microRNA-145 inhibits osteosarcoma cell proliferation and invasion by targeting ROCK1
Authors: Jie Xie
Yihe Hu
Long Wang
Pengfei Lei
Zixun Dai
Xucheng Yang
Journal: Molecular Medicine Reports (2014)
Discipline: CLINICAL MEDICINE
Keywords: Osteosarcoma; microRNA-145; ROCK1; cell proliferation; invasion
Affiliations: Department of Orthopedics, Xiangya Hospital of Central South University, Changsha, Hunan 410008, P.R. China.

Abstract:  Osteosarcoma (OS), a malignant mesenchymal sarcoma, is the most frequent primary bone tumor, with a peak incidence in young children and adolescents. The downregulation of microRNA‑145 (miRNA/miR‑145) has previously been identified to be associated with the aggressiveness and metastasis of OS. However, the detailed regulatory mechanism by which miR‑145 inhibits OS remains largely unknown. The present study demonstrated that miR‑145 was significantly downregulated in OS tissues and KHOS and U2OS cell lines. Rho‑associated protein kinase 1 (ROCK1), a key regulator of actin cytoskeleton reorganization, was identified as a novel target of miR‑145. Ectopic expression of miR‑145 notably suppressed the protein expression of ROCK1 without affecting its mRNA level. Furthermore, the expression of ROCK1 was significantly increased in the OS tissues and in the KHOS and U2OS cells. It was further demonstrated that the overexpression of miR‑145 downregulated KHOS and U2OS cell proliferation and invasion, which was reversed by restoration of ROCK1. To the best of our knowledge, the present study demonstrates for the first time that, as a tumor suppressor, miRNA‑145 inhibits OS cell proliferation and invasion, at least in part by directly targeting ROCK1. These results indicate that miR‑145 may be a potential candidate for the diagnosis and treatment of OS. 

Retracted: Yes

PubPeer comments:
 Fig 3A.  Western blotting data demonstrating that co-transfection with miR-145 and ROCK1 attenuated the suppressive effect of miR-145 on ROCK1 protein expression. ![file] Flipped horizontally, one band is unexpectedly similar to bands in Figs 5 and 3C of [Tan et al (2016)] . ![] , 
 *[below]* Fig 4B.  Western blotting data revealing that miR-145 significantly inhibited the protein expression of MMP2 and MMP9 in KHOS and U2OS cell invasion, which was reversed by ROCK1 overexpression . *[above[* Fig 4A from [Xiao et al (2016)] .  Western blotting was performed to examine the protein level of DLCu20111 in H1229 cells transfected with miRu2011429 mimic, or cou2011transfected with miRu2011429 mimic and DLCu20111 plasmid, respectively . Unexpected similarity of two bands (after 180-deg flip). ![file] , 
 Fig 4A. ![file] For comparison: Fig 5A of [Long et al (2016)] .  A Transwell assay was performed to determine the invasion capacity of T24 cells following 24 or 48 h of transfection with miR-101 mimics, c-FOS siRNA, or co-transfected with miR-101 mimics and c-FOS plasmid. (A) Representative images of lower sides of the Transwell membranes with invaded cells stained with crystal violet. ![] Figs 2D, 4C of [Ma et al (2017)] . ![] UPDATE: Figs 3b, 7b of [Zhong et al (2018)] . ![file] , 
 Dear Hoya: Thanks for your comments. We will investigate this and I discussed with my team members about repeating these years-ago experiments because these experiments in a public laboratory that shared with many other researchers. , 
 #4 Thank you for responding. While it may be interesting to see the results of new experiments, can you explain how that would address the concerns with the provenance of the data in this already published paper? Have you contacted the journal to hear how they would like to proceed? , 
 If it would help, I can list the 11 other papers using the same suite of crystal-violet-stained Transwell invasion results to illustrate different combinations of cell-lines and treatments. , 
 #3 The **internal** overlaps within Fig 4 are more extensive than I noted earlier. ![file] , 
 A coincidence? Figure 4B, Molecular Medicine Reports (2014), doi: 10.3892 mmr.2014.2195, Received September 4, 2013 Figure 5A, Scientific Reports (2013), doi: 10.1038 srep03423, Received September 17, 2013 *** ![file] , 
 #8 ![file] Row 14 in this spreadsheet: https: docs.google.com spreadsheets d 1I7uuhZ7NgMtzXFaJVfBPzP92JjCHO8tJDOjL8Y_9q1U edit?usp=sharing , 
 Retracted [4 July 2022] .  Following the publication of this paper, it was drawn to the Editorsu2019 attention by a concerned reader that several of the data panels shown in Fig. 4A were overlapping with panels contained within the same figure, where the panels were intended to portray the results from a range of different cell migration assay experiments. Given the number of overlaps of data that have been identified, the Editor of Molecular Medicine Reports has decided that this paper should be retracted from the Journal on account of a lack of confidence in the presented data. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a satisfactory reply. The Editor apologizes to the readership for any inconvenience caused. ,