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

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.

• Authors: Yaobing Chen, Dong Kuang, Xia Zhao, Dong Chen, Xiaoyan Wang, Qin Yang, Jie Wan, Yuanli Zhu, Yu Wang, Shiying Zhang, Ying Wang, Qiang Tang, Mikio Masuzawa, Guoping Wang, Yaqi Duan
• Journal: Oncotarget (Oncotarget) Vol. 7 Issue 36 Pg. 58148-58161 (09 06 2016) ISSN: 1949-2553 [Electronic] United States
• PMID: 27531900 (Publication Type: Journal Article)

Chemical References

• 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

Topics

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