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Vol 52, No 3 (2014)
Original paper
Submitted: 2014-05-25
Accepted: 2014-09-09
Published online: 2014-10-10
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GABPβ2 expression during osteogenic differentiation from human osteoblast-like Saos-2 cells

Xiaoyuan Xu, Jianjun Xiong, Tao Wang, Meirong Zheng, Ping Wu, Xinping Wang, He Jiang, Benyi Yi, Bin Lang, Weidong Li
DOI: 10.5603/FHC.2014.0026
·
Folia Histochem Cytobiol 2014;52(3):225-231.

open access

Vol 52, No 3 (2014)
ORIGINAL PAPERS
Submitted: 2014-05-25
Accepted: 2014-09-09
Published online: 2014-10-10

Abstract

The E26 transformation-specific (ETS) family of transcription factors plays an important role in osteogenic differentiation. Whether GA-binding protein β2 (GABPβ2), a member of the ETS family, is involved in osteogenic differentiation has not been previously reported. In the present study, directed differentiation of human osteoblast-like Saos-2 cells was induced and validated by examining alkaline phosphatase (ALP) activity, presence of mineralized nodule and other phenotypic characteristics of the cells on days 0, 3, 6 and 9, thus establishing their osteogenic potential. Real-time PCR revealed that similarly to the bone-specific transcription factor Runx2, the expression of Gabpb2 in Saos-2 cells also peaked on day 3 and was significantly reduced on days 6 and 9. Immunocytochemical staining showed that changes in the immunoreactivity of GABPβ2 also exhibited a similar trend to that of Runx2. Initially, Runx2 was predominantly localized in the nuclei, while GABPβ2 was relatively diffuse. Both exhibited a significant increase in immunoreactivity on day 3, with presence in both the nuclei and cytoplasm. By day 6, both showed a significant decrease in immunoreactivity and were mainly localized in the nuclei. Therefore, we surmise that GABPβ2, as an ETS family member, may play a regulatory role in early osteoblastic differentiation and potentially act in synergy with Runx2.

Abstract

The E26 transformation-specific (ETS) family of transcription factors plays an important role in osteogenic differentiation. Whether GA-binding protein β2 (GABPβ2), a member of the ETS family, is involved in osteogenic differentiation has not been previously reported. In the present study, directed differentiation of human osteoblast-like Saos-2 cells was induced and validated by examining alkaline phosphatase (ALP) activity, presence of mineralized nodule and other phenotypic characteristics of the cells on days 0, 3, 6 and 9, thus establishing their osteogenic potential. Real-time PCR revealed that similarly to the bone-specific transcription factor Runx2, the expression of Gabpb2 in Saos-2 cells also peaked on day 3 and was significantly reduced on days 6 and 9. Immunocytochemical staining showed that changes in the immunoreactivity of GABPβ2 also exhibited a similar trend to that of Runx2. Initially, Runx2 was predominantly localized in the nuclei, while GABPβ2 was relatively diffuse. Both exhibited a significant increase in immunoreactivity on day 3, with presence in both the nuclei and cytoplasm. By day 6, both showed a significant decrease in immunoreactivity and were mainly localized in the nuclei. Therefore, we surmise that GABPβ2, as an ETS family member, may play a regulatory role in early osteoblastic differentiation and potentially act in synergy with Runx2.

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Keywords

GABPβ2; Runx2; human osteoblast-like Saos-2 cells; osteogenic differentiation; IHC; QPCR

About this article
Title

GABPβ2 expression during osteogenic differentiation from human osteoblast-like Saos-2 cells

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 52, No 3 (2014)

Article type

Original paper

Pages

225-231

Published online

2014-10-10

Page views

1881

Article views/downloads

2607

DOI

10.5603/FHC.2014.0026

Bibliographic record

Folia Histochem Cytobiol 2014;52(3):225-231.

Keywords

GABPβ2
Runx2
human osteoblast-like Saos-2 cells
osteogenic differentiation
IHC
QPCR

Authors

Xiaoyuan Xu
Jianjun Xiong
Tao Wang
Meirong Zheng
Ping Wu
Xinping Wang
He Jiang
Benyi Yi
Bin Lang
Weidong Li

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