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山东大学学报(医学版) ›› 2010, Vol. 48 ›› Issue (6): 80-83.

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RNA干扰技术沉默MDR1基因逆转大肠癌耐药细胞株LoVo/5-Fu的耐药性

孔帅,李乐平,靖昌庆,王峰   

  1. 山东大学附属省立医院胃肠外科, 济南 250021
  • 收稿日期:2010-01-26 出版日期:2010-06-16 发布日期:2010-06-16
  • 通讯作者: 靖昌庆(1974- ),男,博士,副主任医师,主要从事胃肠道肿瘤的临床治疗和基础研究。 E-mail: jing66510122@sina.com
  • 作者简介:孔帅(1984- ),男,硕士研究生,主要从事胃肠道肿瘤研究。
  • 基金资助:

    山东省医药卫生科技发展计划(2007-33)

The reversal effect on MDR1 gene-mediated multidrug resistance in human colon carcinoma LoVo/5-Fu cells by RNA interference

KONG Shuai, LI Leping, JING Changqing, WANG Feng   

  1. Department of Gastrointestinal Surgery,  Provincial Hospital Affiliated to Shandong University, Jinan 250021, China
  • Received:2010-01-26 Online:2010-06-16 Published:2010-06-16

摘要:

目的  探讨多药耐药(MDR1)基因沉默对大肠癌耐药细胞系LoVo/5-Fu耐药性的逆转作用。方法  构建靶向MDR1的短发夹RNA(EASY-shRNA-MDR1)干扰质粒转染LoVo/5Fu,实时荧光定量PCR(Realtime-PCR)在mRNA水平检测抑制效果,噻唑蓝(MTT)法检测细胞对5-Fu的敏感性,流式细胞仪检测细胞凋亡情况。结果  与未转染组相比,EASY-shRNA-MDR1组细胞MDR1mRNA表达明显下调(P<0.05);细胞对5-Fu的IC50及RI显著降低(P<0.05),敏感性的相对逆转率为74.7%;凋亡率明显升高(P<0.05)。结论  EASY-shRNA-MDR1可有效抑制大肠癌耐药细胞中MDR1的表达,进而降低了大肠癌耐药株对5-Fu的耐药性。

关键词: 结肠肿瘤;RNA干扰;基因,MDR1;细胞凋亡;基因治疗

Abstract:

Objective  To investigate the reversal effect on MDR1 gene-mediated multidrug resistance in human colon carcinoma LoVo/5-Fu cells by RNA interference. Methods  A eukaryotic expression plasmid of shRNA targeting on MDR1 was transiently transfected into human colon carcinoma LoVo/5-Fu cells. Expression of MDR1 mRNA was detected by real-time quantitative PCR. Drug sensitivity was measured by MTT. The apoptosis of cells was determined by flow cytometry assay. Results  After transfection with EASY-shRNA-MDR1,compared with the un-transfected group,expression of MDR1 mRNA in LoVo/5-Fu cells was reduced obviously(P<0.05); IC50 of 5-Fu and the resistance indexes were decreased obviously(P<0.05); the relative reverse rate of sensitivity of LoVo/5-Fu cells to 5-Fu was 74.7%; the apoptosis rate increased significantly (P<0.05). Conclusion  MDR1 shRNA effectively inhibited expression of MDR1,thus reuersed MDR1 genemediated multidrug resistance in human colon carcinoma LoVo/5-Fu cells.

Key words: Colon neoplasms; RNA interference; Multidrug resistance gene; Apoptosis; Gene therapy

中图分类号: 

  • R735.3
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