转染miR-133a过表达对人食管癌EC9706细胞生长的影响及启膈散干预作用  

The effect of overexpression of transfected miR-133a on the growth of EC9706 cells and the intervention effect of Qige Powder

作　　者：李墨颜 王蕊 李丹丹[1] 刘陆 崔姗姗[1] 陈玉龙[1] 高小玲[1] LI Mo-yan;WANG Rui;LI Dan-dan;LIU Lu;CUI Shan-shan;CHEN Yu-long;GAO Xiao-ling(Henan University of Chinese Medicine,Zhengzhou Henan 450046,China)

机构地区：[1]河南中医药大学,河南郑州450046

出　　处：《时珍国医国药》2022年第4期837-841,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81803981);河南省高等学校青年骨干教师培养项目(2018GGJS084)。

摘　　要：目的优化miR-133a转染EC9706细胞条件,观察转染后启膈散对EC9706细胞增殖和凋亡的影响。方法使用荧光显微镜及流式细胞术优化EC9706细胞miR-133a转染条件;MTT法检测转染miR-133a后启膈散对EC9706细胞活性的抑制作用;流式细胞术检测转染miR-133a后启膈散对EC9706细胞的凋亡的影响。结果从荧光显微镜观察到的荧光信号强度及流式细胞仪检测荧光转染效率来看,24孔板EC9706细胞转染实验使用1L/孔Lipofectamine 2000(脂质体2000转染试剂)和100nmol·L^(-1)miR-133a时转染效果最佳;应用启膈散及转染miR-133a模拟物后对EC9706细胞活性有明显抑制作用(P<0.05),两者联合应用呈现一定的竞争抑制作用(P<0.01);EC9706细胞转染miR-133a模拟物48h后,与空白对照组相比,可增加细胞总凋亡率、细胞晚期凋亡率(P<0.01);启膈散作用于EC9706细胞48h后,与空白对照组相比可增加细胞总凋亡率、细胞早期凋亡率及细胞晚期凋亡率(P<0.01);两者联合应用对EC9706细胞的总凋亡率具有协同增效作用(P<0.01)。结论miR-133a过表达可促进EC9706细胞的增殖和凋亡,启膈散可通过促进miR-133a的表达影响EC9706细胞的生长。Objective To optimize the transfection conditions of miR-133 a into EC9706 cells,and observe the effect of Qige Powder on proliferation and apoptosis of EC9706 cells after transfection.Methods Optimization of transfection conditions of miR-133 a in EC9706 cells by fluorescence microscope and flow cytometry;MTT assay was used to detect the inhibitory effect of Qige Powder on EC9706 cells activity after transfection of miR-133 a;The effect of Qige Powder after transfection of miR-133 a on apoptosis of EC9706 cells was detected by flow cytometry.Results According to the fluorescence signal intensity observed by fluorescence microscope and the fluorescence transfection efficiency detected by flow cytometry,the transfection effect of EC9706 cells with 24-well plate was the best when using 1 L/well Lipofectamine 2000 and 100 nmol·L^(-1)miR-133 a;The application of Qige Oowder and transfection of miR-133 a mimic significantly inhibited the activity of EC9706 cells(P<0.01),and the combined application of Qige Powder and miR-133 a mimic showed a certain competitive inhibitory effect(P<0.01);Compared with the blank control group,EC9706 cells transfected with miR-133 a mimic for 48 hours can increase the total apoptosis rate and late apoptosis rate(P<0.01).Compared with the blank control group,Qige Powder can increase the total apoptosis rate,early apoptosis rate and late apoptosis rate of EC9706 cells after 48 hours of treatment(P<0.01);the combined application of the two has synergistic effect on the total apoptosis rate of EC9706 cells(P<0.01).Conclusion Overexpression of miR-133 a can promote the proliferation and apoptosis of EC9706 cells,and Qige Powder can influence the growth of EC9706 cells by promoting the expression of miR-133 a.

关 键 词：转染 miR-133a EC9706细胞 增殖 凋亡 

分 类 号：R285.5[医药卫生—中药学]