Silencing long non-coding RNA-PVT1 inhibits the proliferation of gastric cancer cells by up-regula-ting microRNA-145
Objective The key genes long non-coding RNA(lncRNA)-PVT1 and microRNA(miR)-145 were screened in cell experiments,and the molecular mechanism of lncRNA-PVT1's role in gastric cancer was explored by silencing lncRNA-PVT1.Methods The expression level of lncRNA-PVT1 in 53 gastric cancer pathological specimens was detected by real-time fluorescence quantitative polymerase chain reaction(PCR).The expression of lncRNA-PVT1 in HGC-27 and AGS was silenced by RNA inter-ference technique.The experiments were divided into lncRNA-PVT1-SI group and NC group.The effects of lncRNA-PVT1 on the proliferation,migration and invasion of HGC-27 and AGS were detected by cell counting kit-8(CCK-8)method and Transwell cell method.The expression of miR-145 in lncRNA-PVT1-si group and NC group was detected by real-time fluorescence quantitative PCR.The direct binding sites of lncRNA-PVT1 and miR-145 were verified by immunodiluciferase assay and functional recovery assay.Real-time fluorescence quantitative PCR and Western blotting were used to detect the expression of MEST in lncRNA-PVT1-si group and NC group.x2test was used for qualitative data,and independent sample t test was used for quantitative data.Results The expression level of lncRNA-PVT1 in gastric cancer tis-sues was significantly higher than that in normal tissues(t=5.299,P＜0.05),and the expression level of lnCRNA-PVTl in HGC-27 and AGS was significantly higher than that of GES-1(t=7.714,P＜0.05;t=8.679,P＜0.05).The CCK-8 method showed that the absorbance of lncRNA-PVT1-si group was signifi-cantly lower than that of NC group at the endpoint 72 h(HGC-27 cells,t=8.054,P＜0.05;AGS cells,t=8.240,P＜0.05).Transwell migration method showed that the number of migrating cells in lncRNA-PVT1-si group was significantly decreased as compared with that in NC group(HGC-27 cells,t=5.460,P＜0.05;AGS cells,t=6.862,P＜0.05).Transwell invision method showed that the number of migrating cells in lncRNA-PVT1-si group was significantly less than that in NC group(HGC-27 cells,t=4.036,P＜0.05;AGS cells,t=7.532,P＜0.05).The expression of miR-145 in lncRNA-PVT1-si group was signifi-cantly higher than that in NC group(HGC-27 cells,t=8.381,P＜0.05;AGS cells,t=7.881,P＜0.05).Immunodiluciferase assay and functional recovery assay confirmed the direct binding site between lncRNA-PVT1 and miR-145,and the direct binding site between miR-145 and MEST.The expression of MEST in lncRNA-PVT1-si group was significantly lower than that in NC group(HGC-27 cells,t=8.821,P＜0.05;AGS cells,t=12.760,P＜0.05).Conclusion LncRNA-PVTl was up-regulated in gastric cancer cells.Knocking down lncRNA-PVT1 inhibited the proliferation,migration and invasion of gastric cancer cells by up-regulating the expression of miR-145 and then reducing the expression of MEST.
Long non-coding RNA-PVT1Gastric cancerSilenceMechanism
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