miR-124在结肠癌中表达及其对结肠癌肺转移的调控作用
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孙健勇, Email: sunjianyong2@163.com

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    国家自然科学基金资助项目(81802808);陕西省自然科学基础研究计划基金资助项目(2018JQ8050);陕西省教育厅科学研究计划项目(18JK1019)。


Expression of miR-124 in colon cancer and its regulatory effect on colon cancer lung metastasis
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    摘要:

    目的:探讨miR-124在结肠癌中的表达,及其对结肠癌细胞生物学行为的影响及作用机制。 方法:用qRT-PCR检测miR-124在结肠癌组织及细胞中的表达情况,用慢病毒载体转染技术上调miR-124在结肠癌细胞中的表达水平后,通过流式细胞术观察细胞凋亡的变化,用不同miR-124表达水平的结肠癌细胞对裸鼠尾静脉注射后观察肺转移及生存情况;通过生物信息学软件预测miR-124的潜在靶基因,并通过双荧光素酶报告基因分析以及体外、体内实验进行鉴定和验证。 结果:miR-124在结肠癌组织中表达降低,其表达水平与肿瘤大小、分期及淋巴结转移明显有关(均P<0.05)。miR-124表达水平随着结肠癌细胞恶性度的增加而降低,上调miR-124的表达后,结肠癌细胞凋亡率明显增加,裸鼠的肺转移明显抑制、生存时间明显延长(均P<0.05)。ITGB1是miR-124的潜在靶基因,下调结肠癌细胞ITGB1表达后,其作用与上调miR-124表达相一致。结肠癌组织中,miR-124与ITGB1的表达呈明显负相关(r=-0.4936,P=0.002)。 结论:在结肠癌中miR-124表达下调,其靶基因ITGB1表达相应增加,从而促进结肠癌细胞的生长和侵袭转移。

    Abstract:

    Objective: To investigate the expression of miR-124 in colon cancer and its actions on the biological behaviors of colon cancer cells as well as the potential mechanism. Methods: The expressions of miR-124 in colon cancer tissue and cells were examined by qRT-PCR. In colon cancer cells after up-regulating miR-124 expression through lentiviral vector transfection technique, the change in apoptosis was examined by flow cytometry. Nude mice were injected with colon cancer cells of different miR-124 expression levels, and then, the pulmonary metastasis and survival time were observed. The target genes of miR-124 were predicted by bioinformatic analysis, followed by identification and verification through dual luciferase reporter assay and a series of in vitro and in vivo experiments. Results: The miR-124 expression was significantly reduced in colon cancer tissue, and its expression was significantly associated with the tumor size, tumor stage and lymph node metastasis (all P<0.05). The miR-124 expression level was decreased with the increase of the degree of malignancy of the colon cancer cells. After up-regulation of miR-124 expression, the apoptosis of colon cancer cells was significantly increased, pulmonary metastasis was inhibited and survival time was prolonged in the nude mice (all P<0.05). ITGB1 was suggested as the potential target gene for miR-124. In colon cancer cells, down-regulating ITGB1 expression showed the similar effects as those exerted by up-regulating miR-124 expression. There was a negative correlation between miR-124 and ITGB1 expressions in colon cancer tissue (r=–0.4936, P=0.002). Conclusion: The miR-124 expression is down-regulated in colon cancer, with accordingly increased expression of its target gene ITGB1, which promotes the growth as well as the invasion and metastasis of the colon cancer cells.

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楚琰, 李沛, 杨光, 赵换维, 王雅台, 戴建国, 朱介法, 孙健勇. miR-124在结肠癌中表达及其对结肠癌肺转移的调控作用[J].中国普通外科杂志,2018,27(10):1279-1287.
DOI:10.7659/j. issn.1005-6947.2018.10.010

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  • 收稿日期:2018-05-25
  • 最后修改日期:2018-09-16
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  • 在线发布日期: 2018-10-25