miR-30a-5p/MTA1轴与喉鳞状细胞癌细胞Hep2生物学特性的相关性研究  

Study on the correlation between miR-30a-5p/MTA1 axis and the biological characteristics of LSCC cell Hep2

作　　者：裴婷 张海利[1] 陈英[1] 史聪 樊慧娟 Pei Ting;Zhang Haili;Chen Ying(Dept of Otolaryngology Head&Neck Surgery,First Hospital of Shanxi Medical University,Taiyuan 030001)

机构地区：[1]山西医科大学第一临床医院耳鼻咽喉头颈外科,太原030001

出　　处：《安徽医科大学学报》2021年第4期545-549,共5页Acta Universitatis Medicinalis Anhui

基　　金：山西省卫生和计划生育委员会科研基金(编号:2017036);山西省自然科学基金(编号:201901D111359)。

摘　　要：目的探讨miR-30a-5p通过靶向调控MTA1对喉鳞癌细胞增殖、迁移、周期和凋亡的影响。方法通过生物信息技术预测miR-30a-5p靶基因MTA1,并采用实时荧光定量PCR和双荧光素酶报告实验验证。将miR-30a-5p模拟物(mimics)和对照组(NC)分别转染至对数期的喉鳞癌细胞株Hep2中。利用CCK-8实验检测细胞增殖情况,Transwell实验检测细胞迁移情况,流式细胞术检测细胞周期和凋亡情况。结果实时荧光定量PCR实验结果显示,实验组和对照组中转染后表达水平分别为(12.62±0.97)和(1.04±0.32),差异有统计学意义(P<0.05)。双荧光素酶报告实验结果显示,miR-30a-5p模拟物+pGL4.10和+pGL 4.73分别与其野生型和突变型比较表达水平为(0.41±0.06)和(0.91±0.01),差异有统计学意义(P<0.05)。在CCK-8实验结果显示,实验组比对照组24、48、72 h的吸光度值相对倍数低,差异有统计学意义(P<0.05)。Transwell细胞迁移实验结果显示,实验组和对照组细胞数目分别为(207.67±7.23)个和(369.00±41.22)个,差异有统计学意义(P<0.05)。在流式细胞术检测细胞周期结果示,实验组G1期、S期和G2期均较miR-NC对照组小,细胞周期阻滞;细胞凋亡结果示,实验组和对照组细胞凋亡率分别为(22.61±5.70)%和(4.52±2.12)%,差异有统计学意义(P<0.05)。结论miR-30a-5p/MTA1轴可能对喉鳞状细胞癌的发生机制产生一定的阻碍作用。Objective To investigate the effects of miR-30a-5p on the proliferation,migration,cycle and apoptosis of laryngeal squamous cell carcinoma cells by targeting the regulation of MTA1.Methods The target gene MTA1 of miR-30a-5p was predictedby biological information technology and verified by RT-qPCR and dual-luciferase reporting experiment.miR-30a-5p mimics and NC were transfected into LSCC cell line Hep2 at logarithmic phase.Cell proliferation was detected by CCK-8 assay.Transwell assay was used to detect cell migration.Flow cytometry was used to detect cell cycle and apoptosis.Results In the real-time quantitative PCR results,the post-transfection expression levels of experimental group and control group were(12.62±0.97)and(1.04±0.32),respectively,with statistically significant difference(P<0.05).In the results of the double luciferase report experiment,the experiment levels of miR-30a-5p mimics+pGL4.10+pGL 4.73 were(0.41±0.06)and(0.91±0.01),respectively,when compared with the wild-type and mutant-type,and the differencewas statistically significant(P<0.05).In the results of CCK-8 experiment,the absorbance value of the experimental group was lower than of the control group at 24,48,72 h,and the difference was statistically significant(P<0.05).In the Transwell cell migration experiment,the number of the experimental group and the control group was(207.67±7.23)and(369.00±41.22),respectively,with statistically significant differences(P<0.05).The results of flow cytometry detection of cell cycle showed that the G1,S and G2 phases of the experimental group were smaller than those of the control group,with cell cycle arrest.The results of cell apoptosis showed that the apoptosis rates of the experimental group and the control group were(22.61±5.70)%and(4.52±2.12)%respectively,with statistically significant differences(P<0.05).Conclusion The miR-30a-5p/MTA1 axis may have a certain inhibitory effect on the pathogenesis of laryngeal squamous cell carcinoma.

关 键 词：喉鳞状细胞癌 miR-30a-5p MTA1 增殖 迁移 

分 类 号：R767.19[医药卫生—耳鼻咽喉科]