Molecular Biology Reports

, Volume 46, Issue 2, pp 1667–1674 | Cite as

Epigenetic regulation of BMP2 gene in osteoporosis: a DNA methylation study

  • Mehrunnisa M. Raje
  • Richa AshmaEmail author
Original Article


Osteoporosis is a multifactorial disease in which genetic factors and epigenetic modifications play a major role. DNA methylation is known for gene silencing and its effect on BMP2 promoter has been studied here to understand its regulatory activity in osteoporosis pathogenicity. CpG methylation in the BMP2 promoter was analyzed by performing bisulfite specific PCR on the gDNA samples extracted from whole blood of osteoporotic (n = 24) and healthy (n = 24) individuals. Disproportionate allele frequency of CpG sites was calculated statistically. Differential BMP2 expression was estimated using quantitative RT-PCR technique. Luciferase reporter assay was performed to determine and confirm differential transcriptional activity of BMP2 promoter due to methylation. Total of 14 CpG sites were reporter in the BMP2 promoter of which, CpG site at − 267th position upstream to TSS was found to have disproportionate allele frequency among osteoporotic and healthy individuals and was found to be significantly associated with osteoporosis condition. Functional and gene expression analysis of this methylated site using luciferase reporter vector and Real Time PCR approach, suggested reduced transcriptional activity of BMP2 promoter as well as decreased gene expression in disease condition. BMP2 is being a central signaling molecule, aberrant methylation in the promoter region may result into down regulation of osteoblast markers involved in bone formation.


Osteoporosis Epigenetics BMP2 DNA methylation Luciferase reporter assay 



Alkaline phosphatase


Bone mineral density


Bone morphogenetic protein 2


BMP2 inducible kinase


Bone morphogenetic protein 4


Bone morphogenetic protein 6


Bone morphogenetic protein 7


Complementary DNA


Class interval




Distal-less homeobox 5


Deoxyribonucleic acid


Dual X-Ray absorptiometry


European Collection Of Authenticated Cell Cultures


Ethylenediaminetetraacetic acid


Eagle’s Minimum Essential Medium


Epithelial membrane protein 2


Extracellular signal-regulated kinases


Fold change


Glyceraldehyde 3-phosphate dehydrogenase


Genomic DNA


Indian Council of Medical Research




Luria agar


Luria Broth


Linkage disequilibrium


Messenger RNA


Multiple sequence alignment


National Center for Biotechnology Information




Peripheral blood mononuclear cell


Phosphate-buffered saline


Polymerase chain reaction


Quantitative real time polymerase chain reaction


Reference sequence database


Relative luciferase activity


Ribonucleic acid


Risk ratio


Reverse transcriptase polymerase chain reaction


Runt related transcription factor 2


Standard deviation


Single nucleotide polymorphisms


Activated kinase 1


Transcription factor


Transcription start site


World Health Organization


Compliance with ethical standards

Conflict of interest

The authors have declared that no conflict of interest exists.


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Center for Advanced Studies, Department of ZoologySavitribai Phule Pune UniversityPuneIndia

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