Retracted : MicroRNA‐539 promotes osteoblast proliferation and differentiation and osteoclast apoptosis through the AXNA‐dependent Wnt signaling pathway in osteoporotic rats
Authors: Zheng-Xiang Huang
Wen-Jun Lin
Chen Lv
Xin Chen
Lu Wang
Xiong-Bai Zhu
Sheng-Wu Yang
Journal: Journal of Cellular Biochemistry (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: axis inhibition protein 1; microRNA-539; osteoblast; osteoclast; osteoporosis
Affiliations: Department of Orthopedics, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, P.R. China.

Abstract:  This study aims to explore the effects of miR-539 on osteoblast proliferation and differentiation and osteoclast apoptosis in a rat model of osteoporosis, and its mechanism involving the regulation of the AXIN1-mediated wingless-Int (Wnt) signaling pathway. A rat model of osteoporosis was successfully established by ovariectomy. With osteoblasts and osteoclasts of rats not receiving ovariectomy in the sham group as control, those of osteoporotic rats were treated with miR-539 inhibitor, miR-539 mimic, and AXIN1 shRNA. The expression of miR-53, AXIN1, the Wnt pathway related-genes, apoptosis related-genes, and osteogenic markers were measured by RT-qPCR and Western blot analysis, respectively. Alkaline phosphatase (ALP) activity in osteoblast and tartrate-resistant acid phosphatase (TRAP) activity in osteoclasts were determined after cell transfection. Osteoblast and osteoclast viability was assayed by CCK-8 assay. Cell cycle and apoptosis of osteoblasts and osteoclasts were detected by flow cytometry. Lastly, alizarin red S staining was used to detect mineralized nodules of osteoblasts. Firstly, we determined that miR-539 was down-regulated in osteoblast and osteoclast of osteoporotic rats and AXIN1 was negatively regulated by miR-539. Additionally, overexpression of miR-539 increased the expressions of β-catenin, LEF1, c-myc, cyclin D1, RUNX2, BGP, BMP-2 in osteoblast as well as β-catenin, RhoA, caspase-3, and Bcl-2 in osteoclasts. Finally, overexpression of miR-539 elevated ALP activity, proliferation, and mineralized nodules in osteoblast and osteoclast apoptosis, with reduced TRAP activity in osteoclasts. Our results demonstrate that miR-539 promotes osteoblast proliferation and differentiation as well as osteoclast apoptosis through the AXIN1-dependent Wnt signaling pathway in osteoporotic rats. 

Retracted: Yes

PubPeer comments:
 FIGURE 2B  Relative mRNA and protein expression of the Wnt pathway-related genes in osteoblasts and osteoclasts after transfection using RT-qPCR and Western blot analysis.... (B) relative protein expression of the Wnt pathway-related genes in osteoblasts . ![file] The loading control has appeared in a number of otherwise-unrelated papers. Fig 5B from [Wu et al (2018)] : ![] FIGURE 5B from [Wang et al (2018)] and Figure 6A,C from [Wen et al (2018)] . ![] , 
 FIGURE 3B  Relative mRNA and protein expression of osteogenic markers and apoptosis-related genes in osteoclasts after transfection using RT-qPCR and Western blot analysis. ... (B) relative protein expression of osteogenic markers . ![file] Another familiar loading control. E.g. Fig 9A from [Zhou et al (2018)] . ![] Fig 3A from [Gong et al (2018)]. ![] Other examples at the Zhou et al thread. , 
 Fig 3D. Again, the loading band is like an old friend. ![file] , 
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 Retracted [30 June 2021] . --------------------------------- The above article, published online on 12 June 2018 in Wiley Online Library has been retracted by agreement between the journal s Editor in Chief, Prof. Dr. Christian Behl, and Wiley Periodicals LLC. The retraction has been agreed following an investigation based on allegations raised by a third party. A detailed investigation revealed that several image elements of the experimental data were published elsewhere in a different scientific context. Thus, the editors consider the conclusions of this article to be invalid. ,