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目的:探究微小RNA(miR)-1-3p靶向调控中心粒蛋白F(CENPF)通过磷酸肌醇3激酶(PI3K)/蛋白激酶B(Akt)/哺乳动物雷帕霉素靶蛋白(mTOR)通路影响胃癌(GC)恶性发展的机制。方法:选取嘉兴大学附属第二医院胃肠外科收治的50例经手术治疗、且留有手术样本的胃癌肿瘤组织及邻近正常组织样本作为研究对象，采用实时实时荧光定量聚合酶链反应(qRT-PCR)检测胃癌组织及细胞中miR-1-3p的表达。细胞计数试剂盒-8(CCK-8)及克隆形成实验检测细胞增殖；Transwell检测细胞迁移及侵袭能力；流式细胞术检测细胞周期分布；蛋白质印迹法(Western blot)检测CENPF及PI3K/Akt/mTOR通路蛋白表达；双荧光素酶报告基因分析miR-1-3p和CENPF的靶向关系。组织之间的比较行配对设计或独立样本 t检验，多组间的比较行单因素方差分析。 结果:从TCGA获取正常组织和癌症组织的miRNA表达谱，结果发现，胃癌组织及细胞系中(MGC803、AGS、MKN45、BGC-823)的miR-1-3p表达水平底于邻近正常组织及正常人胃上皮细胞(5.16±1.59、0.45±0.07、0.39±0.08、0.57±0.10、0.55±0.10比3.25±0.42、1.00±0.11， t＝20.251， P<0.05)；miR-1-3p对胃癌细胞增殖的影响：MGC803和AGS细胞转染miR-1-3p mimic 72 h的细胞活力明显低于mimic NC(0.49±0.10、0.51±0.06比0.86±0.10、0.85±0.11， t＝19.205， P<0.05)；MGC803及AGS细胞转染IN-miR-1-3p 72 h的细胞活力明显高于IN-NC(1.06±0.09、1.15±0.11比0.83±0.10、0.85±0.11， t＝23.151， P<0.05)。miR-1-3p对胃癌细胞迁移侵袭的影响：MGC803和AGS细胞转染miR-1-3p mimic的迁移细胞数及侵袭细胞数明显低于mimic NC(82.37±9.53、85.41±8.29、73.58±9.87、79.63±8.22比138.94±15.63、140.75±16.84、114.76±10.45、121.33±9.46， t＝31.012， P<0.05)；MGC803及AGS细胞转染IN-miR-1-3p的迁移细胞数和侵袭细胞数明显高于IN-NC(192.45±16.52、203.71±17.63、158.69±11.59、162.78±14.25比125.96±13.55、129.76±11.67、105.45±10.63、109.52±9.48， t＝37.135， P<0.05)。CENPF在胃癌组织中的表达量高于邻近正常组织(2.36±0.39比0.98±0.10， t＝28.035， P<0.05)；CENPF在胃癌细胞系中(AGS和MGC803)的表达量高于GES-1细胞(3.82±0.15、3.16±0.13比0.99±0.08、0.70±0.05， t＝17.503， P<0.05)；MGC803及AGS细胞转染miR-1-3p mimic的CENPF水平明显低于mimic NC(0.38±0.05、0.41±0.07比1.04±0.08、1.02±0.09， t＝26.611， P<0.05)；MGC803及AGS细胞转染IN-miR-1-3p的CENPF水平明显高于IN-NC(1.95±0.11、2.06±0.13比1.03±0.07、1.00±0.08， t＝28.402， P<0.05)。CENPF过表达逆转miR-1-3p mimic对CENPF的抑制作用，MGC803及AGS细胞转染72 h后，miR-1-3p mimic＋oe-NC的细胞活力明显低于mimic-NC＋oe-NC(0.93±0.12、0.88±0.12比0.53±0.07、0.48±0.07， t＝32.603， P<0.05)；miR-1-3p mimic＋oe-CENPF的细胞活力明显高于miR-1-3p mimic＋oe-NC(0.90±0.09、0.91±0.11比0.53±0.07、0.48±0.07， t＝35.312， P<0.05)。克隆形成实验结果显示，MGC803及AGS细胞转染miR-1-3p mimic＋oe-NC的克隆细胞数量明显低于mimic-NC＋oe-NC(48.25±9.71、53.24±7.35比103.26±12.52、116.39±14.58， t＝17.157， P<0.05)；转染miR-1-3p mimic＋oe-CENPF后克隆细胞数量明显高于miR-1-3p mimic＋oe-NC(96.82±11.94、102.68±11.28比103.26±12.52、116.39±14.58， t＝19.052， P<0.05)。 结论:miR-1-3p负向调控CENPF通过PI3K/Akt/mTOR通路阻碍了GC的恶性生物学行为。

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 University 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 expression 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 malignant biological behavior of GC through PI3K/Akt/mTOR pathway.