling pathways increased,and the results of immunofluorescence experiments found that the nuclear translocation tendency of key factors p-smad2/3 and ?-Catenin increased.?3?The prediction website predicts the downstream target gene miR-203a-3p.Through qRT-PCR results,it was found that the expression of miR-203a-3p increased after knocking down the expression of miR-203a-3p,and the expression of miR-203a-3p increased after overexpression of XLOC₀04787.The double luciferase reporter gene experiment found that the ratio of firefly / renilla luciferase in the experimental group transfected with the XLOC₀04787 wild-type plasmid and miR-203a-3p-mimics decreased compared to other experimental groups.And miR-203a-3p-mimics and XLOC₀04787-siRNA co-transfection functional experiments found that cell proliferation and migration inhibition were alleviated in the co-transfection group compared to the XLOC₀04787 knockdown group.Conclusion:?1?The expression level of XLOC₀04787 is higher in gastric cancer tissues and gastric cancer cell lines.?2?Highly expressed XLOC₀04787 can promote the proliferation and migration of gastric cancer cells and the EMT process.?3?XLOC₀04787 plays a role in promoting cancer in gastric cancer cells,and may provide new targeting molecules for the diagnosis and treatment of gastric cancer.