| Gantao Chen,Yanxiang Cheng,Xiao Yang,Liu Yaodan,Weiguo Dong. Targeting of RhoE inhibits epithelial-mesenchymal transition during colorectal cancer cell migration. Oncol Transl Med, 2016, 2: 119-126. |
| 靶向RhoE抑制上皮-间质转化对结直肠癌细胞迁移的影响及机制研究 |
| Targeting of RhoE inhibits epithelial-mesenchymal transition during colorectal cancer cell migration |
| Received:December 14, 2015 Revised:June 10, 2016 |
| DOI：10.1007/s10330-015-0124-6 |
| 中文关键词: miR-200b; 大肠癌; 金属蛋白酶; 上皮-间质化生; 细胞迁移 |
| 英文关键词: miR-200b; colorectal cancer (CRC); metalloproteinase (MMP); epithelial-mesenchymal transition (EMT); cell migration |
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| 中文摘要: |
| 摘要 目的：尽管微小RNA（miR-200b）已被证明可以促进结肠直肠癌（CRC）细胞的增殖，CRC细胞的miR-200b和上皮-间质化生（EMT）之间的关系仍然知之甚少。材料和方法：在大肠癌细胞（SW620和HT-29）中，miR-200b对EMT相关的标记物E钙粘蛋白和波形蛋白的影响，是由Western-blot印迹处理miR-200b类似物和抑制剂进行评价的。荧光素酶报告基因检测法用来检测下游的miR-200b目标。 Transwell小迁移测定法用来检测的CRC细胞迁移。结果：Western印迹显示，用miR-200b的类似物治疗导致E-钙粘蛋白上调和波形蛋白、金属蛋白酶（MMP）-9和MMP-2的下调，而用miR-200b的抑制剂治疗则表现出对E-cadherin和波形蛋白相反的效果。荧光素酶报告基因分析发现，RhoE（RND3）是miR-200B的靶点。在RhoE的3'UTR中，miR-200b的靶位点的预测目标站点1从 1584至1591，预测目标站点2从 1729至1735。 RhoE细胞系用来研究RhoE和miR 200b的对EMT和细胞迁移的影响。 RhoE增强了miR-200b的类似物上调E-钙粘蛋白和下调波形蛋白的效应。 RhoE也抑制了细胞迁移。此外，miR-200b的类似物治疗进一步促进RhoE对细胞迁移的抑制作用。结论：miR-200B通过在CRC抑制RhoE的表达来抑制EMT和CRC细胞迁移。 RhoE和miR-200b的可能是治疗大肠癌的靶基因。 |
| 英文摘要: |
| Objective: Despite microRNA (miR-200b) being proved to promote the proliferation of colorectal cancer
(CRC) cells, the relationship between miR-200b and epithelial-mesenchymal transition (EMT) of CRC cells
remains poorly understood. The aim of the study was to investigate the relationship between miR-200b and
EMT during CRC cell migration.
Methods: The effect of miR-200b on EMT-associated markers E-cadherin and vimentin was evaluated by
western blot in CRC cells (SW620 and HT-29) by treatment with miR-200b mimics and inhibitors. A luciferase
reporter assay was employed to detect downstream targets of miR-200b. Transwell migration assays
were used to detect CRC cell migration.
Results: Western blots revealed that treatment with miR-200b mimics led to up-regulation of E-cadherin
and down-regulation of vimentin, metalloproteinase (MMP)-9, and MMP-2, whereas treatment with miR-
200b inhibitor exhibited opposite effects on expression of E-cadherin and vimentin. Luciferase reporter
assays demonstrated that RhoE (RND3) was targeted by miR-200b. Two predicted target sites of miR-200b
were present in the 3’-UTR of RhoE. Predicted target site 1 was from nucleotides 1584 to 1591, and site
2 was from nucleotides 1729 to 1735. RhoE knockdown cell lines were also established to investigate the
impact of RhoE and miR-200b on EMT and cell migration. RhoE knockdown enhanced the effect of miR-
200b mimics, up-regulating E-cadherin and down-regulating vimentin. RhoE knockdown also inhibited cell
migration. Furthermore, miR-200b mimic treatment further promoted the inhibitory effect of RhoE knockdown
on cell migration.
Conclusion: miR-200b inhibited EMT and CRC cell migration partly via inhibiting RhoE expression in
CRC. RhoE and miR-200b might therefore be promising target genes in the management of CRC. |
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