摘要
目的检测微小RNA(miRNA)-153和ATP结合盒E1(ABCE1)在肺鳞状细胞癌(LSCC)组织和细胞中的相对表达水平,并探究其对LSCC细胞侵袭的影响。方法收集65例LSCC患者的肿瘤组织和癌旁组织。应用定量反转录PCR检测LSCC组织和细胞中miRNA-153和ABCE1 mRNA相对表达水平,应用双荧光素酶报告基因实验分析LSCC细胞中miRNA-153与ABCE1的靶向关系。将ABCE1干扰质粒(si-ABCE1)转染的LSCC细胞(SK-MES-1、NCI-H520)设为si-ABCE1组,同时设置si-NC组(阴性对照组)和对照组(空白对照组),应用定量反转录PCR和蛋白质印迹法实验共同检测转染效果。应用3-(4,5-二甲基-2-噻唑)-2,5-二苯基四氮唑溴盐实验和Transwell实验分别检测敲低ABCE1对细胞增殖和侵袭的影响。结果LSCC组织中miRNA-153 mRNA的相对表达水平低于LSCC癌旁组织,LSCC组织中ABCE1 mRNA相对表达水平高于LSCC癌旁组织;LSCC细胞SK-MES-1、NCI-H520中miRNA-153 mRNA的相对表达水平均低于正常人细胞BEAS-2B,LSCC细胞SK-MES-1、NCI-H520中ABCE1 mRNA相对表达水平均高于正常人细胞BEAS-2B(均P<0.05)。双荧光素酶报告基因实验结果显示,ABCE1 WT+miRNA-153 mimic细胞的相对荧光素酶活性低于ABCE1 WT细胞(P<0.05)。在LSCC细胞SK-MES-1和NCI-H520中,si-ABCE1组光密度值和穿膜细胞数均少于对照组和si-NC组(均P>0.05)。结论miRNA-153在LSCC中表达降低,ABCE1在LSCC中表达升高,miRNA-153可能通过靶向结合ABCE1抑制LSCC细胞侵袭。
Objective To detect the relative expression levels of microRNA(miRNA)-153 and ATP-binding cassette E1(ABCE1)in lung squamous cell carcinoma(LSCC)tissue and cell,and to explore their effects on LSCC cell invasion.Method Tumor tissue and paracancerous tissue of 65 LSCC patients were collected.Quantitative reverse transcription PCR was used to detect the relative expression levels of miRNA-153 and ABCE1 mRNAs in LSCC tissue and cell,and the dual-luciferase reporter gene assay was used to analyze the targeting relationship between miRNA-153 and ABCE1 in LSCC cell.LSCC cells(SK-MES-1,NCI-H520)transfected with ABCE1 interference plasmid(si-ABCE1)were set as the si-ABCE1 group,the si-NC group(negative control group)and the control group(blank control group)were set up at the same time,and the quantitative reverse transcription PCR and western blotting were used to detect the transfection effect.The effects of ABCE1 knockdown on cell proliferation and invasion were respectively detected by 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide assay and Transwell experiment.Results The relative expression level of miRNA-153 mRNA in LSCC tissue was lower than that in LSCC paracancerous tissue,while the relative expression level of ABCE1 mRNA in LSCC tissue was higher than that in LSCC paracancerous tissue;the relative expression levels of miRNA-153 mRNA in LSCC cells(SK-MES-1 and NCI-H520)were lower than that in healthy people cell(BEAS-2B),while the relative expression levels of ABCE1 mRNA in LSCC cells(SK-MES-1 and NCI-H520)were lower than that in healthy people cell(BEAS-2B)(all P<0.05).The results of the dual-luciferase reporter gene assay showed that the relative luciferase activity of the ABCE1 WT+miRNA-153 mimic cell was lower than that of the ABCE1 WT cell(P<0.05).In LSCC cells(SK-MES-1 and NCI-H520),the optical density values and the numbers of transmembrane cells in the si-ABCE1 group were lower than those in the control group and the si-NC group(all P>0.05).Conclusion The expression of miRNA-153 is decreased in LSCC,while the expression of ABCE1 is elevated in LSCC.MiRNA-153 may inhibit LSCC cell invasion by targeting ABCE1.
作者
杨阳
孙艳丽
郭秋杰
YANG Yang;SUN Yanli;GUO Qiujie(Department of Laboratory Medicine,Xuchang Hospital of Traditional Chinese Medicine,Xuchang 461000,Henan Province,China)
出处
《内科》
2023年第3期247-252,共6页
Internal Medicine