%0 Journal Article
%T KLF4转录水平负调控miR-106a表达对人胃癌细胞BGC-823侵袭能力的影响<br>Effect of miR-106a expression negatively regulated by KLF4 on the invasive activity of human gastric cancer cells BGC-823
%A 朱 萌
%A 张 宁
%A 和水祥
%A 张 丹
%A 张志勇
%J 西安交通大学学报(医学版)
%D 2019
%R 10.7652/jdyxb201903004
%X 摘要：目的 探讨转录因子kruppel样因子4(Krüppel-like factor 4, KLF4)对微小RNA-106a(miR-106a)表达的调控及其相互作用对人胃癌细胞侵袭能力的影响。方法 通过转录因子结合位点数据库JASPAR检索与miR-106a启动子区结合的转录因子KLF4。构建KLF4过表达载体(KLF4-pcDNA)和miR-106a报告基因(pmiR-106a-WT/MUT-luc)，双荧光素酶报告基因检测KLF4对miR-106a启动子活性的影响。收集人胃癌和癌旁石蜡包埋-甲醛固定(formalin-fixed paraffin-embedded, FFPE)标本各40例，实时定量PCR和免疫组织化学法检测KLF4表达。人胃癌细胞株BGC-823培养并按miR-106a mimic组、mimic NC组、KLF4+pcDNA组、pcDNA basic组分别进行处理，Transwell法检测各组胃癌细胞侵袭能力变化。 结果 双荧光素酶报告基因检测显示KLF4-pcDNA具有拮抗miR-106a启动子活性的作用，表现为pmiR-106a-WT-luc荧光素酶活性被抑制(pmiR-106a-WT+pcDNA-basic与pmiR-106a-WT+pcDNA-KLF4比较，P<0.001)，但对pmiR-106a-MUT-luc荧光素酶活性影响不明显(pmiR-106a-MUT+pcDNA-basic与pmiR-106a-MUT+pcDNA-KLF4比较，P>0.05)。FFPE样本显示KLF4在胃癌组织中的相对表达量为0.69±0.59，与癌旁组织相比，差异有统计学意义(P<0.001)，且与胃癌的细胞分化程度、淋巴转移和浸润深度相关(P<0.05)；KLF4阳性表达主要位于胃黏膜上皮细胞核及胞质。Transwell实验表明各处理组胃癌细胞的穿膜数量不全相同(P<0.001)，miR-106a mimic组为89.00±14.85，mimic NC组为50.90±17.94，KLF4+pcDNA组为37.70±12.60，pcDNA basic组为66.20±3.19。结论 转录因子KLF4与miR-106a启动子区至少存在体外的直接结合，可能通过在上游转录水平负调控miR-106a表达而参与影响胃癌细胞的侵袭转移进程。<br>ABSTRACT: Objective To investigate the effect of Krüppel-like factor 4 (KLF4) on the expression of miR-106a and their interaction on the invasive activity of human gastric cancer cells. Methods The JASPAR database was used to screen the transcriptional factor combined with miR-106a promoter. KLF4 over-expression vector KLF4-pcDNA and miR-106a reporter gene pmiR-106a-WT/MUT-luc were both constructed, and the dual luciferase reporter assay was used to detect the effect of KLF4 on the activity of miR-106a promoter. Forty specimens of human gastric cancer and their adjacent formalin-fixed paraffin-embedded (FFPE) specimens were collected, and Real-time PCR and immunohistochemistry were both used to detect the expression of KLF4. Human gastric cancer cell line BGC-823 was divided into four groups: miR-106a mimic, mimic NC, KLF4+pcDNA, and pcDNA basic. Transwell assay was employed to detect the invasive ability of the gastric cancer cells after four different treatments. Results Dual luciferase reporter assay indicated that KLF4-pcDNA could antagonize the activity of miR-106a promoter, which suggested that the activity of pmiR-106a-WT-luc luciferase was inhibited (pmiR-106a-WT+pcDNA-basic compared with pmiR-106a-WT+pcDNA-KLF4, P<0.001), but the pmiR-106a-MUT-luc luciferase was not significantly influenced (pmiR-106a-MUT+pcDNA-basic compared with pmiR-106a-MUT+pcDNA-KLF4, P>0.05). FFPE samples showed that the relative expression of KLF4 in gastric cancer was 0.69±0.59, which significantly differed from that in adjacent paracancerous tissues (P<0.001). The expression of KLF4 was correlated
%K 胃癌
%K Krüppel样因子4(KLF4)
%K 微小RNA-106a(miR-106a)
%K 转录因子
%K 启动子
%K 侵袭<br>gastric cancer
%K Krüppel-like factor 4
%K microRNA-106a
%K transcriptional factor
%K promoter
%K invasion
%U http://yxxb.xjtu.edu.cn//oa/darticle.aspx?type=view&id=201903004