lncRNA HOTAIR Inhibits Mineralization in Osteoblastic Osteosarcoma Cells by Epigenetically Repressing ALPL

Calcified Tissue International
Aya Misawa, Hideo Orimo

Abstract

HOTAIR is a lncRNA that plays critical role in gene regulation and chromatin dynamics through epigenetic mechanisms. In this work we studied the physiological role of HOTAIR during the process of mineralization using osteoblastic osteosarcoma cells focusing in ALPL (Tissue Non-Specific Alkaline Phosphatase), a pivotal gene that controls bone formation. HOTAIR knockdown resulted in upregulation of ALPL, increase of alkaline phosphatase (ALP) activity, and enhanced mineralization in osteoblastic SaOS-2 cells cultured in mineralizing medium. Luciferase assays using reporter vectors containing ALPL promoter showed that HOTAIR repression increases ALPL promoter activity. Furthermore, HOTAIR knockdown increased histone H3K4 methylation levels at ALPL promoter region, suggesting that ALPL repression by HOTAIR is regulated by epigenetic mechanisms. This work supports that physiological bone formation is epigenetically regulated by a lncRNA.

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Citations

Jan 20, 2021·BMC Genomics·Jun FangHuan Min Zhou
May 28, 2021·BMC Bioinformatics·Swapna Vidhur DaulatabadSarath Chandra Janga

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Methods Mentioned

BETA
FCS
transfection
Assay
PCR
ChIP
histone acetylation

Software Mentioned

Integrative Genome Viewer ( IGV
FANTOM
siDirect

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