Silencing circular RNA circ_0010729 protects human cardiomyocytes from oxygen-glucose deprivation-induced injury by up-regulating microRNA-145-5p
Authors: Qifeng Jin
Yuanyuan Chen
Journal: Molecular and cellular biochemistry (2019)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: MEK/ERK pathway; MicroRNA-145-5p; Myocardial ischemia; Oxygen–glucose deprivation; circRNA_0010729
Affiliations: Department of Cardiology I, Affiliated Hospital of Jining Medical University, Jining, 272000, China., Department of Cardiology V, Affiliated Hospital of Jining Medical University, No. 89 Guhuai Road, Jining, 272000, Shandong, China. .

Abstract:  Circular RNAs (circRNAs) are effector molecules that exert functions in cardiovascular diseases. Nevertheless, the effects of circRNAs on myocardial ischemia remain uninvestigated. This paper aimed to explore the functions of circ_0010729 in oxygen-glucose-deprivation (OGD)-caused injury of human cardiomyocytes (HCM). HCM were exposed to OGD environment for 4 h. Then the expression of circ_0010729 was evaluated by RT-qPCR. After transfection, cell viability, apoptosis, and migration were examined to evaluate the impact of overexpression and knockdown of circ_0010729 on OGD-induced cell injury. The regulation between circ_0010729 and microRNA-145-5p (miR-145-5p) was verified. After miR-145-5p inhibitor transfection, whether aberrant miR-145-5p expression affected the modulation of circ_0010729 in OGD-induced cell injury was measured. Western blot was utilized to analyze mTOR and MEK/ERK pathway-related proteins. OGD treatment enhanced circ_0010729 expression and evoked cell injury in HCM. Moreover, OGD-induced injury was aggrandized by circ_0010729 overexpression via suppressing cell growth and migration in HCM. Knockdown of circ_0010729 attenuated OGD-induced injury. In addition, circ_0010729 negatively regulated miR-145-5p expression. MiR-145-5p inhibition reversed the effects of silencing circ_0010729 on OGD-induced injury and mTOR and MEK/ERK pathways. We demonstrated that silencing circ_0010729 activated mTOR and MEK/ERK pathways by up-regulating miR-145-5p, thereby protecting HCM from OGD-induced injury. 

