沉默RhoA基因对结肠癌LoVo细胞体内生长的抑制作用
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陈道瑾 E-mail:Chendj305@sohu.com

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湖南省科学技术厅科技计划资助(2008FJ3160)。


Inhibitory effect of RhoA gene silencing on growth of LoVo colon cancer cells in vivo
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    摘要:

    目的:RNA干扰使RhoA基因沉默后对结肠癌LoVo细胞在裸鼠体内生长的影响。 方法:构建靶向RhoA基因的shRNA真核表达载体,转染LoVo细胞,筛选RhoA基因沉默的稳定克隆。采用Western blot法检测RhoA蛋白的表达水平。将稳定转染RhoA shRNA表达载体、转染空载体和未转染的LoVo细胞分别接种于裸鼠皮下,4周后测量瘤体的体积和重量;免疫组化检测肿瘤组织RhoA,VEGF和MMP-2蛋白的表达。 结果:RhoA shRNA转染组瘤体的体积与瘤重明显低于其他两对照组(P<0.01)。免疫组化结果显示,RhoA shRNA染组瘤体组织RhoA,VEGF和MMP-2蛋白表达均较两对照组显著降低(P<0.01)。 结论:沉默RhoA可以抑制结肠癌LoVo细胞移植瘤的生长。

    Abstract:

    Objective:To investigate the effects of RhoA gene silencing by RNA interference on the growth of LoVo colon cancer cells transplanted into nude mice. Methods:The eukaryotic expression vectors of shRNA targeting RhoA were constructed and transfected into LoVo cells. The stable cell colons of RhoA silencing were selected, and the expression of RhoA protein was determined by Western blot. The nude nice were transplanted by subcutaneous inoculation with stably transfected RhoA shRNA plasmid, empty vector and untransfected LoVo cells, respectively. The weight and volumes of tumors were measured 4 weeks after transplantation. The expressions of RhoA, VEGF and MMP-2 in tumor tissue were examined by immunohistochemical staining. Results:The weight and volume of tumor in RhoA shRNA group was significantly lower than that of two control groups (P<0.01). The immunohistochemical results demonstrated that the expressions of RhoA, VEGF and MMP-2 protein were all significantly decreased compared with two control groups (P<0.01). Conclusions:RhoA gene silencing can inhibit the growth of LoVo cell xenograft tumnor in vivo.

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杨开焰| 陈道瑾| 李小荣| 文言广.沉默RhoA基因对结肠癌LoVo细胞体内生长的抑制作用[J].中国普通外科杂志,2011,20(4):383-386.
DOI:10.7659/j. issn.1005-6947.2011.04.017

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  • 收稿日期:2010-03-29
  • 最后修改日期:2010-10-09
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  • 在线发布日期: 2011-04-15