hsa-mir-30c promotes the invasive phenotype of metastatic breast cancer cells by targeting NOV/CCN3

Jason R Dobson; Hanna Taipaleenmäki; Yu-Jie Hu; Deli Hong; Andre J van Wijnen; Janet L Stein; Gary S Stein; Jane B Lian; Jitesh Pratap

doi:10.1186/s12935-014-0073-0

hsa-mir-30c promotes the invasive phenotype of metastatic breast cancer cells by targeting NOV/CCN3

Cancer Cell Int. 2014 Aug 2:14:73. doi: 10.1186/s12935-014-0073-0. eCollection 2014.

Authors

Jason R Dobson¹, Hanna Taipaleenmäki², Yu-Jie Hu³, Deli Hong⁴, Andre J van Wijnen⁵, Janet L Stein⁴, Gary S Stein⁴, Jane B Lian⁴, Jitesh Pratap⁶

Affiliations

¹ Department of Cell and Developmental Biology, University of Massachusetts Medical School, 55 Lake Ave, North, Worcester 01655, MA, USA ; Current address: Center for Computational Molecular Biology, Department of Molecular Biology, Cell Biology, and Biochemistry, and Department of Computer Science, Brown University, 115 Waterman Street, Providence 02912, RI, USA.
² Department of Cell and Developmental Biology, University of Massachusetts Medical School, 55 Lake Ave, North, Worcester 01655, MA, USA ; Current address: Heisenberg-Group for Molecular Skeletal Biology, Department of Trauma, Hand, and Reconstructive Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
³ Department of Cell and Developmental Biology, University of Massachusetts Medical School, 55 Lake Ave, North, Worcester 01655, MA, USA.
⁴ Department of Cell and Developmental Biology, University of Massachusetts Medical School, 55 Lake Ave, North, Worcester 01655, MA, USA ; Current address: Department of Biochemistry and Vermont Cancer Center, University of Vermont College of Medicine, 89 Beaumont Avenue, Burlington 05405-0068, VT, USA.
⁵ Department of Cell and Developmental Biology, University of Massachusetts Medical School, 55 Lake Ave, North, Worcester 01655, MA, USA ; Current address: Departments of Orthopedic Surgery and Biochemistry & Molecular Biology, Mayo Clinic, 200 First Street SW, Medical Sciences Building 3-69, Rochester 55905, MN, USA.
⁶ Department of Cell and Developmental Biology, University of Massachusetts Medical School, 55 Lake Ave, North, Worcester 01655, MA, USA ; Current address: Department of Anatomy and Cell Biology, Rush University Medical Center, Armour Academic Center, 600 S, Paulina Street, Suite 507, Chicago 60612, IL, USA.

Abstract

Background: For treatment and prevention of metastatic disease, one of the premier challenges is the identification of pathways and proteins to target for clinical intervention. Micro RNAs (miRNAs) are short, non-coding RNAs, which regulate cellular activities by either mRNA degradation or translational inhibition. Our studies focused on the invasive properties of hsa-mir30c based on its high expression in MDA-MB-231 metastatic cells and our bioinformatic analysis of the Cancer Genome Atlas that identified aberrant hsa-mir-30c to be associated with poor survival.

Methods: Contributions of hsa-mir-30c to breast cancer cell invasion were examined by Matrigel invasion transwell assays following modulation of hsa-mir-30c or hsa-mir-30c* levels in MDA-MB-231 cells. hsa-mir-30c in silico predicted targets linked to cell invasion were screened for targeting by hsa-mir-30c in metastatic breast cancer cells by RT-qPCR. The contribution to invasion by a target of hsa-mir-30c, Nephroblastoma overexpressed (NOV), was characterized by siRNA and invasion assays. Significant effects were determined using Student's T-tests with Welch's correction for unequal variance.

Results: MCF-7 and MDA-MB-231 cells were used as models of poorly invasive and late-stage metastatic disease, respectively. By modulating the levels of hsa-mir-30c in these cells, we observed concomitant changes in breast cancer cell invasiveness. From predicted targets of hsa-mir-30c that were related to cellular migration and invasion, NOV/CCN3 was identified as a novel target of hsa-mir-30c. Depleting NOV by siRNA caused a significant increase in the invasiveness of MDA-MB-231 cells is a regulatory protein associated with the extracellular matrix.

Conclusions: NOV/CCN3 expression, which protects cells from invasion, is known in patient tumors to inversely correlate with advanced breast cancer and metastasis. This study has identified a novel target of hsa-mir-30c, NOV, which is an inhibitor of the invasiveness of metastatic breast cancer cells. Thus, hsa-mir-30c-mediated inhibition of NOV levels promotes the invasive phenotype of MDA-MB-231 cells and significantly, the miR-30/NOV pathways is independent of RUNX2, a known target of hsa-mir-30c that promotes osteolytic disease in metastatic breast cancer cells. Our findings allow for mechanistic insight into the clinical observation of poor survival of patients with elevated hsa-mir-30c levels, which can be considered for miRNA-based translational studies.

Keywords: Hsa-mir-30c breast cancer; Invasion; Metastasis; NOV/CCN3; miRNA.

Abstract

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