上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

Oct4和Nanog基因沉默对胰腺癌干细胞耐药性的影响

陆 洋1,2，陆玉华3，朱 慧3，单海燕3，陆俊杰3，钱海鑫1，王志伟3   

  1. 1.苏州大学附属第一医院普外科， 苏州 215006； 2.南通大学附属医院重症医学科， 南通 226001； 3.南通大学附属医院普外科， 南通 226001
  • 出版日期:2014-03-28 发布日期:2014-04-02
  • 通讯作者: 钱海鑫, 电子信箱: qianhaixin1@hotmail.com; 王志伟, 电子信箱: wzw3636@sina.com。
  • 作者简介:陆 洋（1978—）， 男， 主治医师， 硕士; 电子信箱: marineluyang@hotmail.com。
  • 基金资助:

    国家自然科学基金（81101615）; 江苏省自然科学基金（BK2010276）

Effect of Oct4 and Nanog gene silencing on drug resistance of pancreatic cancer stem cells

LU Yang1,2, LU Yu-hua3, ZHU Hui3, SHAN Hai-yan3, LU Jun-jie3, QIAN Hai-xin1, WANG Zhi-wei3   

  1. 1.Department of General Surgery， the First Hospital Affiliated to Soochow University， Suzhou 215006， China; 2.Intensive Care Unit, Affiliated Hospital of Nantong University, Nantong 226001, China; 3.Department of General Surgery, Affiliated Hospital of Nantong University, Nantong 226001, China
  • Online:2014-03-28 Published:2014-04-02
  • Supported by:

    National Natural Science Foundation of China，81101615；Natural Science Foundation of Jiangsu Province，BK2010276

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摘要：

目的 观察通过RNA干扰同时沉默Oct4和Nanog基因对胰腺癌干细胞（PCSCs）化疗药物耐药性的影响，探索提高胰腺癌化疗效果的新方法。方法 通过流式细胞仪从人胰腺癌细胞系PANC-1中分离出CD44+CD24+ESA+的PCSCs，体外建株，通过构建特异性慢病毒shRNA-Oct4/Nanog载体同时沉默PCSCs的Oct4和Nanog基因，分选出基因沉默的PCSCs （PCSCs-shOct4+shNanog），通过CCK8细胞活性检测法观察其对化疗药物吉西他滨敏感性的变化，Real-Time RT-PCR和Western blotting检测耐药相关基因ABCG2的表达。结果 CD44+CD24+ESA+的PCSCs高度表达Oct4和Nanog基因，在体外表现为对吉西他滨的耐药性增强。Oct4和Nanog基因同时被沉默后PCSCs干性特征出现抑制，ABCG2表达下降，耐药性降低，差异均有统计学意义（P<0.05）。结论 Oct4和Nanog基因在保持PCSCs肿瘤干性特性方面发挥重要作用，沉默二者表达后PCSCs的耐药性受到抑制。

关键词: Oct4, Nanog, 胰腺癌干细胞, 基因沉默

Abstract:

Objective To investigate the effects of Oct4 and Nanog gene silenced by shRNA on drug resistance of pancreatic cancer stem cells and explore new ways of improving chemotherapeutic effect on the pancreatic cancer. Methods The CD44+CD24+ESA+ pancreatic cancer stem cells (PCSCs) were isolated from PANC-1 cell line by flow cytometry and constructed cell strains in vitro. We constructed lentiviral virus-shRNA-Oct4/Nanog vector target Oct4 and Nanog and silenced them simultaneously, and then identified whether PCSCs inhabited expressions of the two genes (PCSCs-shOct4+shNanog) showed the changes of drug resistance to gemcitabine by cell counting Kit-8 (CCK-8) cell proliferation assay and the expression of ABCG2 were examined by Real-Time RT-PCR and Western blotting analysis. Results The high expressions of Oct4 and Nanog were in CD44+CD24+ESA+ PCSCs, which indicated enhanced resistance to gemcitabine in vitro. Double knockdown of Oct4 and Nanog significantly reduced stemness of PCSCs, expression of ABCG2, and chemoresistance to gemcitabine. The differences were statistically significant (P<0.05). Conclusion The results suggest that Oct4 and Nanog play critical roles in maintaining the stemness of PCSCs. Gene silencing of Oct4 and Nanog may inhabit the drug resistance of PCSCs.

Key words: Oct4, Nanog, pancreatic cancer stem cells, gene silencing