MicroRNA-338-3p inhibits colorectal carcinoma cell invasion and migration by targeting smoothened.

OBJECTIVE To investigate the regulative effect of microRNA-338-3p on colorectal carcinoma cell invasion and migration. METHODS The microRNA-338-3p expression pattern of colorectal carcinoma tissues and cell lines was detected by real-time reverse transcriptase polymerase chain reaction. The protein level of smoothened was detected by western blot analysis. Furthermore, colorectal carcinoma cells were pretreated with or without anti-smoothened-small interfering ribonucleic acid prior to the addition of pre-microRNA-338-3p or anti-microRNA-338-3p. The status of colorectal carcinoma cell invasion and that of migration were detected by transwell assay and wound healing assay, respectively. RESULTS The expression of microRNA-338-3p was significantly down-regulated in colorectal carcinoma tissues in comparison with those in the adjacent non-tumorous tissues, and the value was negatively related to advanced tumor, node, metastasis stage and local invasion. The expression of microRNA-338-3p in colorectal carcinoma cells transfected with pre-microRNA-338-3p p was significantly increased. Furthermore, over-expression of microRNA-338-3p inhibited the expression of smoothened protein in colorectal carcinoma cells, which showed obviously suppressed invasion and migration ability. The expression of microRNA-338-3p in colorectal carcinoma cells transfected with anti-microRNA-338-3p was significantly decreased. Moreover, the down-regulated expression of microRNA-338-3p caused the up-regulated expression of smoothened protein in colorectal carcinoma cells, which showed significantly enhanced invasion and migration ability. However, anti-smoothened-small interfering ribonucleic acid largely, but not completely, reversed the effects induced by blockage of microRNA-338-3p, suggesting that the regulative effect of microRNA-338-3p on colorectal carcinoma cell invasion and migration was indeed mediated by smoothened. Additionally, smoothened was identified as a direct target of microRNA-338-3p by luciferase assay. CONCLUSIONS MicroRNA-338-3p could inhibit colorectal carcinoma cell invasion and migration by inhibiting smoothened expression.

[1]  J. Ju.  miRNAs as biomarkers in colorectal cancer diagnosis and prognosis. , 2010, Bioanalysis.

[2]  K. Arita,et al.  Role of sonic hedgehog signaling in migration of cell lines established from CD133-positive malignant glioma cells , 2011, Journal of Neuro-Oncology.

[3]  M. Kerin,et al.  MicroRNA expression in colorectal cancer. , 2012, Cancer biomarkers : section A of Disease markers.

[4]  John Q. Davies,et al.  Leukocyte adhesion-GPCR EMR2 is aberrantly expressed in human breast carcinomas and is associated with patient survival. , 2011, Oncology reports.

[5]  Lentivirus-mediated SMO RNA interference inhibits SMO expression and cell proliferation, and affects the cell cycle in LNCaP and PC3 cancer cell lines. , 2010, Asian journal of andrology.

[6]  J. Houghton,et al.  Hedgehog signaling drives cellular survival in human colon carcinoma cells. , 2011, Cancer research.

[7]  M. Fabbri.  miRNAs as molecular biomarkers of cancer , 2010, Expert review of molecular diagnostics.

[8]  Domenico Coppola,et al.  MicroRNA-155 Is Regulated by the Transforming Growth Factor β/Smad Pathway and Contributes to Epithelial Cell Plasticity by Targeting RhoA , 2008, Molecular and Cellular Biology.

[9]  M. Odenthal,et al.  Altered levels of the onco-microRNA 21 and the tumor-supressor microRNAs 143 and 145 in advanced rectal cancer indicate successful neoadjuvant chemoradiotherapy. , 2011, International journal of oncology.

[10]  Qian Wang,et al.  Bead‐based microarray analysis of microRNA expression in hepatocellular carcinoma: miR‐338 is downregulated , 2009, Hepatology research : the official journal of the Japan Society of Hepatology.

[11]  Yan Luo,et al.  Computational prediction of amphioxus microRNA genes and their targets. , 2009, Gene.

[12]  G. Nuovo,et al.  MicroRNA-126 inhibits invasion in non-small cell lung carcinoma cell lines. , 2008, Biochemical and biophysical research communications.

[13]  Xiaoying Shen,et al.  MicroRNA expression profiles in human colorectal cancers with liver metastases. , 2011, Oncology reports.

[14]  K. Sun,et al.  Construction of lentivirus-based inhibitor of hsa-microRNA-338-3p with specific secondary structure , 2012, Acta Pharmacologica Sinica.

[15]  A. Schetter,et al.  The Role of MicroRNAs in Colorectal Cancer , 2012, Cancer journal.

[16]  X. Che,et al.  Sonic Hedgehog/GLI1 signaling pathway inhibition restricts cell migration and invasion in human gliomas , 2010, Neurological research.

[17]  Jing-Song Chen,et al.  miR‐338‐3p suppresses invasion of liver cancer cell by targeting smoothened , 2011, The Journal of pathology.

[18]  Sakari Knuutila,et al.  MicroRNA profiling differentiates colorectal cancer according to KRAS status , 2012, Genes, chromosomes & cancer.

[19]  G. Goodall,et al.  The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1 , 2008, Nature Cell Biology.

[20]  Gui-ying Zhang,et al.  Prognostic value of hedgehog signaling pathway in patients with colon cancer , 2012, Medical Oncology.

[21]  Paul Ahlquist,et al.  MicroRNA 29c is down-regulated in nasopharyngeal carcinomas, up-regulating mRNAs encoding extracellular matrix proteins , 2008, Proceedings of the National Academy of Sciences.

[22]  M. Neurath,et al.  Colitis and Colorectal Cancer , 2012, Digestive Diseases.

[23]  J. Houghton,et al.  Blocking Hedgehog survival signaling at the level of the GLI genes induces DNA damage and extensive cell death in human colon carcinoma cells. , 2011, Cancer research.

[24]  Hai-Jun Deng,et al.  miRNA-338-3p suppresses cell growth of human colorectal carcinoma by targeting smoothened. , 2013, World journal of gastroenterology.

[25]  A. Aschrafi,et al.  MicroRNA-338 Regulates Local Cytochrome c Oxidase IV mRNA Levels and Oxidative Phosphorylation in the Axons of Sympathetic Neurons , 2008, The Journal of Neuroscience.

[26]  Wei Wang,et al.  MicroRNA-221 inhibits CDKN1C/p57 expression in human colorectal carcinoma , 2011, Acta Pharmacologica Sinica.

[27]  A. Millar,et al.  Silencing the silencer: strategies to inhibit microRNA activity , 2011, Biotechnology Letters.