基础医学与临床 ›› 2019, Vol. 39 ›› Issue (1): 63-69.

• 研究论文 • 上一篇    下一篇

miR-145抑制人肝内胆管细胞癌细胞增殖

熊新魁,杜也牧,刘雨亭,顾殿华   

  1. 南京医科大学附属淮安第一医院
  • 收稿日期:2017-11-27 修回日期:2018-05-03 出版日期:2019-01-05 发布日期:2018-12-28
  • 通讯作者: 顾殿华 E-mail:docgudh@sina.com

miR-145 suppresses proliferation of human intrahepatic cholangiocarcinoma cells

  • Received:2017-11-27 Revised:2018-05-03 Online:2019-01-05 Published:2018-12-28

摘要: 目的 研究miR-145调节人肝内胆管细胞癌(ICC)细胞增殖的机制。方法 标本分为ICC癌组织和癌旁正常胆管组织,共38对,两种ICC细胞系:HuCCT-1和RBE,对其进行miR-145及NUAK1的表达调控。通过RT-qPCR检测miR-145和新式激酶家族1(NUAK1)的表达及两者相关性;检测细胞周期观察细胞增殖变化;荧光素酶报告基因实验确认miR-145的靶基因;Western blot观察AKT通路蛋白水平变化。结果 miR-145在癌组织的表达明显低于癌旁组织(p<0.05),NUAK1在癌组织的表达明显高于癌旁组织(p<0.05),两者具有相关性(p<0.05)。miR-145过表达组和NUAK1干扰组细胞计数均明显少于对照组(p<0.05);HuCCT-1细胞系转染miR-145 mimics后,G1期的细胞数明显增加(p<0.05);野生型的荧光信号明显被抑制(p<0.05);过表达miR-145或抑制NUAK1后,pAKT和pFOXO1表达明显下降(p<0.05)。结论 miR-145可能通过抑制NUAK1及其下游通路抑制ICC细胞增殖,它有可能为ICC临床诊治提供新的靶点和思路。

关键词: miR-145, 新式激酶家族1, 肝内胆管细胞癌, 蛋白激酶B

Abstract: Objective To investigate the specific mechanism of miR-145 regulating cell proliferation of human intrahepatic cholangiocarcinoma (ICC). Methods 38 pairs of ICC tissues and adjacent tissues were collected. At the same time, to investigate miR-145 function in ICC cells, HuCCT-1 and RBE ICC cell lines were transfected with miR-145 mimics and siRNA-mediated knockdown of NUAK1. The expression of miR-145 and NUAK1 was detected by RT-qPCR. And cell proliferation was observed by cell proliferation and cycle analysis. The target genes of miR-145 were confirmed by the luciferase report. The protein level of AKT pathway was studied by Western blot. Results The expressions of miR-145 in cancer tissues were significantly lower than that in adjacent tissues (p<0.05). While NUAK1 expressions in cancer tissues were significantly higher than that in adjacent tissues (p<0.05), and there was a certain correlation between them. The cell counts of the miR-145 overexpression group and the NUAK1 interference group were less compared with the negative control group (p<0.05). The number of cells in the G1 phase were increased markedly (p<0.05). Furthermore, the wild type fluorescence signal was suppressed significantly (p<0.05). The expression of pAKT and pFOXO1 was significantly decreased, while tAKT and tFOXO1 showed no significant change (p<0.05). Conclusion miR-145 may prevent ICC proliferation by targeting NUAK1 and its downstream effectors, and can therefore be useful for clinical diagnosis and targeted therapy of ICC.

Key words: miR-145, Novel (nua) kinase family (NUAK)1, intrahepatic cholangiocarcinoma, protein kinase B

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