摘要
目的:研究微小RNA-218(microRNA-218,miR-218)靶向B细胞特性莫洛尼鼠白血病病毒插入位点1(B-cell specific moloney leukemia virus insertion site 1,Bmi-1)抑制肺腺癌细胞增殖的作用及机制。方法:培养肺腺癌细胞株A549,分别转染miR-阴性对照(NC)核苷酸序列、miR-218核苷酸序列、NC质粒、Bmi-1质粒,采用平板克隆形成实验、CCK8实验检测细胞增殖水平,采用荧光定量PCR实验检测miR-218的表达,采用Western blot实验检测Bmi-1的表达,采用双荧光素酶报告基因实验检测miR-218与Bmi-1的靶向关系。建立移植瘤裸鼠,慢病毒转染miR-NC或miR-218核苷酸序列,检测移植瘤体积、质量及Bmi-1的表达。结果:与转染miR-NC核苷酸序列的miR-NC组比较,转染miR-218核苷酸序列的miR-218组A549细胞miR-218表达水平、增殖抑制率增加,克隆形成率、Bmi-1表达水平及野生型Bmi-1质粒的荧光素酶活性降低(P<0.05);与共转染miR-218核苷酸序列及NC质粒的miR-218+NC质粒组比较,共转染miR-218核苷酸序列及Bmi-1质粒的miR-218+Bmi-1质粒组A549细胞增殖抑制率降低,克隆形成率、Bmi-1表达水平增加(P<0.05);与慢病毒转染miR-NC核苷酸序列的miR-NC组比较,慢病毒转染miR-218核苷酸序列的miR-218组裸鼠移植瘤体积、质量及Bmi-1表达水平均降低(P<0.05)。结论:miR-218靶向下调Bmi-1表达并抑制肺腺癌细胞增殖。
Objective:To investigate the inhibitory effect and mechanism of B-cell specific moloney leukemia virus insertion site 1(Bmi-1)on the proliferation of lung adenocarcinoma cells by targeting microRNA-218(miR-218).Methods:The lung adenocarcinoma cell line A549 was cultured and transfected with miR-negative control(NC)nucleotide sequence,miR-218 nucleotide sequence,NC plasmid and Bmi-1 plasmid,respectively.The cell proliferation level was detected by plate cloning test and CCK8 test,the expression of miR-218 was detected by fluorescence quantitative PCR,the expression of Bmi-1 was detected by Western blot,and the targeting relationship between miR-218 and Bmi-1 was detected by double luciferase reporter gene.Nude mice with transplanted tumor was established,lentivirus was transfected with miR-NC or miR-218 nucleotide sequence and then the volume,quality and Bmi-1 expression of transplanted tumor was detected.Results:Compared with the miR-NC group transfected with miR-NC nucleotide sequence,the miR-218 expression level and proliferation inhibition rate of A549 cells increased,and the cloning rate,Bmi-1 expression level and luciferase activity of wild type Bmi-1 plasmid decreasedin the miR-218 group transfected with miR-218 nucleotide sequence(P<0.05).Compared with the miR-218+NC plasmid group co-transfected with miR-218 nucleotide sequence and NC plasmid,the inhibition rate of A549 cell proliferation decreased,while the clone formation rate and Bmi-1 expression level increased in the miR-218+Bmi-1 plasmid group co-transfected with miR-218 nucleotide sequence and Bmi-1 plasmid(P<0.05).Compared with the miR-NC group transfected with miR-NC nucleotide sequence of lentivirus,the volume,quality and Bmi-1 expression level of transplanted tumor in nude mice of miR-218 group transfected with miR-218 nucleotide sequence of lentivirus decreased(P<0.05).Conclusion:miR-218 suppresses the proliferation of lung adenocarcinoma cell by targeted downregulation of Bmi-1 expression.
作者
焦会珍
金小乐
苑萌
王天伦
JIAO Huizhen;JIN Xiaole;YUAN Meng;WANG Tianlun(Department of Emergency,the First Affiliated Hospital of Hebei North University,Hebei Zhangjiakou 075000,China)
出处
《现代肿瘤医学》
CAS
北大核心
2023年第23期4301-4306,共6页
Journal of Modern Oncology
基金
河北省2023年度医学科学研究课题(编号:20231456)。
关键词
肺腺癌
MIR-218
BMI-1
靶基因
增殖
移植瘤
lung adenocarcinoma
miR-218
Bmi-1
target genes
proliferation
transplant tumor Modern Oncology 2023
31(23):43
