MiR-590-5p Regulates The Sensitivity Of Human Gastric Cancer Cells To Cisplatin By Targeting PDCD4

Objective :MiR-590-5p is dysregulated in a variety of tumors,thereby affecting tumors' biological behaviors such as proliferation,invasion,and migration.Reading the literature and previous experimental results show that 590-5p is associated with cisplatin resistance in gastric cancer,but the specific mechanism has not been clarified.This article mainly explores miR-590-5p regulates the sensitivity of human gastric adenocarcinoma cells to cisplatin through downstream target genes and its specific molecular mechanism.Methods: The relative expression of miR-590-5p relative to normal gastric mucosal cells in gastric adenocarcinoma cells SGC7901,BGC823 and AGS was detected by real-time fluorescent quantitative PCR.Bioinformatics software such as Targetscan was used to predict the downstream target gene of miR-590-5p,and the luciferase reporting experiment was used to verify the binding of miR-590-5p to the downstream target gene.The liposomes Lipofectamine 2000 were used to transfect miR-590-5p mimics and miR-590-5p inhibitor to human gastric adenocarcinoma cells with low and high expression of miR-590-5p,respectively,and verified by Western blot and qRT-PCR.Expression of downstream target genes;flow cytometry was used to detect changes in apoptosis of cells under the action of cisplatin;MTT detected gastric cancer cells transfected with different cisplatin concentrations(0,0.5,1,2,4,8)?mol/L sensitivity changes.In addition,in order to verify the effect of the target gene,the target gene was knocked out and Western blot was used to verify the expression of the target gene;flow cytometry was used to detect changes in the apoptosis of cells under the action of cisplatin;MTT was used to detect gastric cancer cells after transfection Changes in sensitivity to different cisplatin concentrations(0,0.5,1,2,4,8)?mol/L.Results: The results of real-time quantitative PCR showed that miR-590-5p expression was lower in SGC7901 cells than in BGC823 cells.Bioinformatics software predicts that miR-590-5p and PDCD4 can be targeted for complementarity,and luciferase experiments confirm that miR-590-5p and wild-type PDCD43'UTR bind luciferase activity is significantly reduced.Western blot and qRT-PCR indicated that SGC7901 cells and BGC823 over-expressed and inhibited miR-590-5p,respectively,and PDCD4 expression decreased and increased accordingly(p <0.05);flow cytometry showed that SGC7901 cells and BGC823 over-expressed and After miR-590-5p was inhibited,apoptosis decreased and increased correspondingly(p<0.05);human gastric adenocarcinoma cell SGC7901 up-regulated miR-590-5pmimics had higher sensitivity to cisplatin than parental cell SGC7901,and transfected miR BGC823 cells after-590-5p inhibitor had reduced sensitivity to cisplatin(p <0.05).After knocking out the target gene PDCD4,its protein expression decreased(p <0.05),flow cytometry results showed that gastric cancer cells had reduced apoptosis(p <0.05),and MTT results showed reduced sensitivity to cisplatin(p <0.05).Conclusion : miR-590-5p may affect the sensitivity of gastric cancer cells to cisplatin by regulating the expression of the downstream target gene PDCD4.miR-590-5p down-regulates the expression of target gene PDCD4;PDCD4 may affect the sensitivity of gastric cancer cells to cisplatin by promoting the apoptosis of gastric cancer cells.