微小RNA-23通过靶向调节SRY相关高迁移率族盒蛋白-5的表达显著抑制脑垂体腺瘤细胞的增殖  

MicroRNA- 23 significantly inhibits the proliferation of pituitary adenoma cells by targeting the gene SRY - related high - mobility - group box - 5

作　　者：李永明[1] 赵亚军 秦涛[2] Li Yongming;Zhao Yajun;Qin Tao(Department of Neurosurgery,Henan Provincial People＇s Hospital,Zhengzhou 450003,China（Li YM,Zhao YJ;Department of Hepatobiliary Surgery,Henan Provincial People＇s Hospital,Zhengzhou 450003,China（Qin)

机构地区：[1]河南省人民医院神经外科,郑州450003 [2]河南省人民医院肝胆外科,郑州450003

出　　处：《中华实验外科杂志》2018年第10期1898-1901,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察微小RNA（miRNA，miR）乞3靶向调控SRY相关高迁移率族盒蛋白-5（SOX5）的表达及其对脑垂体腺瘤细胞增殖的影响。方法将miR-23mimic及阴性对照（NC）采用Lipofectamine脂质体法转染至脑垂体腺瘤细胞，并记为miR-23mimic组和NC组，以未转染的脑垂体腺瘤细胞作为对照组。转染后，采用实时荧光定量聚合酶链反应（VQ．PCR）法检测各组细胞miR-23和SOX5mRNA的表达量，Westernblot检测各组细胞SOX5的蛋白水平，噻唑蓝（MTI）法检测的各组细胞增殖活性，采用荧光素酶报告实验检测miR-23对SOX5的靶向调控作用，过表达SOX5与miR-23mimic共转染后检测细胞增殖活性。结果FQ-PCR检测结果显示，转染48h后miR-23mimic组的miR-23的表达水平（0．54±0．13）低于对照组（1．00，P=0．001）和NC组（1．03±0．09，P=0．000），差异有统计学意义；miR-23mimic组增殖活性与其余两组比较，转染24、48、72、96h后的脑垂体腺瘤细胞的增殖能力均降低（A24h=0．47±0．04，A48h=0．52±0.05，A72 h=0．54±0．05，A96h=0．55±0．06，与对照组比较，P1=0．036，P2=0．019，P=0．006，P4=0．000）。荧光素酶报告基因实验表明miR-23可靶定结合SOX5mRNA3’端非编码区（3’UTR）。PcDNA-SOX5与miR-23mimic共转染后，经细胞增殖检测显示，过表达SOX5可抵抗miR-23mimic引起的细胞增殖抑制作用。结论miR-23可靶向下调SOX5的表达显著抑制脑垂体腺瘤细胞的增殖。Objective To investigate the expression of SRY - related high - mobility - group box - 5 （ SOX5 ） by microRNA - 23 （ miR - 23 ） and its effect on the proliferation of pituitary adenoma cells. Methods MiR - 23 mimic and negative control （NC） were transfeeted into pituitary adenoma cells by Lipofectamine liposome method and recorded as miR -23 mimic group and NC group. Untransfeeted pituitary adenoma cells were used as Control group. After transfection, the expression levels of miR -23 and SOX5 in each group were detected by real - time fluorescent quantitative polymerase chain reaction （ FQ - PCR）. The protein levels of SOX5 in each group were detected by Western blotting, and the prolif- eration activity of each group was detected by methyl thiazol tetrazolium （MTT） assay. Enzyme reporter experiments verify the targeted regulation of SOX5 by miR - 23. Detection of cell proliferation activity after cotransfection of over - expression of SOX5 and miR - 23 mimic. Results The results of FQ - PCR showed that the expression level of miR -23 in miR -23 mimic group （0. 54 ±0. 13） was lower than that in NC group （ 1.03 ± 0.09, P = 0. 000） and control group （ 1.00, P = 0. 001 ） 48 h after transfection; proliferative activity compared with the other two groups, the proliferative activity of pituitary adenoma cells was decreased at 24, 48, 72 and 96 hours after transfection （A24h = 0. 47 ± 0. 04, A48h = 0. 52 ± 0. 05, A72h =0. 54 ± 0. 05, A96h = 0. 55 ± 0. 06, compared with the control group, P1 = 0. 036, P2 = 0. 019, P3 =0.006, P4 = 0. 000）. Luciferase reporter assays indicate that miR - 23 can target SOX5 mRNA. After transfection of PeDNA - SOXS, cell proliferation assay showed that overexpression of SOX5 could abolishthe mimicy effect of miR - 23 mimic on cell proliferation. Conclusion miR - 23 can regulate the expression of SOX5 and inhibit the proliferation of pituitary adenomas.

关 键 词：脑垂体腺瘤 微小RNA-23 SRY相关高迁移率族盒蛋白-5 细胞增殖 

分 类 号：R735.2[医药卫生—肿瘤]