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中华结直肠疾病电子杂志 ›› 2017, Vol. 06 ›› Issue (05) : 381 -386. doi: 10.3877/cma.j.issn.2095-3224.2017.05.006

所属专题: 文献

论著

在维吾尔族人群中探究miR-10b对结肠癌转移的作用与机制
苏莎莎1, 赫晓磊1, 高峰1,()   
  1. 1. 830000 新疆维吾尔自治区人民医院消化科
  • 收稿日期:2017-04-27 出版日期:2017-10-25
  • 通信作者: 高峰

Functions and regulation of miR-10b in Uyger colorectal cancer metastasis

Shasha Su1, Xiaolei He1, Feng Gao1,()   

  1. 1. Department of Gastroenterology, Xinjiang Uyger Muncipal People′Hospital, Urumchi Xinjiang 830000, China
  • Received:2017-04-27 Published:2017-10-25
  • Corresponding author: Feng Gao
  • About author:
    Corresponding author: Gao Feng, Email:
引用本文:

苏莎莎, 赫晓磊, 高峰. 在维吾尔族人群中探究miR-10b对结肠癌转移的作用与机制[J/OL]. 中华结直肠疾病电子杂志, 2017, 06(05): 381-386.

Shasha Su, Xiaolei He, Feng Gao. Functions and regulation of miR-10b in Uyger colorectal cancer metastasis[J/OL]. Chinese Journal of Colorectal Diseases(Electronic Edition), 2017, 06(05): 381-386.

目的

探究在多种肿瘤中均发挥重要调节作用的miR-10b对维吾尔族结肠癌细胞转移的影响。

方法

采用miR-10b过表达或抑制的结肠癌细胞系SW620,在细胞水平验证miR-10b对细胞转移能力的影响及对KLF4的调控作用。在临床样本组织中检测miR-10b的表达以佐证miR-10b功能。

结果

维吾尔族结肠癌组织按照确诊时有无淋巴结或远处转移分为转移组(14例)和未转移组(16例),结肠癌转移组miR-10b的表达较强(t=-5.372,P<0.05)。在SW620细胞中转染miR-10b抑制剂以敲低miR-10b,与对照细胞相比,miR-10b表达水平的降低使SW620细胞的侵袭以及迁移能力明显降低(t=26.56,10.40,P均<0.05)。在SW620细胞中转染miR-10b抑制剂以下调其表达,通过实时定量RT-PCR及Western Blot检测细胞中KLF4 mRNA水平和蛋白水平的改变,结果显示miR-10b表达水平的降低,细胞内KLF4 mRNA水平和蛋白水平升高(t=3.78,P<0.05),提示miR-10b对KLF4 mRNA的调节是从转录和翻译两个水平发生作用的。我们敲降miR-10b的表达或者增加KLF4的表达,通过检测E-cadherin、cyclins D1以及p53的改变,发现miR-10b可能通过影响涉及EMT、细胞周期和细胞凋亡的某些独立的信号通路,对结肠癌的细胞功能发挥重要作用。

结论

在维吾尔族人群中miR-10b能够促进结肠癌细胞SW620的增殖与迁移,参与结肠癌的侵袭与转移。

Objective

To investigate the role of miR-10b in metastasis of colon cancer of Uyger patients.

Methods

miR-10b expression was tested on CRC samples which were divided into metastatic groups and non-metastic groups of Uyger petients.The miR-10b-overexpressed or down-regulated SW620 cells were used to test the effects of miR-10b on invasion and migration of SW620 cells and to observe how miR-10b regulating KLF4. The miR-10b/klf4-overexpressed or down-regulated SW620 cells were used to test E-cadherin, cyclins D1 and p53 expression to test the involved mechanism in the regulatory function of miR-10b on metastasis and proliferation in CRC cells.

Results

miR-10b was over-expressed in metastatic CRC samples, compared with non-metastic ones of Uyger patients (t=-5.372, P<0.005). Inhibition of miR-10b in SW620 cells led to a notable reduction in invasion and migration assay compared with control cells (t=26.56, 10.40, P<0.05). We transfected miR-10b inhibitor into SW620 cells, an in crease of KLF4 mRNA level and protein level in SW620 cells was observed (t=3.78, P<0.05). miR-10b knockdown and KLF4 overexpression upregulated E-cadherin expression. Correlated with the modulated expression of miR-10b: the inhibition of miR-10b caused downegulation of cyclins D1 and p53, which were partly abrogated after co-transfecting with miR-10b and siRNA KLF4.

Conclusions

Expression of miR-10b is up-regulated in the metastasis CRC tissues and cells. miR-10b controls the metastasis and proliferation of CRC cells. miR-10b may exert its effect on metastasis and prolieration of CRC cells by regulating the expression of KLF4.miR-10b appears to modulate several independent signaling pathways that involved EMT, cell cycle and apoptosis.

表1 荧光定量PCR检测miR-10b在大肠癌组织中的表达情况
图1 荧光定量PCR检测miR-10b在大肠癌组织中的表达情况
表2 Transwell小室侵袭和迁移实验穿膜细胞计数
图2 倒置显微镜下观察SW620细胞在Transwell小室侵袭和迁移的情况
图3A 实时定量RT-PCR检测KLF4 mRNA水平的变化3B Western Blot检测KLF4蛋白水平的变化
图4 miR-10b对大肠癌的细胞功能发挥作用可能存在的机制
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