MiR-105-5p regulates fatty acid metabolism and biological behaviors of gastric cancer cells by targeting SIRT1
Objective To investigate the effects of miR-105-5p on the proliferation,invasion,migration,and fatty acid metabolism of gastric cancer cells,and to elucidate the targeted relationship between miR-105-5p and silent information regulatory factor 1(SIRT1)gene.Methods Human gastric mucosa epithelial cells GES-1 and human gastric cancer HepG2 cells were selected.Different plasmids were transfected into HepG2 cells and they were divided into miR-105-5p group(transfected with miR-105-5p mimic),si-miR-105-5p group(transfected with miR-105-5p inhibitor),miR-NC group(transfected with mimic NC),NC group(transfected with inhibitor NC),Scramble group(transfected with Scramble),miR-105-5p+Scramble group(transfected with miR-105-5p mimic and Scramble)and miR-105-5p+sh-SIRT1 group(transfected with miR-105-5p mimic and sh-SIRT1).MTT assay was used to detect cell proliferation.Transwell assay was used to detect cell migration and invasion.The enzyme-linked immunosorbent assay(ELISA)was used to detect the content of phospholipid and triacylglycerol.The real-time quantitative PCR(RT-qPCR)was used to detect the mRNA levels of miR-105-5p and SIRT1 in cells.Western blot was used to detect the protein levels of SIRT1,fatty acid synthase(FASN),Acetyl-CoA carboxylase 1(ACC1),fatty acid binding protein 5(FABP5)in cells.The targeting relationship between miR-105-5p and SIRT1 was predicted by bioinformatics,and verified by dual-luciferase reporter assay.Results The level of miR-105-5p in HepG2 cells was higher than that in GES-1 cells,while the mRNA and protein levels of SIRT1 were significantly lower than those in GES-1 cells(P<0.01).The optical density(OD)values at 48h and 72h,invasive and migratory cell numbers,phospholipid content,triacylglycerol content,and protein levels of FASN,ACC1 and FABP5 in the miR-105-5p group were significantly higher than those of the miR-NC group,which,in the si-miR-105-5p group were significantly lower than those of the NC group(P<0.01).OD values at 48h and 72h,invasive and migratory cell numbers,phospholipid content,triacylglycerol content,and protein levels of FASN,ACC1 and FABP5 in the miR-105-5p + Scramble group were significantly higher than those of the Scramble group,which,in the miR-105-5p+sh-SIRT1 group were significantly lower than those of the miR-105-5p+ Scramble group(P<0.01).The protein level of SIRT1 in the miR-105-5p group was significantly lower than that of the miR-NC group,which,in the si-miR-105-5p group was significantly higher than that of the NC group(P<0.01).MiR-105-5p had a complementary base pairing site with SIRT1 3'UTR.The luciferase activity in miR-105-5p+WT-SIRT1 group was significantly lower than that of miR-NC+WT-SIRT1 group(P<0.01).Conclusion MiR-105-5p may inhibit the proliferation,invasion,and migration of gastric cancer cells by regulating fatty acid metabolism through SIRT1.
miR-105-5psilent information regulatory factor 1gastric cancerfatty acid metabolisminvasion
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