MiR-124 regulates osteoblast differentiation through GSK-3β in ankylosing spondylitis
Authors: Q-H Huang
L-G Wu
C Liu
S-L Tang
A-L Cai
Journal: European review for medical and pharmacological sciences (2018)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: Ankylosing spondylitis; miR-124; GSK-3β; Wnt/β-catenin pathway; osteoblast differentiation
Affiliations: Department of Spinal Surgery, Zhuzhou Central Hospital, Zhuzhou, Hunan, China. .

Abstract:  Objective: Ankylosing spondylitis (AS) is a spastic and spinal joint disease with the characteristic of pathological ossification. Bioinformatics analysis demonstrated that there is a complementary binding site between microRNA-124 (miR-124) and the 3'-UTR of glycogen synthase kinase-3β (GSK-3β) mRNA. We aimed to investigate the role of miR-124 in regulating GSK-3β expression, Wnt/β-catenin pathway activity, and osteoblast differentiation of spinal ligament fibroblasts. Patients and methods: The ligament tissues of AS and the femoral neck fracture patients were collected. MiR-124 and GSK-3β mRNA expressions were detected by using quantitative Real-time PCR (qRT-PCR). GSK-3β and β-catenin protein expressions were detected by using Western blot. Ligament fibroblasts were isolated and induced to differentiate into osteoblasts. Alizarin red S staining (ARS) was used to identify osteoblast differentiation. Expressions of miR-124, GSK-3β, β-catenin, Osterix, and runt-related transcription factor 2 (RUNX2) were detected during differentiation. The cells were divided into two groups as agomiR-normal control (NC) transfection group and agomir miR-124 transfection group. Alkaline phosphatase (ALP) activity and Alizarin Red S staining were detected. Results: MiR-124 and β-catenin expressions in the ligament of AS patients increased, while GSK-3β level reduced compared with control. MiR-124, β-catenin, Osterix, and RUNX2 expressions gradually elevated, whereas GSK-3β level gradually declined following increased osteoblasts differentiation. Antagomir miR-124 transfection significantly up-regulated the expression of GSK-3β in osteoblast differentiation, significantly decreased the expression of β-catenin, Osterix, and RUNX2, and significantly inhibited osteoblast differentiation. Conclusions: MiR-124 decreased and GSK-3β elevated in AS ligament tissue. Down-regulation of miR-124 expression enhanced GSK-3β expression, weakened Wnt/β-catenin pathway activity, and inhibited the differentiation of ligament fibroblasts into osteoblasts. 

Retracted: No

PubPeer comments:
 This article makes a methodological mistake that renders its conclusions invalid. As the article explains:  Among applicants who were admitted and matriculated into physics Ph.D. programs, we find the predictive validity to be poor for some of the most ubiquitously used admissions criteria. The methodological mistake is called restriction of range. Because a Ph.D. programme will only admit applicants with high GRE scores, the correlation between GRE scores and programme completion is necessarily attenuated because the correlation is estimated solely based on the data of people with high GRE scores. This is the same reason why no correlation exists between height and performance in professional American basketball. It is not because height does not matter for performance in professional basketball. It is because only tall people are admitted into the basketball league at the professional level. A restriction in range of the data attenuates the correlation and thus prevents one from concluding that there is no correlation between height and performance. That the authors make no mention of this fatal flaw is surprising and that the editorial team failed to identify it either is disappointing and casts doubt on the rigour of Science Advances. Without belabouring the point or criticising other limitations (such as the selection of mere programme completion as the dependent variable, rather than publication record or some other more ecologically valid variable), the conclusions of this study are invalid and certainly should not be relied upon to inform public policy or graduate programme admissions procedures. , 
 #1 The problem is worse than that, because they post-stratified the students by the ranking of the program in which they enrolled. Thus they greatly amplified the range restriction problem that most studies have. There are many other extremely serious problems, including failing to deal properly with collinearity, misinterpreting the meaning of failure to reject a null, and multiplying some confidence intervals by a factor of ten to obscure trends. , 
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