MicroRNA-141 regulates the tumour suppressor DLC1 in colorectal cancer.
MicroRNA-141 regulates the tumour suppressor DLC1 in colorectal cancer.
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DOI：
10.4149/neo_2015_084
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发表时间：
2015
期刊：
影响因子：
3
通讯作者：
Wu Pp;Zhu Hy;Sun Xf;Chen Lx;Q. Zhou;Jie Chen
中科院分区：
文献类型：
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作者：
Wu Pp;Zhu Hy;Sun Xf;Chen Lx;Q. Zhou;Jie Chen
我们以前的研究表明，DLC 1在结直肠癌（CRC）细胞系中作为一种功能性肿瘤抑制因子。本研究的目的是确定DLC 1是否是MicroRNA（miRNA）调控的靶点，并评估这种机制在CRC中的作用。通过生物信息学方法和文献分析，选择miR-141作为研究对象。合成miR-141模拟物、miR-141抑制剂并转染Lovo细胞。MTT法和体内模型法测定细胞生长情况。流式细胞仪检测细胞周期，transwell法检测细胞侵袭能力。荧光素酶报告基因分析和Western印迹显示DLC 1是miR-141在CRC中的直接靶点。miR-141在结直肠癌组织中的表达水平明显高于癌旁组织，而DLC 1在大部分临床标本中的表达水平下调（14/18）。此外，相关性分析显示CRC组织中miR-141水平与DLC 1表达水平之间呈负相关。MiR-141过表达促进Lovo细胞体内外生长，促进细胞周期进程和侵袭能力。此外，在miR-141过表达的Lovo细胞中重新引入DLC-1降低了细胞的生长速率，增加了G 0/G1期的百分比，并减少了迁移细胞的数量。总之，我们证明了miR-141在CRC中上调，并通过靶向DLC 1作为功能性癌基因。
Our previous study has showed that DLC1 acts as a functional tumor suppressor in colorectal cancer (CRC) cell lines. The aims of this study were to determine whether DLC1 is a target of MicroRNA (miRNA) regulation and to evaluate the role of this mechanism in CRC. By bioinformatics approach and literature, miR-141 was chosen for further study. The miR-141 mimic, miR-141 inhibitor were synthesized and transfected to Lovo cells. Cell growth was determined by MTT and in vivo models. The flow cytometric analysis for cell cycle determination and transwell assays for evaluating the cell invasion were used. Luciferase reporter assays and Western blots showed that DLC1 was a direct target of miR-141 in CRC. The expression levels of miR-141 were obviously up-regulated in CRC tissues compared to non-cancerous tissues, while DLC1 expression levels were down-regulated in a high proportion of clinical samples (14/18). In addition, correlation analyses revealed negative correlation between miR-141 levels and DLC1 expression levels in CRC tissues. MiR-141 overexpression promoted cell growth in vitro and in vivo, promoted cell cycle progression and invasion in Lovo cells. Furthermore, re-introduction of DLC-1 in miR-141-overexpressing Lovo cells decreased growth rate of cells, increase of the percentage in G0/G1 phase and decreased the number of migrating cells. In conclusion, we demonstrated that miR-141 is up-regulated in CRC and acts as a functional oncogene by targeting DLC1.