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miR‐186 suppresses cell proliferation and anchorage‐independence in a metastatic prostate cancer cell line
Author(s) -
Jones Dominique Z.,
Schmidt M. Lee,
Hobbing Katharine R.,
Clark Geoffrey,
Kidd LaCreis R.
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.936.6
Subject(s) - prostate cancer , cell growth , ectopic expression , microrna , cancer research , biology , cell culture , transfection , cancer , downregulation and upregulation , cell , microbiology and biotechnology , gene , genetics , biochemistry
MicroRNA (miR) dysregulation alters the expression of cancer related genes and contributes to disease states in many cancers. For example, ectopic miR‐186 expression leads to enhanced cell proliferation and migration in pancreatic cancer. However, the role of miR‐186 in prostate cancer (PCa) remains unclear. Previously, we observed significant upregulation of miR‐186‐5p in PCa patient serum (stage III/IV) relative to non‐cancerous controls. Furthermore, miR‐186 was significantly up‐regulated in metastatic PCa (PC‐3) cells compared to normal prostate epithelial cells (RWPE1). We hypothesized miR‐186 inhibition will reduce aggressive PCa in vitro . Consequently, miR‐186 was transiently and stably inhibited in PC‐3 cells. Cell proliferation and colony formation were evaluated for 7 and 21 days via Trypan Blue exclusion and soft agar assays. Aberrant gene expression was evaluated in transfected cells to identify miR‐186 targets. Candidate miR‐186 targets were selected using published reports on ‘miR‐186 and cancer’, availability of robust antibodies, and statistical filtering (false discovery ≥0.05 and ±1.2 fold change). Mir‐186 inhibition in PC‐3 cells significantly repressed proliferation and colony formation by 34–64%. Following miR‐186 inhibition in PC‐3 cells, 2,343 identified mRNA targets were differentially expressed compared to scramble controls (p < 0.05). The target list was reduced to 11 up‐regulated candidates. Predicted miR‐186 targets are undergoing validation via qRT‐PCR, western blots, and luciferase reporter assays. In addition, other studies are in progress to evaluate the impact of miR‐186 on cellular migration and invasion. Such efforts may lead to the identification of novel biomarkers to improve detection and clinical management strategies for aggressive PCa. Support or Funding Information Research & Fellowship Support: National Cancer Institute Grant R25‐CA‐134283 to DWH; Our Highest Potential Endowment in Cancer Research to LRK