摘要
目的探讨长链非编码RNA FAM66E通过微小RNA-544a(miR-544a)调控非小细胞肺癌(NSCLC)细胞增殖、迁移和侵袭的作用。方法采用实时荧光定量PCR(qPCR)检测NSCLC细胞(HCC827、NCI-H1975、A549和NCI-H23)及人正常肺上皮细胞BEAS-2B的FAM66E和miR-544a水平,双荧光素酶报告基因实验鉴定FAM66E和miR-544a间的关系。将NCI-H23细胞分为4组:对照组(未转染)、FAM66E过表达组(转染FAM66E过表达质粒pcDNA3.1-FAM66E+miR-544a NC)、miR-544a过表达组(转染pcDNA3.1空质粒+miR-544a模拟物mimics)和共表达组(转染pcDNA3.1-FAM66E+miR-544a mimics)。MTT实验、划痕实验和Transwell小室实验分别检测转染处理后细胞增殖、迁移和侵袭能力的变化。结果与BEAS-2B细胞相比,NSCLC细胞的FAM66E水平降低而miR-544a水平升高(P<0.05);miR-544a mimics显著降低了野生型FAM66E的荧光素酶活性(P<0.05),而对突变型FAM66E的不产生显著影响(P>0.05)。与对照组相比,FAM66E过表达组转染处理72 h后NCI-H23细胞活力减弱(P<0.05),miR-544a过表达组则增强(P<0.05),而共表达组的无明显改变(P>0.05)。FAM66E过表达组的穿膜细胞数较对照组的(181.833±9.135)个降低至(69.680±4.313)个,划痕愈合率较对照组的(46.337±3.424)%减少至(26.810±3.558)%,miR-544a过表达组的穿膜细胞数和划痕愈合率较对照组增多至(266.137±15.377)个和(69.392±2.551)%,上述差异均有统计学意义(P<0.05);共表达组的穿膜细胞数和划痕愈合率为(148.047±8.110)个和(48.460±4.143)%,与对照组的差异无统计学意义(P>0.05)。结论NSCLC细胞中FAM66E水平降低而miR-544a升高,FAM66E可能通过靶向miR-544a来抑制NCI-H23细胞的增殖、迁移和侵袭,在NSCLC中发挥抑瘤作用。
Objective To elucidate the potential influence of long non-coding RNA FAM66E on the proliferation,migration and invasion of non-small cell lung cancer(NSCLC)cells by regulating microRNA-544a(miR-544a).Methods Levels of FAM66E and miR-544a in human normal lung epithelial cell BEAS-2B and NSCLC cells including HCC827,NCI-H1975,A549 and NCI-H23 were detected by quantitative real-time PCR(qPCR).Dual luciferase gene reporter assay was used to identify the relationship between FAM66E and miR-544a.NCI-H23 cell were selected and allocated into four groups:control group(cells without transfection),FAM66E overexpression group(cells transfected with FAM66E overexpression plasmid pcDNA3.1-FAM66E and miR-544a NC),miR-544a overexpression group(cells transfected with pcDNA3.1 empty plasmid and miR-544a mimics)and co-expression group(cells transfected with pcDNA3.1-FAM66E and miR-544a mimics).MTT assay,scratch test and Transwell chamber test were used to detect the changes of cell proliferation,migration and invasion after transfection.Results Compared with BEAS-2B cells,the FAM66E levels decreased but miR-544a levels increased in NSCLC cells(P<0.05).MiR-544a mimics significantly decreased the relative luciferase activity of wild-type FAM66E(P<0.05),but had no significant effect on the relative luciferase activity of mutant FAM66E(P>0.05).Compared with the control group,the viability of NCI-H23 cells in FAM66E overexpression group was significantly decreased after 72 h of transfection(P<0.05),while that in miR-544a overexpression group was increased(P<0.05),but there was no significant change in co-expression group in relative to control group(P>0.05).The number of transmembrane cells in FAM66E overexpression group decreased to 69.680±4.313 compared with 181.833±9.135 of control group,and scratch healing rate decreased to(26.810±3.558)%compared with(46.337±3.424)%of control group(P<0.05).The number of transmembrane cells and scratch healing rate in miR-544a overexpression group were increased to 266.137±15.377 and(69.392±2
作者
宋晓燕
杨娟
袁青玲
SONG Xiaoyan;YANG Juan;YUAN Qingling(Department of Oncology, Qinghai Provincial People's Hospital, Xining 810001, China)
出处
《临床肿瘤学杂志》
CAS
2021年第4期302-307,共6页
Chinese Clinical Oncology
