| P65基因对人三阴性乳腺癌细胞MDA-MB-231体外侵袭的影响 |
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| 引用本文： | 王玲,单保恩,桑梅香,连易水,丁春艳,周欣亮.P65基因对人三阴性乳腺癌细胞MDA-MB-231体外侵袭的影响[J].中国肿瘤生物治疗杂志,2011,18(5):502-507. |
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| 作者姓名： | 王玲 单保恩 桑梅香 连易水 丁春艳 周欣亮 |
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| 作者单位： | 河北医科大学第四医院肿瘤研究所免疫室,河北石家庄,050011 |
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| 基金项目： | 河北省科学技术研究与发展计划项目资助（No. 10276167） |
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| 摘 要： | 目的:观察miRNA靶向沉默P65基因对人三阴性乳腺癌(triple-negative breast cancer,TNBC)MDA-MB-231细胞体外黏附和侵袭的影响及其可能机制。方法:设计3对针对P65基因的特异性miRNA(P65miRNA-1、P65miRNA-2和P65miRNA-3),转染MDA-MB-231细胞,Western blotting检测P65蛋白的表达,筛选沉默效果最好的miRNA进行后续实验。体外黏附实验和Transwell小室实验检测P65miRNA转染前后MDA-MB-231细胞的黏附和侵袭。RT-PCR检测MDA-MB-231细胞基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)、MMP-9 mRNA的表达水平,明胶酶谱法检测MDA-MB-231细胞中MMP-2、MMP-9的酶活性。结果:成功构建重组质粒P65miRNA-1、P65miRNA-2和P65miRNA-3,前两者转染MDA-MB-231细胞后抑制P65蛋白表达80%以上。P65miRNA-1和P65miRNA-2转染对MDA-MB-231细胞的黏附无明显影响(P>0.05),但显著抑制MDA-MB-231细胞的侵袭(0.371±0.039、0.309±0.046 vs 0.698±0.065,P<0.05)。P65miRNA-1和P65miRNA-2转染沉默P65表达后,MDA-MB-231细胞MMP-2 mRNA(0.281±0.018、0.478±0.023 vs 1.056±0.072、1.128±0.059,P<0.05)和MMP-9 mRNA表达(0.193±0.013、0.371±0.035 vs 1.206±0.069、1.089±0.057,P<0.05)及其活性均显著下调。结论:miRNA靶向沉默P65的表达能抑制人TNBC MDA-MB-231细胞的体外侵袭,其机制可能与抑制MMP-2、MMP-9的表达和活性有关。
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| 关 键 词： | 三阴性乳腺癌 miRNA 侵袭 P65基因 MMP-2 MMP-9 |
| 收稿时间： | 2011/7/28 0:00:00 |
| 修稿时间： | 9/9/2011 12:00:00 AM |
P65 gene on invasion of human triple negative breast cancer cell line MDA MB 231 |
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| WANG Ling,SHAN Bao en,SANG Mei xiang,LIAN Yi shui,DING Chun yan and ZHOU Xin liang.P65 gene on invasion of human triple negative breast cancer cell line MDA MB 231[J].Chinese Journal of Cancer Biotherapy,2011,18(5):502-507. |
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| Authors: | WANG Ling SHAN Bao en SANG Mei xiang LIAN Yi shui DING Chun yan and ZHOU Xin liang |
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| Institution: | Department of Immunology, Tumor Research Institute, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China;Department of Immunology, Tumor Research Institute, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China;Department of Immunology, Tumor Research Institute, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China;Department of Immunology, Tumor Research Institute, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China;Department of Immunology, Tumor Research Institute, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China;Department of Immunology, Tumor Research Institute, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China |
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| Abstract: | Objective:To investigate the effect of targeting silence of P65 gene by miRNA on adhesion and invasion of human triple-negative breast cancer(TNBC) cell line MDA-MB-231 in vitro and its possible mechanism.Methods: Three pairs of miRNAs specifically targeting P65 gene(P65miRNA-1,P65miRNA-2 and P65miRNA-3) were designed and transfected into MDA-MB-231 cells,the level of P65 protein was detected by Western blotting,and P65miRNA with best silencing result was selected and used in the following experiments.The a... |
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| Keywords: | triple-negative breast cancer miRNA invasion P65 gene MMP-2 MMP-9 |
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