摘要
该研究主要探讨了微小核酸mi RNA-193a-5p对人胰腺癌细胞体外迁移和侵袭的促进作用及其机制。采用miR-193a-5p模拟物及mi R-193a-5p抑制剂分别上调和下调miR-193a-5p的表达;采用细胞划痕法和Transwell小室法检测mi R-193a-5p对细胞迁移和侵袭能力的影响;采用TargetScan 7.1数据库预测miR-193a-5p的靶基因;荧光素酶报告法验证miR-193a-5p的靶基因;Western blot和实时荧光定量PCR验证其表达结果。结果表明, miR-193a-5p可显著促进细胞的迁移和侵袭能力。TargetScan 7.1软件预测, Prox1可能为mi R-193a-5p的靶基因,荧光素酶报告实验显示, miR-193a-5p靶向Prox1基因的3′UTR区。实时荧光定量PCR和Western blot结果显示, miR-193a-5p下调了Prox1的mRNA和蛋白水平的表达。研究结果揭示,在胰腺癌中高表达的miR-193a-5p通过下调Prox1,从而使胰腺癌细胞获得高的迁移和侵袭能力,促进胰腺癌的转移。
The aim of this study was to investigate the effect and mechanisms of miR-193 a-5 p on migration and invasion of human breast cancer cells in vitro. Panc-1 cells were treated with miR-193 a-5 p mimic or miR-193 a-5 p inhibitor to up-regulate/down-regulate the expression level of miR-193 a-5 p. Wound-healing assay and Transwell chamber were employed to determine cell migration and invasion in vitro. The target gene of miR-193 a-5 p was predicted with the TargetScan 7.1 database and verified through luciferase reporter method. The effects of miR-193 a-5 p on the expression of the potential target were detected by Real-time PCR and Western blot. Results showed that in vitro migration and invasion ability of Panc-1 cells was increased significantly by Real-time PCR, which could target Prox1 in the 3′UTR region. The mRNA and protein expression of Prox1 was decreased by miR-193 a-5 p overexpression. The results validated that miR-193 a-5 p, highly expressed in pancreatic cancer, could down-regulate Prox1, which in turn promotes migration and invasion ability of breast cancer cells, thus promoting pancreatic cancer metastasis.
作者
李曼曼
杨召聪
卢洲
潘怡
金亮
Li Manman;Yang Zhaocong;Lu Zhou;Pan Yi;Jin Liang(School of life Science and Technology,China Pharmaceutical University,Nanjing 210009,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2019年第8期1534-1542,共9页
Chinese Journal of Cell Biology
基金
国家自然科学基金(批准号:81570696、81702941)
江苏高校“青蓝工程”资助项目
高等学校学科创新引智计划(“111计划”)(批准号:B16046)
江苏高校优势学科建设工程资助项目(批准号:PAPD)资助的课题~~
