MicroRNA-27a-3p Modulates the Wnt/β-Catenin Signaling Pathway to Promote Epithelial-Mesenchymal Transition in Oral Squamous Carcinoma Stem Cells by Targeting SFRP1
Authors: Jing-Hua Cai
Alfred King-Yin Lam
Qian Tao
Bao-Xia He
Bin Qiao
Journal: Scientific Reports (2017)
Discipline: Multidisciplinary
Keywords: microRNA27a-3p; Wnt/β-catenin signaling pathway; epithelial-mesenchymal transition (EMT); oral squamous carcinoma stem cells (OSCSCs); secreted frizzled-related protein 1 (SFRP1)
Affiliations: Department of Stomatology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, P. R. China., Department of Oral and Maxillofacial Surgery, Guanghua School and Hospital of Stomatology; Guangdong Provincial Key Laboratory of Oral Diseases, Sun Yat-sen University, Guangzhou 510055, P. R. China., Department of Pharmacy, Henan Cancer Hospital/Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou, 450052, P. R. China., Cancer Molecular Pathology, School of Medicine and Menzies Health Institute Queensland, Griffith University, Gold Coast, 4222, Australia.

Abstract:  This study aimed to elucidate how microRNA27a-3p (miR-27a-3p) modulates the Wnt/β-catenin signaling pathway to promote the epithelial-mesenchymal transition (EMT) in oral squamous carcinoma stem cells (OSCSCs) by targeting secreted frizzled-related protein 1 (SFRP1). Flow cytometry was used to sort OSCSCs from the SCC-9 and Tca8113 cell lines. The OSCSCs were randomly assigned into the miR-27a-3p inhibitors group, the miR-27a-3p inhibitors-NC group, the si-SFRP1 group, the si-SFRP1 + miR-27a-3p inhibitors group and the blank group. A luciferase reporter, immunofluorescence and Transwell assays were performed to detect luciferase activity, SFRP1, and cell migration and invasion, respectively. The mRNA expression of miR-27a-3p, SFRP1 and EMT markers (E-cadherin, N-cadherin, vimentin and ZEB1) were detected using qRT-PCR. The protein expression of SFRP1, EMT markers and the proteins of the Wnt/β-catenin signaling pathway was detected by Western blotting. OSCSCs showed up-regulated miR-27a-3p, Wnt/β-catenin signaling pathway-related proteins, vimentin, N-cadherin and ZEB1 and down-regulated SFRP1 and E-cadherin. MiR-27a-3p targeted SFRP1. Down-regulated miR-27a-3p resulted in increased E-cadherin and SFRP1 but decreased vimentin, N-cadherin, ZEB1, the Wnt/β-catenin signaling pathway-related proteins, and invasive and migratory cells. Silenced SFRP1 reversed this effect. We found that miR-27a-3p modulated the Wnt/β-catenin signaling pathway to promote EMT in OSCSCs by down-regulating SFRP1. 

