Obstetrics & gynecology science

Korean Society of Obstetrics and Gynecology (대한산부인과학회)
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Alendronate enhances osteoblastic differentiation with increased expression of Id-1 and Id-2 in pre-osteoblast cell line, MC3T3-E1

Alendronate에 의한 MC3T3-E1 골모세포주의 분화와 Id-1과 Id-2의 발현 증가

Park, Min-Jung;Joo, Bo-Sun;Lee, Ji-Eun;Kim, Hyun-Jung;Moon, Sung-Eun;Moon, Hwa-Sook
박민정;주보선;이지은;김현정;문성은;문화숙

  • Published : 20100300
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Abstract

Objective: Alendronate, a widely used bisphosphonates, acts to inhibit bone resorption by interfering with the activity of osteoclasts. Recently, it has been reported that alendronate also may increase bone proliferation and osteoblastic differentiation. However, little is known about mechanism of the action of alendronate on osteoblast differentiation, especially in transcription level. Inhibitors of DNA binding/differentiation (Ids) are helix-loop-helix (HLH) transcription factors and play an important role in BMP-induced osteoblast lineage-specific differentiation. Therefore, this study was aimed to investigate the effect of alendronate on osteoblast differentiation and expression of Id-1 and Id-2. Methods: MC3T3-E1, pre-osteoblast cell line, were treated with alendronate of various concentrations ($10^{-9}$ M-$10^{-4}$ M) and time periods (24, 48 and 72 hours). And then, the effect of alendronate on osteoblast differentiation was examined by alkaline phosphatase (ALP) activity and RT-PCR for osteoblast differentiation markers such as ALP, type 1 collagen (Col 1), and osteocalcin (OCN). The expressions of Id-1 and Id-2 were measured by RT-PCR. Results: Alendronate treatment increased not only ALP activity, but also expressions of ALP, Col 1, and OCN. Also, alendronate treatment up-regulated the mRNA levels of Id-1 and Id-2 genes. This alendronate-induced osteoblastic differentiation is more effective in lower doses rather than high doses. Conclusion: This study shows that the expression of transcription factor Id-1 and Id-2 was increased in a dose-dependent manner during alendronate-induced osteoblast differentiation.

목적: Alendronate는 대표적인 비스포스트네이트로서 파골세포의 활성을 떨어뜨려서 과도한 골흡수를 억제하는 약제로 알려져 왔으나, 최근에는 골세포 증식 및 분화를 증가시킨다고 보고되었다. 그러나 alendronate가 조골세포 분화에 미치는 기전에 대한 연구는 매우 부족하다. Id (Inhibitors of DNA binding/differentiation)은 helix-loop-helix (HLH) 계열에 속하는 전사인자이며, BMP에 의해 유도되는 골세포 분화과정동안 주요한 역할을 하는 것으로 알려져 있다. 그러므로 본 연구는 alendronate가 골세포 분화와 Id-1 및 Id-2 유전자의 발현에 미치는 효과를 조사하고자 한다. 연구 방법: 조골모세포인 MC3T3-E1 세포주에 alendronate를 다양한 농도 ($10^{-9}$ M-$10^{-4}$ M)와 시간 (24, 48 및 72시간)으로 처리한 후 조골세포로의 분화 정도를 ALP 활성도와 조골세포 분화표지자인 ALP, Col 1, OCN에 대한 발현을 역전사-중합효소반응 (RT-PCR) 방법으로 조사하였다. 또한, alendronate을 처리 후 Id-1 및 Id-2의 유전자 발현을 RT-PCR 방법으로 조사하였다. 결과: Alendronate는 ALP 활성도뿐만 아니라 ALP, Col 1 및 OCN의 mRNA 발현을 증가시키며, Id-1 및 Id-2의 발현도 증가시켰다. 이러한 alendronate에 의한 골세포 분화는 고농도보다 저농도에서 골세포 분화에 효과적이었다. 결론: 본 연구는 골형성과 관련된 전사인자 Id-1과 Id-2의 발현이 alendronate에 의한 조골세포 분화 과정동안 촉진됨을 보여주고 있으며, 이러한 alendronate 효과는 농도 의존적으로 골세포 분화에 영향을 미치는 것으로 사료된다.

Keywords

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