Inhibition of microRNA‐940 suppresses the migration and invasion of human osteosarcoma cells through the secreted frizzled‐related protein 1–mediated Wnt/β‐catenin signaling pathway
Authors: Zun-Wen Lin
Xiao-Feng Li
Wen-Bo Wei
Wei Zhang
Si-Ding Jiang
Journal: Journal of Cellular Biochemistry (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Wnt/β-catenin signaling pathway; apoptosis; invasion; microRNA-940 (miR-940); migration
Affiliations: Department of Orthopedics, The First Affiliated Hospital of Nanchang University, Nanchang, China., Department of Respiratory Medicine, The First Affiliated Hospital of Nanchang University, Nanchang, China.

Abstract:  Osteosarcoma (OS) is the most common malignant tumor of bone with a high potential for metastasis. This study intends to explore whether microRNA-940 (miR-940) affects the development of OS cells and the underlying mechanism. OS and adjacent normal tissues were collected from OS patients; the OS cell line with the highest expression of miR-940 was selected, which was then subjected to transfection of miR-940 mimic, miR-940 inhibitor, siRNA-secreted frizzled-related protein 1 (SFRP1) or LiCl (agonists of Wnt/β-catenin pathway) to identify regulation of miR-940 to OS cells through SFRP1. The targeting relationship between miR-940 and SFRP1 was verified using dual-luciferase reporter gene assay. Reverse-transcription quantitative polymerase chain reaction and Western blot assay were performed to determine miR-940, SFRP1, β-catenin, and cyclinD1 and apoptosis-related genes Fas, Bax, and Bcl-2. MTT (3-(4, 5-dimethylthiazol-2-Yl)-2, 5-diphenyltetrazolium bromide) assay, scratch test, transwell assay, and flow cytometry were carried out to detect proliferation, migration, invasion, and apoptosis, respectively. Nude mice models were established to observe the tumor formation. Higher expression of miR-940, β-catenin, and cyclinD1 and lower SFRP1 expression were identified in OS tissues. miR-940 targeted and negatively regulated SFRP1 expression. Furthermore, upregulated miR-940 expression activated the Wnt/β-catenin signaling pathway in OS. With the treatment of miR-940 mimic, LiCL, or siRNA-SFRP1, OS cells showed promoted proliferation, migration, invasion, tumor formation, and impeded apoptosis (further reflected by elevated Bcl-2 expression and reduced Fas and Bax expression). The study demonstrates that miR-940 can promote the proliferation, migration, and invasion but suppress the apoptosis of human OS cells by downregulating SFRP1 through activating Wnt/β-catenin signaling pathway. 

Retracted: Yes

PubPeer comments:
  FIGURE 5 Upregulated miRu2010940 expression reduced protein expression of SFRP1 expression and the Wnt u03b2u2010catenin signaling pathwayu2010 related genes in OS cells. A, protein bands of SFRP1, u03b2u2010catenin, and cyclinD1 in U2OS cells . Below: Figs 9A from [Y.-H. Wu et al (2018)] and 5C from [D.-M. Wu et al (2018)] . Protein blobs seem to come from the same restricted repertoire. In particular, there are remarkable similarities within the loading controls. ![file] , 
 Fig 9A(e). The idea with these flow-cytometry diagrams is that the smooth black line is supposed to be the sum of the red peaks and the striped hump, which have been scaled so that their sum best approximates the empirical cell-count histogram (jagged black line). If the line is **not** the sum of those scaled templates, we have to conclude that the graph was hand-drawn. ![file] , 
 Retracted [19 July 2021] . ---------------------------------------------- The above article, published online on 15 October 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. ,