microRNA-133b抑制人结直肠癌细胞迁移和侵袭的研究
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胡桂 E-mail:hugui22@qq.com

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教育部高等学校博士点科研基金资助项目(200805330040)。


The suppressive effect of microRNA-133b on migration and invasion of human colorectal carcinoma cells
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    摘要:

    目的:研究microRNA-133b在结直肠癌细胞中的表达及其对细胞迁移和侵袭能力的影响。
    方法:采用实时定量PCR检测microRNA-133b(miR-133b)在不同恶性程度的结直肠癌细胞及正常结肠细胞中的表达差异;构建miR-133b的真核表达载体并稳定转染SW-620细胞,筛选出稳定表达miR-133b及对照质粒的细胞株SW-620-133b和SW-620-HK,实时定量PCR检测转染后miR-133b表达的变化;划痕实验检测miR-133b对SW-620细胞迁移能力的影响;Transwell侵袭实验检测miR-133b对SW-620细胞侵袭能力的影响。
    结果:正常结肠细胞系CCD-18Co中miR-133b的表达显著高于4种结直肠癌细胞(SW-480,HT-29,LoVo,SW-620)(P<0.05),且其在结直肠癌细胞中的表达随着肿瘤细胞恶性程度的增加而降低。稳定转染后,SW-620-133b细胞miR-133b表达量较SW-620及SW-620-HK组显著增高(P<0.05);miR-133b对SW-620细胞迁移和侵袭能力有显著的抑制作用,抑制率分别为(42.2±2.6)%及(66.3±8.7)%。
    结论:miR-133b在结直肠癌细胞中呈低表达,miR-133b高表达能显著抑制SW-620细胞的迁移和侵袭能力。

    Abstract:

    Objective:To explore the expression of microRNA-133b in colorectal carcinoma cells and its effect on migration and invasion of colorectal cancer cells.
    Methods:The expression of microRNA-133b (miR-133b) in normal human colon cells and four colorectal cancer cell lines with different malignant potential were examined by real-time PCR. The miR-133b genomic sequences were amplified from human genomic DNA by PCR and constructed into the eukaryotic expression vector. SW-620 cells were transfected with the identified plasmid by Effectene. SW-620 cell lines with stable expression of miR-133b or control were obtained after a 3-week selection period in medium supplemented with G418, designated SW-620-133b and SW-620-HK, respectively. Cell migration and invasion was measured in vitro by wound healing assay and transwell assay.
    Results:miR-133b expression was shown to be greatly downregulated in human colon cancer cells (SW-480,HT-29,LoVo,SW-620) compared to normal colon cells CCD-18Co (P<0.05), and this downregulation was inversely associated with degree of malignancy of colorectal cancer cells. In cancer SW-620 cells, expression of miR-133b significantly inhibited tumor cell migration and invasion, with the inhibiting rates of (42.2±2.6) % and (66.3±8.7) %, respectively.
    Conclusions:The expression of miR-133b is downregulated in human colorectal cancer cells. Overexpression of miR-133b significantly inhibits SW-620 cell migration and invasion.

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胡桂|陈道瑾|李小荣|杨开焰|成伟. microRNA-133b抑制人结直肠癌细胞迁移和侵袭的研究[J].中国普通外科杂志,2010,19(4):359-363.
DOI:10.7659/j. issn.1005-6947.2010.04.007

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  • 收稿日期:2009-11-23
  • 最后修改日期:2010-01-21
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  • 在线发布日期: 2010-04-15