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ORIGINAL ARTICLE
Year : 2018  |  Volume : 14  |  Issue : 4  |  Page : 838-843

Post-transcriptional regulation DPC4 gene by miR-190 in colorectal cancer cells


1 Department of Pathology, Xiangya Hospital/School of Basic Medicine, Central South University, Hunan, China
2 Department of Pathology, The Second Xiangya Hospital, Central South University, Hunan, China
3 Cancer Research Institute, School of Basic Medicine, Center for Medicine Research, Xiangya Hospital, Central South University, Hunan, China

Correspondence Address:
Desheng Xiao
Department of Pathology, Xiangya Hospital, School of Basic Medicine, Central South University, Changsha 410078, Hunan
China
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jcrt.JCRT_577_17

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Objective: The objective of this study is to elucidate the regulation of the DPC4 gene by miR-190 in colorectal cancer (CRC) cells. The present study was undertaken to determine whether the DPC4 gene is a target gene of miRNA-190, identify target motifs and to elucidate the mechanism of regulation of DPC4 by miRNA-190. Materials and Methods: MiR-190 and DPC4 expression were measured in five different CRC cell lines by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. The regulation of DPC4 by miR-190 was evaluated by qRT-PCR, Western blotting, and luciferase reporter assays in the human CRC cell line HT-29 after treatment with miR-190 mimics and inhibitors. Results: The DPC4 mRNA, miR-, and DPC4 protein expression levels were highest in LS174T cells while lowest in SW480 and SW620 cells. The DPC4/miR-190 ratio in the HT-29 cancer cell line was the largest. MiR-190 expression increased dramatically after treatment with miR-190 mimics and decreased significantly after treatment with miR-190 inhibitors. DPC4 protein expression decreased in the miR-190 mimics transfection group when compared to the negative control (N.C.) group and increased in the miR-190 inhibitor groups when compared to the inhibitor plus N.C. group. MiR-190 inhibits the relative luciferase activity of psiCHECK-2™ vector-3'UTR compared to the N.C. group, while miR-190 had no obvious effect on the relative luciferase activity of the psiCHECK-2™ vector-3'UTRmut and psiCHECK-2™ vector transfected cells. Conclusions: The DPC4 gene might be the target gene of miR-190, which may negatively regulate the DPC4 gene in human CRC cells by translational suppression rather than mRNA degradation.


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