Targeting microRNA-1-3p to regulate centroprotein F affects malignant development of gastric cancer through phosphoinositol 3 kinase/protein kinase B/mammalian target of rapamycin pathway
Objective To explore the effect of miR-1-3p on the malignant development of gastric cancer(GC)by targeting and regulating centroprotein F(CENPF)through phosphoinositol 3 kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)pathway.Methods Select 50 cases of gastric cancer tumor tissue and adjacent normal tissue samples treated with surgery and with surgical samples from the Department of Gastroenterology at the Second Affiliated Hospital of Jiaxing Uni-versity as the research subjects,Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of miR-1-3p in GC tissues and cells.Cell proliferation was detected by cell counting kit 8(CCK-8)and clonal formation assays.Transwell assay was used to detect cell migration and invasion.The cell cycle distribution was detected by flow cytometry.The protein expression of CENPF and PI3K/Akt/mTOR pathway was detected by Western blotting.Targeting relationship between miR-1-3p and CENPF was analyzed by dual luciferase reporter gene.Pairing design or independent sample t-test for comparison between organizations,one-way ANOVA for comparison between multiple groups.Results The miRNA expression profiles of normal and cancer tissues obtained from TCGA showed that,the expres-sion levels of miR-1-3p in gastric cancer tissues and cell lines(MGC803,AGS,MKN45,BGC-823)are lower than those in adjacent normal tissues and normal human gastric epithelial cells(5.16±1.59,0.45±0.07,0.39±0.08,0.57±0.10,0.55±0.10 vs.3.25±0.42,1.00±0.11,t=20.251,P＜0.05);The effect of miR-1-3p on the proliferation of gastric cancer cells:The cell viability of MGC803 and AGS cells transfected with miR-1-3p mimic for 72 hours was significantly lower than that of mimic NC cells(0.49±0.10,0.51±0.06 vs.0.86±0.10,0.85±0.11,t=19.205,P＜0.05);the cell viability of MGC803 and AGS cells transfected with IN-miR-1-3p for 72 hours was significantly higher than that of IN-NC cells(1.06±0.09,1.15±0.11 vs.0.83±0.10,0.85±0.11,t=23.151,P＜0.05).The effect of miR-1-3p on migration and invasion of gastric cancer cells:The number of migrating and invading cells transfected with miR-1-3p mimic in MGC803 and AGS cells was significantly lower than that in mimic NC cells(82.37±9.53,85.41±8.29,73.58±9.87,79.63±8.22 vs.138.94±15.63,140.75±16.84,114.76±10.45,121.33±9.46,t=31.012,P＜0.05);The number of migrating and invasive cells transfected with IN-miR-1-3p in MGC803 and AGS cells was significantly higher than that in IN-NC cells(192.45±16.52,203.71±17.63,158.69±11.59,162.78±14.25 vs.125.96±13.55,129.76±11.67,105.45±10.63,109.52±9.48,t=37.135,P＜0.05).The expression level of CENPF in gastric cancer tissue is higher than that in adjacent normal tissue(2.36±0.39 vs.0.98±0.10,t=28.035,P＜0.05);the expression level of CENPF in gastric cancer cell lines(AGS and MGC803)is higher than that in GES-1 cells(3.82±0.15,3.16±0.13 vs.0.99±0.08,0.70±0.05,t=17.503,P＜0.05);the CENPF levels of miR-1-3p mimic transfected into MGC803 and AGS cells were significantly lower than those in mimic NC cells(0.38±0.05,0.41±0.07 vs.1.04±0.08,1.02±0.09,t=26.611,P＜0.05);the CENPF levels in MGC803 and AGS cells transfected with IN-miR-1-3p were significantly higher than those in IN-NC cells(1.95±0.11,2.06±0.13 vs.1.03±0.07,1.00±0.08,t=28.402,P＜0.05).Overexpression of CENPF reverses the inhibitory effect of miR-1-3p mimic on CENPF:the cell viability of miR-1-3p mimic+oe NC was significantly lower than that of mimic NC+oe NC(0.93±0.12,0.88±0.12 vs.0.53±0.07,0.48±0.07,t=32.603,P＜0.05);the cell viability of miR-1-3p mimic+oe CENPF was significantly higher than that of miR-1-3p mimic+oe NC(0.90±0.09,0.91±0.11 vs.0.53±0.07,0.48±0.07,t=35.312,P＜0.05).The number of cloned cells transfected with miR-1-3p mimic+oe NC in MGC803 and AGS cells was significantly lower than that in mimic NC+oe NC(48.25±9.71,53.24±7.35 vs.103.26±12.52,116.39±14.58,t=17.157,P＜0.05);the number of cloned cells transfected with miR-1-3p mimic+oe CENPF was significantly higher than that of miR-1-3p mimic+oe NC(96.82±11.94,102.68±11.28 vs.103.26±12.52,116.39±14.58,t=19.052,P＜0.05).Conclusion MiR-1-3p negatively mediates CENPF to inhibit the malig-nant biological behavior of GC through PI3K/Akt/mTOR pathway.
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