MicroRNA-138 directly targets TNFAIP8 and acts as a tumor suppressor in osteosarcoma
Authors: Tao Chen
Zheng Zhou
Zhihong Li
Yi Shen
Journal: Experimental and therapeutic medicine (2017)
Discipline: CLINICAL MEDICINE
Keywords: microRNA; osteosarcoma; tumor necrosis factor-α-induced protein 8; tumor suppressor.
Affiliations: Department of Orthopaedics, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China.

Abstract:  MicroRNAs (miRs) have a critical role in the development and malignant progression of osteosarcoma (OS), but the underlying mechanisms have largely remained elusive. The present study aimed to explore the regulatory role of miR-138 in OS growth and metastasis and investigated the associated mechanisms. Reverse-transcription quantitative polymerase chain reaction and western blot analysis were performed to examine the miR-138 and protein expression levels in OS and normal bone tissues and cell lines. An MTT assay and a Transwell assay were used to assess cell proliferation and invasion. Flow cytometry was used to analyze the cell cycle and determine the apoptotic rate. A luciferase reporter assay was used to confirm the targeting association between miR-138 and tumor necrosis factor-α-induced protein 8 (TNFAIP8). It was found that miR-138 was downregulated in OS tissues and cell lines. Overexpression of miR-138 decreased the proliferation, cell cycle progression and invasion of OS cells, while inducing cell apoptosis. TNFAIP8 was then identified as a novel target of miR-138. Similarly to the effects of miR-138 overexpression, inhibition of TNFAIP8 also inhibited OS cell proliferation, cell cycle progression and invasion, and induced cell apoptosis. In addition, miR-138 overexpression as well as downregulation of TNFAIP8 reduced OS cell invasion via inhibition of matrix metalloproteinase-2 and -9 expression. Taken together, the results of the present study demonstrated that miR-138 directly targets TNFAIP8 and acts as a tumor suppressor in OS, suggesting that the miR-138/TNFAIP8 interaction may become a promising therapeutic target for OS. 

Retracted: Yes

PubPeer comments:
 Fig 5  Transwell assays were used to examine the invasion of Saos-2 and U2OS cells transfected with (A and B) miR-NC or miR-138 mimics and (C and D) NC siRNA or TNFAIP8u2011specific siRNA. ![file] I set up a spreadsheet to include the various papers using this versatile suite of Transwell images to represent all possible combinations of cell-lines and treatments. This is the 30th manifestation. For comparison: Fig 3 C D from [Zhang et al (2016)] .  Transwell assay were performed to examine the migratory and invasive capacities of Saos-2 and U2OS cells transfected with miR-205 mimic or miR-negative control (NC) mimic , and Fig 7C D,  Transwell assay were performed to examine cell migration and invasion of MG63 or U2OS cells transfected with miR-205 mimic, or co-transfected with miR-205 mimic and RUNX2 plasmid . ![] Figs 4D, 6D from [Xia et al (2016)] . ![] , 
 Dear Hoya Camphorifoliauff0c Thank you very much for your comments. In the process of the experiment, there may be some small mistakes in the pictures, and we are sure that this will not affect the results of the experiment. We will thoroughly check the original data and timely contact the journalu3002 Thank you very much again pointing out this question. Best regards , 
 #2  there may be some small mistakes in the pictures, and we are sure that this will not affect the results of the experiment The invasion-assay images illustrating one aspect of this paper have appeared in over **40 other papers**. How can this **not** affect the conclusions drawn from them? Consider the present Fig 4A,B:  Saos-2 and U2OS cells were transfected with NC siRNA or NFAIP8-specfic siRNA. (A and B) Western blot analysis was used to determine the protein levels of TNFAIP8 . ![file] Colored rectangles indicate surprisingly similar bands in Fig 7 from [Dai et al (2016)] .  Western blotting was performed to examine the (A) phosphorylated JNK (p-JNK) and total JNK (t-JNK), (B) phosphorylated ERK (p-ERK) and total ERK (t-ERK), (C) phosphorylated p38 (p-p38) MAPK and total p38 (t-p38) MAPK in the U87 cells transfected with the pcDNA3.1-CXCL5 plasmid or a blank vector as the negative control (NC), respectively. ![file] Fig 6A from [Wang et al (2017)] .  Western blot analysis was conducted to examine the expression of IGF2BP1 protein. ![file] *[above]* Figs 3H, 4A from [Zhou et al (2016a)] ; and [below] Figs 4C D from [Zhou et al (2016b)] .  Western blot was used to examine the protein levels of EMT-related genes . ![file] IN addition: Fig 6B from [Zhou et al (2017)] . Figs 7A,B, 9B from [Liu et al (2017)] . Figs 3B, 4B from [Dang et al (2017)] . Fig 4A from [Jiang et al (2017)] . ![file] , 
 Retracted [4 October 2021] .  Following the publication of this paper, it was drawn to the Editorsu2019 attention by a concerned reader that the Transwell cell migration assay data shown in Fig. 5Au2011D were strikingly similar to data appearing in different form in other articles by different authors. Owing to the fact that the contentious data in the above article had already been published elsewhere, or were already under consideration for publication, prior to its submission to *Experimental and Therapeutic Medicine*, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive any reply. The Editor apologizes to the readership for any inconvenience caused. ,