| A Calcium-sensing Receptor in Primary Cultured Human Osteoblast-like Cells?: Preliminary Study |
| 배양된 사람 조골세포에서 칼슘수용체에 관한 연구 |
| 윤현구,황창선*,강영순*,문인걸*,오한진**,임창훈,정호연,한기옥,장학철,안재용***,한인권 |
| 성균관의대 삼성제일병원 내과, 내분비연구실*, 가정의학과**, 정형외과*** |
|
|
| Abstract |
Objective
s: Calcium sensing receptor (CaR) has been found in different tissues such as parathyroid, kidney, C cell of thyroid, intestinal epithelium, bone marrow cells, osteoclast, and blood monocytes, and CaRs are known to respond to changes in extracellular calcium ions, calcium ions and CaR are very important for a variety of biologic functions in the body. But the presence of CaR in osteoblasts is still controversial and no study reported the presence of CaR in human osteoblasts. The purpose of this study is to evaluate whether CaRs are expressed or not in the cultured human osteoblasts.
Methods
Normal human bone cells were isolated from the bony fragments of human vertebrae. Western blot analysis with polyclonal anti-CaR antibody, reverse-transcriptase polymerase chain reaction (RT-PCR), Northern blot analysis were carried out to detect CaR in primary cultured human osteoblastlike cells. And RT-PCR products were sequenced.
Results
Multiple CaR immunoreactive bands were detected on Western blot analysis which is inconsistent with prior reports. Nucleotide sequencing of RT-PCR products corresponded to those of human kidney CaR complementary DNA (cDNA). On Northern blot analysis contained transcript of 5.2 kb similar to other tissues expressed CaR and additional 2.9 kb transcript.
Conclusion
Based on this preliminary data, we suggest the CaR might be exist in primary cultured human osteoblast-like cells although we could not confirm biochemically.
|
| Key Words:
Calcium sensing receptor (CaR), Human osteoblast-like cells, Western blot, RT-PCR, Direct sequencing, Northern blot |
|
PDF Links
Full text via DOI
Download Citation