Retracted: Yes

PubPeer comments:
 Figure 8 appears to have overlapping images for different treatments and cell types. ![file] Additionally, there is also overlap between figure 8 and figure 9 (light green box). Figure 9 also has other overlapping images. ![file] , 
 There also appears to be duplication of blots in figure 4, even though they are on different cell lines. ![file] And the same appears true for figure 5. ![file] And again for figure 7. ![file] , 
 There also appear to be the same or similar areas in figure 1 flow cytometry plots of different cell lines. However, I am not very familiar with flow cytometry so perhaps that is normal. ![file] , 
 Additionally, I ve realized this paper appears to share images with another paper (doi: 10.1159 000479610 , [see pubpeer comment] ) The other paper is also about miR-27a and SFRP1, but in colon cancer cells, not oral squamous carcinoma stem cells. Most images have the same orientation in both figures, but one appears to have been rotated (yellow box). This paper s figure 9: ![file] The other paper s relevant figure: ![file] , 
 #3  There also appear to be the same or similar areas in figure 1 flow cytometry plots of different cell lines. The vertical shift in those areas between the two images rules out any explanation in terms of the same data set being accidentally plotted twice (with different filter parameters). , 
 #4 Thank you very much for your comments. My team is contacting the editorial office to deal with this issueu3002 , 
 #4 Here repeating Dendrodoa Grossularia s observations about the links between the present Figs 8 and 9 (at left) with Fig 5 from [Ba et al (2017)] (upper right), but I have added Fig 5B from [Xie et al (2017)] (lower right). I have not tried to fit the links **within** Figs 8 and 9, for that would need too many colours. ![file] , 
 I agree with the first and corresponding author comments as well as the comments of in this group The first and corresponding author will investigate in his team and have contacted the journal for the article , 
 Some more overlaps found with[ this paper] from 2015: ![file] , 
 Fig 8 again: ![file] Panels rotated and color-coded to facilitate comparison with FIG. 11 from [He et al (2016)] .  Migration ability of U2OS cells were detected by Transwell experiment. (A) Blank group; (B) Scramble group; (C) miR-106a-5p group; (D) HMGA2-siRNA group; (E) miR-106a-5p+HMGA2 group . ![file] Fig 9B, again with panels rotated and color-coded... ![] ...for comparison with Fig 12 from He et al. ![] . , 
 #10 Thank you very much for the question you pointed out. My students was in charged of the part of the Transwell experiment and the lab responsible for this result. As the students lack of experience, the stored names of the groups after taking pictures are not standardized and the layout is negligent, which leads to the situation that the pictures are overlapped, and the confusion of the experimental data and picture management leads to the possible picture leakage. I attached great importance to this issue. We are verifying the experimental results and contacted the editor of the journal magazine to deal with this problem. We realized the that the result may be not rigorous, and I am negotiating with the editor in chief about the retraction of the manuscript. Thanks for your patience and we will handle it as soon as possible. , 
 #11 I can see how a student s inexperience with image organisation might have led to the overlaps between images in Figures 8 and 9. However, I don t see how it could account for the appearance of the same images in a number of other papers, published by unrelated teams of researchers, both previously and subsequently to the present paper. , 
 Retracted on December 1st, 2020 https: www.nature.com articles s41598-020-77203-x  Retraction of: Scientific Reports https: doi.org 10.1038 srep44688, published online 20 April 2017 The authors have retracted this Article. After publication they became aware that the oral cancer cell line had been contaminated with HeLa cells. In addition, there are irregularities in a number of figures, specifically: * Apparent overlap of the flow cytometry plots shown in Figure 1A and 1B which are for different cell lines * Duplication of blots between Figure 4A and Figure 4C * Duplication of blots between Figure 5B and Figure 5D * Duplication of blots between Figure 7B and Figure 7D * Duplication of parts of panels both within Figures 8 and 9 and between Figures 8 and 9 and parts of panels in figures in other articles [1](http: www.karger.com Article FullText 479610), [2](http: doi.org 10.1042 BSR20160542), [3](http: www.nature.com articles cddis2017561), [4](http: www.karger.com Article Abstract 437379) All authors agree to this retraction , 
 **Decision on handling misconduct cases investigated and handled in 2021 (third batch)** https: www.nsfc.gov.cn publish portal0 tab434 info81957.htm This paper is discussed among the 22 handling decisions announced by the National Natural Science Foundation of China u201cthat have been criticized for notification are now published.u201d (Google Translate). Regarding the tampering of pictures and unauthorized Use other people s signatures and mark other people s fund projects without authorization And other issues National Science and Technology Supervision Office (2021) No. 82 u3000u3000 u3000u3000The Supervision Committee of the National Natural Science Foundation of China reviewed the retracted paper  Bin Qiao # *, Bao-Xia He # , Jing-Hua Cai, Qian Tao, Alfred King-yin Lam*. MicroRNA-27a-3p Modulates the Wnt u03b2-Catenin Signaling Pathway to Promote Epithelial-Mesenchymal Transition in Oral Squamous Carcinoma Stem Cells by Targeting SFRP1. Scientific Reports, 2017, 7:44688. (labeled with fund numbers 81200796, 81302796), an investigation was launched for suspected academic misconduct. u3000u3000After investigation, the paper has the problems of tampering with pictures, using other people s signatures without permission, and unauthorized labeling of other people s National Natural Science Foundation projects. The first and corresponding author Qiao Bin bears full responsibility. u3000u3000After deliberation at the 10th meeting of the 5th Supervisory Committee of the National Natural Science Foundation of China (the Professional Committee of Life Medicine), the National Natural Science Foundation of Handling Measures Article 47, Article 40, Article 43, Item 1 and 6, cancel the Qiaobin National Natural Science Foundation project  Three-dimensional culture and transduction of Sox2 induced human buccal mucosal epithelial stem cells into iPSC Researchu201d (Approval No. 81200796), recover the allocated funds, cancel Qiao Bin s National Natural Science Foundation project application and participation qualifications for 3 years (July 20, 2021 to July 19, 2024), give Qiao Bin a circular and criticize . , 
 #14  using other people s signatures without permission It sounds as if some co-authors belatedly decided that they weren t co-authors after all. There is also the involvement of [a papermill] , unmentioned in the report. ,