Induction of single nucleotide polymorphisms in the purinergic P2X7 receptor and subsequent investigation of changes in function and gene expression of specific bone markers

Nasir Ali & Barakat Ali

Studenteropgave: Speciale


Abstract Bone is a remarkable organ in the way of regulating and adapting to different mechanical loading. Any imbalance in the process of regulating and adapting, results in bone diseases such as osteoporosis. Since the discovery of P2 receptors in 1970s, research in the field has provided tremendous amount of information about the physiological and pathophysiological roles of P2 receptors in many human diseases. P2 receptors are divided in P2X and P2Y receptors whereas seven P2X subunits have been identified (P2X1-7). P2X7R gene is highly polymorphic and plays a role in the regulation of osteoblast activity and changes in receptor function may affect the osteoblast function in general. Several P2X7R SNPs have been associated with increased/decreased risk of fracture and lower/increased BMD in cohort studies dependent on whether it is a gain of function mutation or loss of function mutation. Therefore we decided to elucidate the role of various mutations in osteoblast function by inducing mutations in P2X7R gene and transfect the osteoblastic-like cell line ROS 17/2.8. The expression of P2X7R was determined in transfected cells and functional analysis was conducted to investigate the cellular effect of two gain of function (GOF) SNPs, H155Y and A348T and a single loss of function (LOF) R307Q. Furthermore the effect of these SNPs on gene expression of Runx2 and Sp7 and specific bone markers was also investigated. Expression of P2X7 mRNA was confirmed by Real-time qPCR and functional analysis revealed that A348T increased the receptor function while R307Q decreased the receptor function in accordance to exist data. The H155Y SNP showed a minor GOF although not significant. The gene expression of Runx2 and Sp7 did not show unambiguous results and the same applies for the bone markers. Our study is the first in terms of showing that R307Q and A348T SNP in P2X7R in osteoblastic cell line have LOF and GOF properties, respectively.

UddannelserMolekylærbiologi, (Bachelor/kandidatuddannelse) KandidatMedicinalbiologi, (Bachelor/kandidatuddannelse) Kandidat
Udgivelsesdato1 apr. 2014
VejledereSusanne Syberg & Ole Vang


  • Osteoporosis
  • P2X7 receptor