微小核糖核酸-589-5p对喉癌细胞增殖和凋亡的影响及其机制  

Effects of micro ribonucleic acid 589-5p on proliferation and apoptosis of laryngeal cancer cells and underlying mechanism

作　　者：罗敏[1] 潘春晨 孙刚[1] 段金芳[1] 陈红[1] 陈天红 Luo Min;Pan Chunchen;Sun Gang;Duan Jinfang;Chen Hong;Chen Tianhong(Department of Otolaryngology and Head and Neck Surgery,Chaohu Hospital Affiliated to Anhui Medical University,Hefei 238000,China;Department of Otolaryngology and Head and Neck Surgery,the First Affiliated Hospital of University of Science and Technology of China,Hefei 230001,China)

机构地区：[1]安徽医科大学附属巢湖医院耳鼻咽喉头颈外科,合肥238000 [2]中国科学技术大学附属第一医院耳鼻咽喉头颈外科,合肥230001

出　　处：《中华实验外科杂志》2022年第2期332-335,共4页Chinese Journal of Experimental Surgery

基　　金：2021年安徽医科大学校科研基金立项资助项目(2021xkj187)。

摘　　要：目的探讨微小核糖核酸-589-5p(miR-589-5p)对喉癌细胞增殖和凋亡的影响及其机制。方法喉癌细胞系(TU177、TU686和AMC-HN-8)及人正常支气管上皮细胞(16HBE)购自美国典型培养物保藏中心。将miR-NC、miR-589-5p、si-NC和si-FGD4转染至TU686细胞(分别标记为miR-NC组、miR-589-5p组、si-NC组和si-FGD4组);miR-589-5p与pcDNA、miR-589-5p与pcDNA-FGD4共转染至TU686细胞(分别标记为miR-589-5p+pcDNA组和miR-589-5p+pcDNA-FGD4组)。溴化-3-(4, 5-二甲基-2-噻唑)-2, 5-二苯基四氮唑(MTT)检测细胞增殖;流式细胞仪检测细胞凋亡;实时荧光定量聚合酶链反应(qRT-PCR)检测miR-589-5p和FYVE、RhoGEF和PH域包含4(FYVE, RhoGEF And PH domain containing 4, FGD4)基因信使核糖核酸(mRNA)表达水平;蛋白质印迹法检测FGD4、细胞周期蛋白D1(Cyclin D1)、B细胞淋巴瘤-2(bcl-2)、p21、B细胞淋巴瘤-2相关X蛋白(bax)蛋白表达。两组间比较行t检验, 多组间比较采用单因素方差分析, 组间两两比较采用SNK检验。结果 miR-589-5p组喉癌细胞凋亡率(31.77±3.26)%比(18.82±1.27)%、miR-589-5p(0.74±0.06比0.31±0.02)、p21(0.78±0.05比0.37±0.02)和bax(0.68±0.04比0.31±0.03)表达水平显著高于miR-NC组, 细胞活性(0.47±0.03比0.79±0.06)、Cyclin D1(0.25±0.02比0.66±0.04)及bcl-2(0.18±0.02比0.59±0.03)表达水平显著低于miR-NC组, 差异有统计学意义(t=18.891、8.276、12.452、21.439、11.766、19.231、10.545, P<0.05)。转染miR-589-5p组喉癌细胞FGD4基因mRNA(0.41±0.03比0.72±0.05)及蛋白表达(0.25±0.02比0.45±0.03)显著低于miR-NC组细胞, 差异有统计学意义(t=21.133、15.509, P<0.05);转染anti-miR-589-5p组喉癌细胞FGD4基因mRNA(0.81±0.07比0.56±0.05)及蛋白表达(0.51±0.04比0.32±0.03)显著高于anti-miR-NC组细胞, 差异有统计学意义(t=15.024、20.235, P<0.05)。miR-589-5p+pcDNA-FGD4组细胞活性(0.79±0.06比0.62±0.04)、FGD4(0.68±0.04比0.31±0.03)、Cyclin D1(0.66±0.04比0.28±0.03)和bcl-2(0.61±0.04比0.34±0.Objective To study the effects and mechanism of micro ribonucleic acid-589-5p(miR-589-5p)on proliferation and apoptosis of laryngeal cancer cells.Methods The miR-NC,miR-589-5p,si-NC and si-FGD4 were transfected into TU686 cells(marked as miR-NC group,miR-589-5p group,si-NC group and si-FGD4 group,respectively).The miR-589-5p and pcDNA,and miR-589-5p and pcDNA-FGD4 were co-transfected into TU686 cells(marked as miR-589-5p+pcDNA group and miR-589-5p+pcDNA-FGD4 group,respectively).Bromide-3-(4,5-Dimethyl-2-thiazole)-2,5-diphenyltetrazolium Bromide,MTT assay was used to detect cell proliferation.The flow cytometry was used to detect cell apoptosis.The real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the mRNA expression of miR-589-5p and FYVE,RhoGEF and PH domains contain 4(FGD4)genes.Western blotting was used to detect the expression of FGD4,cyclin D1(Cyclin D1),B cell lymphoma-2(bcl-2),p21,bcl-2 associated X protein(bax)proteins.The t test was used for comparison between two groups,the one-way analysis of variance was used for comparison between multiple groups,and the SNK test was used for pairwise comparison between groups.Results The apoptosis rate in miR-589-5p group[(31.77±3.26)%vs.(18.82±1.27)%],miR-589-5p(0.74±0.06 vs.0.31±0.02),p21(0.78±0.05 vs.0.37±0.02)and bax(0.68±0.04 vs.0.31±0.03)were significantly higher than those in miR-NC group,while cell viability(0.47±0.03 vs.0.79±0.06),Cyclin D1(0.25±0.02 vs.0.66±0.04)and bcl-2(0.18±0.06 vs.0.59±0.03)were significantly lower(t=18.891,8.276,12.452,21.439,11.766,19.231,10.545,P<0.05).The cell viability in miR-589-5p+pcDNA-FGD4 group(0.79±0.06 vs.0.62±0.04),FGD4(0.68±0.04 vs.0.31±0.03),Cyclin D1(0.66±0.04 vs.0.28±0.03)and bcl-2(0.61±0.04 vs.0.34±0.03)were significantly higher than in miR-589-5p+pcDNA group,while the apoptosis rate[(20.17±2.44)%vs.(33.14±3.02)%],p21(0.41±0.03 vs.0.67±0.5)and bax(0.38±0.06 vs.0.59±0.04)were significantly lower(t=9.465,15.133,12.307,11.731,18.235,15.133,21.347,P<0.05).

关 键 词：喉癌 增殖 凋亡 

分 类 号：R739.65[医药卫生—肿瘤]