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RGD-TAT-KDRsiRNA慢病毒载体构建及抗肿瘤活性

张俭丽 郑旭 王杰

张俭丽, 郑旭, 王杰. RGD-TAT-KDRsiRNA慢病毒载体构建及抗肿瘤活性[J]. 中国公共卫生, 2015, 31(5): 594-597. doi: 10.11847/zgggws2015-31-05-15
引用本文: 张俭丽, 郑旭, 王杰. RGD-TAT-KDRsiRNA慢病毒载体构建及抗肿瘤活性[J]. 中国公共卫生, 2015, 31(5): 594-597. doi: 10.11847/zgggws2015-31-05-15
ZHANG Jian-li, ZHENG Xu, WANG Jie. Lentiviral vector construction of RGD- TAT-KDR siRNA fusion gene and assessment of its antitumor activity[J]. Chinese Journal of Public Health, 2015, 31(5): 594-597. doi: 10.11847/zgggws2015-31-05-15
Citation: ZHANG Jian-li, ZHENG Xu, WANG Jie. Lentiviral vector construction of RGD- TAT-KDR siRNA fusion gene and assessment of its antitumor activity[J]. Chinese Journal of Public Health, 2015, 31(5): 594-597. doi: 10.11847/zgggws2015-31-05-15

RGD-TAT-KDRsiRNA慢病毒载体构建及抗肿瘤活性

doi: 10.11847/zgggws2015-31-05-15
基金项目: 

辽宁省自然科学基金(201102214);沈阳市科技局科技计划项目(F11-264-1-29)

详细信息
    作者简介:

    张俭丽(1964),女,辽宁兴城人,高级讲师,本科,研究方向:抗肿瘤治疗。

    通信作者:

    王杰,E-mail:wangjie19932002@163.com

  • 中图分类号: R373.9

Lentiviral vector construction of RGD- TAT-KDR siRNA fusion gene and assessment of its antitumor activity

  • 摘要: 目的构建精氨酰-甘氨酰-天冬氨酸(RGD)介导的穿膜肽(TAT)-血管内皮细胞生长因子受体(KDR) siRNA融合基因慢病毒载体, 探讨其对人肺癌A549细胞的抑制效果。方法设计合成编码RGD的2条寡核苷酸链, 克隆到pGC/TAT-KDR siRNA载体的TAT下游, 构建TAT-RGD-KDR siRNA融合基因载体;慢病毒包装并感染A549细胞;通过Western blot和real-time PCR检测A549细胞KDR基因表达水平;通过噻唑蓝法、双色法流式细胞仪和Transwell侵袭实验检测KDR基因沉默对A549细胞凋亡和侵袭力的影响。结果测序和Blast比对证实重组载体中的RGD序列与设计一致;TAT-RGD-KDR siRNA融合基因慢病毒载体转染A540细胞KDR mRNA和蛋白表达水平分别为(22.7±3.9)%和(19.3±2.7)%, 明显低于TAT-KDR siRNA融合基因慢病毒载体转染组, 差异有统计学意义(P<0.01);该载体具有较强的抑制A549细胞增殖、促进细胞凋亡和抑制细胞体外侵袭力的作用。结论TAT-RGD-KDR siRNA融合基因慢病毒载体通过抑制细胞KDR mRNA和蛋白表达, 抑制A549细胞增殖和侵袭、促进细胞凋亡。
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    [9] Xiao B,Li W,Yang J,et al.RGD-IL-24,a novel tumor-targeted fusion cytokine:expression,purification and functional evaluation[J].Mol Biotechnol,2009,41(2):138-144.
    [10] Pesonen S,Diaconu I,Cerullo V,et al.Integrin targeted oncolytic adenoviruses Ad5-D24-RGD and Ad5-RGD-D24-GMCSF for treatment of patients with advanced chemotherapy refractory solid tumors[J].Int J Cancer,2012, 130(8):1937-1947.
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出版历程
  • 接收日期:  2014-07-01
  • 刊出日期:  2015-05-10

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