Retracted: No

PubPeer comments:
 If I may plagiarise [Elisabeth M. Bik] : *The Western blots and other figures in this paper have an interesting layout that appears to be shared with many other papers, [...] The list of these papers is available here: https: docs.google.com spreadsheets d 1KXqTAyl4j-jVorFPMD2XRpr76LcIKJ0CVyIvRj0exYQ edit?usp=sharing* *In almost all papers, Western blot panels within the same figure, and across figures and papers, appear to share the same background, while the bands are regularly spaced, all have similar rounded edges without the usual smudges and specks, and with some bands showing a recognizable  jumping sardine shape.* *Also in almost all papers, bar graphs show a characteristic layout with bars in solid shades of grey, and the left most bar in black, as well as double-edged error bars. Many papers have  cell viability and  apoptotic cells graphs.* *In some of these papers, including this one, flow cytometry plot show a unnaturally dense and well defined  hairball of events in Q3 with a few regularly spaced events in Q1 and Q2.* In this case the FACS apoptosis plots are Figures 2B, 3C, 4D, 5E, 7C. Below, I have juxtaposed 3C *[upper]* with fig 3C *[lower]* from [Yi et al. (2019)] . A region of high overlap between two of the 3rd-Quadrant hairballs is marked. ![file] The overlap is especially surprising because of the lack of any connection between the authors or their institutions. , 
 #1 Here juxtaposing 3C from Yi et al. (2019) *[at left]* with Figs 2B, 3C from the present paper *[at right]*, to make that an entire panel is replicated between the two papers, differing only in the percentages cited for cell counts in each quadrant. ![file] , 
 #2 Here I annotated some of the duplicated panels, Q3 blobs or parts of Q3 blobs, replicated within or between Figures. I ran out of energy before considering 7C so that is left as an exercise for the reader. ![] , 
 And to add to Hoya s comments above, and to illustrate the recognizable layout of the graphs in this paper, here is Figure 2. ![file] , 
 In the western blots in the images shown by #4, the control Bax, HG cleaved caspase-3 and HG cleaved caspase-9 are identical much more similar that you would expect. There is also an obvious splicing line between Bax control and Bax HG. There could be others but these are just the immediate ones which stand out from figure posted by #4. , 
 #1 I have been examining a group of five papers, characterised by their distinctive Western Blots, which have a number of unifying features not in evidence elsewhere. The protein blobs have a distinctive appearance, vaguely resembling dog-bones or sometimes old-style telephone handpieces. Within each band the blobs are drawn from the same  morphological gamut , with no sense of development from one band to the next as the proteins migrate across an electrophoresis gel. The spaces between the lanes also remain constant from band to band, while the bands remain in perfect horizontal alignment, with no curvature or slant as they migrate (allowing for compactness when composing each Figure). Within a lane, there is no continuity from one band to the next, or between a protein and its phosphorylated form. I am not trying to show all of each paper s blots here, only a representative sample. 1. [Chen et al. (2018)] . Fig 5A: ![file] 2. [Li et al. (2019)] . Figs 5D, E, I, J. ![file] 3. [Lv et al (2019)] . Figs 4F, 4H. ![file] 4. [Yi et al. (2019)] . Figs 3D, 3G, 4B, 4C. ![file] 5. [Wan et al. (2020)] . Fig 7D: ![file] 4 and 5 also display some FACS plots (measuring cell apoptosis) of a distinctive (unique) appearance, described elsewhere as  The Death Star down in each frame s quadrant Q3 is shooting out bolts of planet-smashing energy to the right through Quadrant Q4, while disciplined flotillas of Rebel Alliance X-Wing starfighters are swooping down through Q1 and Q2 . These Death Star blots are also found in association with [the style of  leaping sardine Western blot] , and it may be that these airbrushed dog-bones are derived from the sardines. ![file] ![] To me it seems unlikely that these images were generated by five unrelated groups of authors, and I am more inclined to believe that they were the work of a single uncredited workshop. , 
 March 2020 Expression of Concern at https: www.tandfonline.com doi full 10.1080 21691401.2020.1741911 *After publication of this article, questions about the scientific integrity of the article content were brought to the Publisher and Editoru2019s attention. We have reached out to the authors requesting that they supply information that would confirm the articleu2019s integrity, but the authors have not responded to our queries within the requested timeframe. Therefore, as we continue to work through the issues raised, we advise readers to interpret the information presented in the article with due caution.* , 
 Retracted [21 October 2020] . -------------------------------------------------- We, the Editors and Publisher of the journal Artificial Cells, Nanomedicine, and Biotechnology, have retracted the following article: Wencui Wan, Weiwei Wan, Yang Long, Qiuming Li, Xuemin Jin, Guangming Wan, Fengyan Zhang Yong Lv. (2020). Physcion 8-O-u03b2-glucopyranoside exerts protective roles in high glucose-induced diabetic retinopathy via regulating lncRNA NORAD miR-125 STAT3 signalling. Artificial Cells, Nanomedicine, and Biotechnology, 48(1), 463u2013472, DOI: 10.1080 21691401.2019.1709861 This article has an expression of concern and there are 7 comments on PubPeer (by: Hoya Camphorifolia, Elisabeth M Bik, Myzobdella Lugubris) Since publication, concerns have been raised about the integrity of the data in the article. When approached for an explanation, the authors checked their data and confirmed there are fundamental errors associated with the figures presented. In addition, the corresponding author of the article, Qiuming, has informed us they were not aware of the submission made under their name. The authors have agreed to the retraction of this article, and we have also informed the institution about the authorship issues related to this article. We have been informed in our decision-making by our policy on publishing ethics and integrity and the COPE guidelines on retractions. The retracted article will remain online to maintain the scholarly record, but it will be digitally watermarked on each page as u2018Retractedu2019. ,