miR-497-5p inhibits cell proliferation and invasion by targeting KCa3.1 in angiosarcoma

Oncotarget. 2016 Sep 6;7(36):58148-58161. doi: 10.18632/oncotarget.11252.

Abstract

Angiosarcoma is a rare malignant mesenchymal tumor with poor prognosis. We aimed to identify malignancy-associated miRNAs and their target genes, and explore biological functions of miRNA and its target in angiosarcoma. By miRNA microarrays and reverse transcription polymerase chain reaction, we identified 1 up-regulated miRNA (miR-222-3p) and 3 down-regulated miRNAs (miR-497-5p, miR-378-3p and miR-483-5p) in human angiosarcomas compared with human capillary hemangiomas. The intermediate-conductance calcium activated potassium channel KCa3.1 was one of the putative target genes of miR-497-5p, and marked up-regulation of KCa3.1 was detected in angiosarcoma biopsy specimens by immunohistochemistry. The inverse correlation of miR-497-5p and KCa3.1 also was observed in the ISO-HAS angiosarcoma cell line at the mRNA and protein levels. The direct targeting of KCa3.1 by miR-497-5p was evidenced by reduced luciferase activity due to complementary binding of miR-497-5p to KCa3.1 mRNA 3' untranslated region. For the functional role of miR-497-5p/KCa3.1 pair, we showed that application of TRAM-34, a specific KCa3.1 channel blocker, or transfection of ISO-HAS cells with KCa3.1 siRNA or miR-497-5p mimics inhibited cell proliferation, cell cycle progression, and invasion by down-regulating cell-cycle related proteins including cyclin D1, surviving and P53 and down-regulating matrix metallopeptidase 9. In an in vivo angiosarcoma xenograft model, TRAM-34 or miR-497-5p mimics both inhibited tumor growth. In conclusion, the tumor suppressor miR-497-5p down-regulates KCa3.1 expression and contributes to the inhibition of angiosarcoma malignancy development. The miR-497-5p or KCa3.1 might be potential new targets for angiosarcoma treatment.

Keywords: cell cycle; hemangioma; potassium channel; sarcoma.

MeSH terms

  • 3' Untranslated Regions
  • Animals
  • Carcinogenesis / genetics
  • Cell Cycle / drug effects
  • Cell Cycle Proteins / metabolism
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cell Proliferation / genetics
  • Down-Regulation
  • Female
  • Gene Expression Regulation, Neoplastic*
  • Genes, Tumor Suppressor*
  • Hemangiosarcoma / genetics*
  • Hemangiosarcoma / pathology
  • Humans
  • Immunohistochemistry
  • Intermediate-Conductance Calcium-Activated Potassium Channels / genetics*
  • Intermediate-Conductance Calcium-Activated Potassium Channels / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Microarray Analysis
  • Neoplasm Invasiveness / genetics
  • Pyrazoles / pharmacology
  • RNA Interference
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism
  • Transfection
  • Up-Regulation

Substances

  • 3' Untranslated Regions
  • Cell Cycle Proteins
  • Intermediate-Conductance Calcium-Activated Potassium Channels
  • KCNN4 protein, human
  • MIRN497 microRNA, human
  • MicroRNAs
  • Pyrazoles
  • RNA, Messenger
  • RNA, Small Interfering
  • TRAM 34