基础医学与临床 ›› 2018, Vol. 38 ›› Issue (8): 1135-1140.

• 研究论文 • 上一篇    下一篇

miR-145对胰腺癌细胞系PANC-1上皮间质转化及肿瘤干细胞增殖的影响

邓大炜1,吴斌2,严舒2,兰川1,张光年1,曾丽娟1,李建水1   

  1. 1. 川北医学院附属医院
    2. 川北医学院附属医院 肝胆外科
  • 收稿日期:2018-04-28 修回日期:2018-06-22 出版日期:2018-08-05 发布日期:2018-07-24
  • 通讯作者: 李建水 E-mail:q422528137@qq.com
  • 基金资助:
    超声靶向微泡联合NF-κB核定位基序介导mir-145转染影响胰腺癌肿干细胞特性获得

Effect of miR-145 on EMT and proliferation of cancer stem cell in pancreatic cancer cell line PANC-1

  • Received:2018-04-28 Revised:2018-06-22 Online:2018-08-05 Published:2018-07-24

摘要: 目的 探讨miR-145对胰腺癌细胞系PANC-1发生上皮间质转化(EMT)及肿瘤干细胞特性的影响及相关机制。方法 设miR-145组、对照组与空白组;构建miR-145过表达慢病毒,转染到PANC-1细胞中;用RT-qPCR法检测各组miR-145的表达;用黏附试验检测细胞黏附能力的变化,用瘤球形成实验检测胰腺癌干细胞形成的影响;用RT-qPCR和Western blot检测E-cadherin、N-cadherin与OCT4的mRNA和蛋白质的差异。结果 转染miR-145过表达慢病毒后能够明显增加miR-145的mRNA水平(p<0.05);miR-145组相对对照组与空白组PANC-1细胞同种黏附能力增加、异种黏附能力降低、抑制肿瘤干细胞球数量及直径(p<0.05);miR-145组相对对照组与空白组,E-cadherin表达明显增高而N-cadherin和OCT4在mRNA与蛋白质表达均明显降低(p<0.05)。结论 上调miR-145能够促进胰腺癌细胞系PANC-1 EMT的逆转,降低胰腺癌肿瘤干细胞特性获得,其机制可能其机制可能与E-cadherin的上调及N-cadherin和OCT4的下调有关。

关键词: 胰腺癌, 肿瘤干细胞, 上皮间质转化(EMT), miR-145

Abstract: Objective To investigate the effect of miR-145 on EMT and proliferation of pancreatic cancer cell line PANC-1 and to explore the related mechanism. Methods miR-145 group, contral group and blank group were set up.miR-145 over-expressed lentiviral vector was constructed and transfected into PANC-1 cells. RT-qPCR was used to testify miR-145 mRNA expressions;adhesion experiments were used to detect the ability of homogeneous and heterogeneous cells intercellular adhesion of each group, which was observed under fluorescence microscopy.The characteristic changes of tumor stem cells were measured by spheroid formation assay; Meanwhile, RT-qPCR and Western blotting were performed to examine mRNA and protein expressions of E-cadherin、N-cadherin and OCT4, respectively. Results The expression of mRNA of miR-145 significantly was increased after transfecting miR-145 over-expressed lentiviral vector; Compared with contral group and blank group,the miR-145 group homogeneous cells intercellular adhesion was increased and heterogeneous cells intercellular was opposite,the number and the size of tumorspheres was obviously reduced(p<0.05). meanwhile,the expression of mRNA and protein of E-cadherin was up-regulated,and N-cadherin and OCT4 were declined(p<0.05). Conclusions Up-regulating miR-145 can reverse the EMT and inhibit the characteristic acquisition of cancer stem cell line PANC-1, its mechanism may be related to the up-regulation of E-cadherin and the down-regulation of N-cadherin and OCT4.

Key words: Pancreatic cancer, Cancer stem cells, epithelial-mesenchymal transition(EMT), MiRNA-145

中图分类号: