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miR-30b对结直肠癌细胞转移潜能的影响
引用本文:吴萍,冶亚平,丁彦青,廖雯婷.miR-30b对结直肠癌细胞转移潜能的影响[J].中国肿瘤临床,2014,41(11):679-683.
作者姓名:吴萍  冶亚平  丁彦青  廖雯婷
作者单位:南方医科大学基础医学院病理学系(广州市510515)
基金项目:国家自然科学基金项目81172055国家自然科学基金项目81071735国家自然科学基金项目U1201226
摘    要:   目的   明确miR-30b对结直肠癌细胞侵袭和迁移潜能的影响。   方法   RT-qPCR检测20对配对结直肠癌组织标本中miR-30b的表达;Transwell和划痕实验检测过表达和抑制miR-30b后结直肠癌细胞侵袭和迁移潜能的改变;应用生物信息学方法预测miR-30b的作用靶点;Western Blot及双荧光素酶报告基因实验验证过表达和抑制miR-30b后靶点Snail和EMT(epithelial mesenchymal transition)标志物的表达情况。   结果   癌组织中miR-30b表达水平明显低于正常肠黏膜组织;Transwell及划痕实验结果显示过表达miR-30b后结直肠癌细胞的侵袭迁移能力降低,抑制miR-30b后侵袭迁移能力增强;生物信息学预测结果显示miR-30b能够作用于Snail 3'-UTR,双荧光素酶检测结果进一步证实miR-30b能够作用于Snail 3'-UTR;Western Blot结果显示过表达miR-30b后Snail的表达下降,EMT标志物Vimentin表达下降和E-cadherin表达升高,而抑制miR-30b后结果相反。   结论   miR-30b在结直肠癌中表达水平下降,并通过靶向Snail调节结直肠癌细胞的侵袭和迁移。 

关 键 词:结直肠癌    miR-30b    侵袭    迁移    Snail
收稿时间:2014-04-23

The function of miR-30b in colorectal cancer metastasis
Institution:Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China
Abstract:   Objective   To determine the function of miR-30b in the metastasis of colorectal cancer cells.   Methods   RT-qPCR was performed to test miR-30b expression in 20 fresh primary colorectal cancer tissues and their corresponding adjacent tissues. Transwell and wound healing assays were performed to test the invasion and migration of colorectal cancer cells after miR-30b overexpression or inhibition. Bioinformatics assay was performed to predict miR-30b targets. Western Blot and Dual Luciferase reporter assay were performed to test the expressions of Snail and downstream target genes in colorectal cancer cells.   Results   The results reveal that miR-30b expression decreased in cancer tissues compared with normal tissues. Transwell and wound healing assays reveal that miR-30b overexpression inhibited cell invasion and migration, whereas miR-30b inhibition promoted the invasion and migration of colorectal cancer cells. Bioinformatics analyses reveal that miR-30b targets the 3'-UTR of Snail. Dual Luciferase reporter assay confirms that miR-30b affects the 3'-UTR of Snail. Western Blot analyses show that Snail and Vimentin expressions were significantly downregulated, whereas E-cadherin expression obviously increased after miR-30b overexpression. However, Snail and Vimentin expressions increased, but E-cadherin expression decreased after miR-30b inhibition.   Conclusions   Conclusion: The miR-30b gene is downregulated in colorectal cancer tissues. The miR-30b protein may be important in the regulation of cell invasion and migration by targeting Snail in colorectal cancer cells. 
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