POU3F3对垂体瘤细胞增殖和周期的影响及其作用机制  

Effect of POU3F3 on proliferation and cell cycle of pituitary adenoma cells and its mechanism

作　　者：王纵 李力 王莹 Wang Zong;Li Li;Wang Ying(Department of Neurosurgery,Zhumadian Central Hospital Affiliated to Huanghuai University,Zhumadian 463000,China;Department of Neurosurgery,Affiliated Tumor Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]黄淮学院附属驻马店市中心医院神经外科,463000 [2]郑州大学附属肿瘤医院神经外科,450000

出　　处：《中华实验外科杂志》2020年第9期1614-1617,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨POU3F3对垂体瘤细胞增殖和周期的影响及其作用机制。方法选取2016年6月到2019年12月驻马店市中心医院手术切除的150例垂体瘤和癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)分析POU3F3和微小RNA(miRNA,miR)-127-5p表达水平;采用短发卡RNA(shRNA)和过表达技术在RC-4BC垂体瘤细胞中分别敲降POU3F3和过表达miR-127-5p,采用Lipo2000过表达采用细胞计数试剂盒(CCK-8)和流式细胞术分析POU3F3和miR-127-5p对细胞增殖和周期的影响;采用生物信息学和双荧光素酶报告基因分析POU3F3和miR-127-5p及其靶基因关系;采用蛋白质印迹法(Western blot)分析POU3F3和miR-127-5p对靶基因表达的影响。组间比较采用t检验。结果与癌旁组织中POU3F3表达水平(1.10±0.19)比较,垂体腺癌组织中POU3F3表达水平(2.84±0.22)显著增加,差异有统计学意义。与癌旁组织中miR-127-5p表达水平(1.04±0.15)比较,垂体腺癌组织中miR-127-5p表达水平(0.34±0.11)显著下调,差异有统计学意义。与对照shRNA组细胞G0/G1期、S期和G2/M期[(37.54±3.45)%、(39.10±3.90)%和(20.58±2.87)%]比较,POU3F3 shRNA组细胞G0/G1期比例(64.33±6.24)%显著增加,而S期和G2/M期比例[(20.14±3.01)%和(14.33±2.88)%]显著下调,差异有统计学意义。与对照mimic组细胞G0/G1期、S期和G2/M期[(35.66±4.02)%、(40.89±4.61)%和(25.33±2.19)%]比较,miR-127-5p minic组细胞G0/G1期比例显著增加[(59.48±6.14)%],而S期和G2/M期比例[(24.23±2.71)%和(17.59±1.99)%]显著下调,差异有统计学意义。miR-127-5p与POU3F3存在碱基配对,FOXD1是miR-127-5p靶基因。与对照shRNA组和对照mimic组细胞FOXD1蛋白表达水平(1.16±0.21、0.97±0.14)比较,POU3F3 shRNA组和miR-127-5p mimic组细胞FOXD1蛋白表达水平(0.22±0.15、0.32±0.12)显著下降,差异有统计学意义。结论POU3F3通过miR-127-5p/FOXD1调控垂体腺癌细胞增殖和周期。Objective To investigate the effect of POU3F3 on the proliferation and cell cycle of pituitary tumor cells and its mechanism.Methods 150 cases of pituitary adenomas and adjacent objects were selected as research objection.The expression levels of POU3F3 and microRNA(miRNA,miR)-127-5p were analyzed by fluorescence quantitative polymerase chain reaction(PCR).POU3F3 were knocked down in RC-4BC pituitary tumor cells by short hairpin RNA(shRNA)technology and miR-127-5p were overexpressed in RC-4BC pituitary tumor cells.The proliferation and cell cycle were analyzed by cell counting kit-8(CCK-8)and flow cytometry.The relationship between POU3F3 and miR-127-5p were analyzed bioinformatics and double luciferase reporter genes.The expression of target genes in POU3F3 and miR-127-5p were analyzed by Western blotting.Results Compared with the expression level of POU3F3(1.10±0.19)in adjacent tissues,the expression of POU3F3 in pituitary adenocarcinoma significantly increased(2.84±0.22).Compared with the expression level of miR-127-5p(1.04±0.15)in adjacent tissues,the expression level of miR-127-5p in pituitary adenocarcinoma tissue significantly decreased(0.34±0.11).Compared with the control shRNA group,the proportion of G0/G1 phase,S phase and G2/M phase[(37.54±3.45)%,(39.10±3.90)%and(20.58±2.87)%],the proportion of G0/G1 phase in POU3F3 shRNA group significantly increased(64.33±6.24)%,while the ratio of S phase and G2/M phase[(20.14±3.01)%and(14.33±2.88)%]significantly decreased in POU3F3 shRNA group.Compared with the control group,the proportion of G0/G1 phase,S phase and G2/M phase in miR-127-5p Mimic group significantly increased[(59.48±6.14)%],while the proportion of S phase and G2/M phase[(24.23±2.71)%and(17.59±1.99)%]in miR-127-5p mimic group significantly reduced.POU3F3 could complement and bind to miR-127-5p and miR-127-5p regulated the expression of FOXD1 protein.The expression level of FOXD1 protein in POU3F3 shRNA group and miR-127-5p mimic Group(0.22±0.15,0.32±0.12)were significantly lower than t

关 键 词：POU3F3 微小RNA-127-5p FOXD1 垂体腺癌 增殖 周期 

分 类 号：R736.4[医药卫生—肿瘤]