The silencing of long non-coding RNA ANRIL suppresses invasion, and promotes apoptosis of retinoblastoma cells through the ATM-E2F1 signaling pathway
Authors: Xiao-Wei Peng
Yang Yang
Journal: Bioscience Reports (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: ATM-E2F1 signaling pathway; Cell apoptosis; Cell proliferation; Long-non coding RNA ANRIL; Negative feedback regulation
Affiliations: Department of Pediatric Ophthalmology, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, P.R. China., Department of Pediatric Ophthalmology, The Second Affiliated Hospital of Nanchang University, Nanchang 330006, P.R. China .

Abstract:  As one of the most common primary intraocular carcinomas, retinoblastoma generally stems from the inactivation of the retinoblastoma RB1 gene in retinal cells. Antisense non-coding RNA in the INK4 locus (ANRIL), a long non-coding RNA (lncRNA), has been reported to affect tumorigenesis and progression of various cancers, including gastric cancer and non-small cell lung cancer. However, limited investigations emphasized the role of ANRIL in human retinoblastoma. Hence, the current study was intended to investigate the effects of ANRIL on the proliferation, apoptosis, and invasion of retinoblastoma HXO-RB44 and Y79 cells. The lentivirus-based packaging system was designed to aid the up-regulation of ANRIL and ATM expressions or employed for the down-regulation of ANRIL in human retinoblastoma cells. Afterward, ANRIL expression, mRNA and protein expression of ATM and E2F1, and protein expression of INK4b, INK4a, alternate reading frame (ARF), p53 and retinoblastoma protein (pRB) were determined in order to elucidate the regulation effect associated with ANRIL on the ATM-E2F1 signaling pathway. In addition, cell viability, apoptosis, and invasion were detected accordingly. The results indicated that the down-regulation of ANRIL or up-regulation of ATM led to an increase in the expressions of ATM, E2F1, INK4b, INK4a, ARF, p53, and pRB. The silencing of ANRIL or up-regulation of ATM exerted an inhibitory effect on the proliferation and invasion while improving the apoptosis of HXO-RB44 and Y79 cells. In conclusion, the key observations of our study demonstrated that ANRIL depletion could act to suppress retinoblastoma progression by activating the ATM-E2F1 signaling pathway. These results provide a potentially promising basis for the targetted intervention treatment of human retinoblastoma. 

Retracted: No

PubPeer comments:
 https: twitter.com BallouxFrancois status 1271195061540335616?s=20 , 
 #1 Rapid review by ncrc.jhsph.edu faculty recommending a retraction consideration: https: twitter.com KateGrabowski status 1271542361244352514?s=20 , 
 https: www.buzzfeednews.com amphtml peteraldhous mario-molina-coronavirus-face-masks-pnas , 
 https: twitter.com NoahHaber status 1273646681964208128 , 
 Direct link: https: metrics.stanford.edu PNAS%20retraction%20request%20LoE%20061820 , 
 The following letter was written and sent to the PNAS editorial board on July 18, 2020 to call for this paper to be retracted. The full letter with all signatories [is available here] . As of August 5, 2020 PNAS has not publicly responded to this call for retraction. Dear Editorial Board of the Proceedings of the National Academy of Sciences, We are writing with deep concerns about a paper recently published in your journal, entitled u201cIdentifying airborne transmission as the dominant route for the spread of COVID-19.u201d The paper made extraordinary claims about routes of transmission, the effectiveness of mask-wearing, and by implication, the ineffectiveness of other non-pharmaceutical interventions. While we agree that mask-wearing plays an important role in slowing the spread of COVID-19, the claims in this study were based on easily falsifiable claims and methodological design flaws. We present only a small selection of the most egregious errors here. Given the scope and severity of the issues we present, and the paperu2019s outsized and immediate public impact, we ask that the Editors of PNAS retract this paper immediately and reassess the Contributed Submission editorial process by which it was published. The main conclusions of this paper are based in comparison of linear case count slopes within and between regions, with mask mandates as the observed variable of interest. It ignores other clear differences in disease control policy between these areas, including broader heterogeneity in face mask policy. In one critical example, the paper asserts that u201cafter April 3, the only difference in regulatory measures between NYC and the United States lies in face coverings on April 17 in NYC.u201d This is verifiably false, based on widely available (e.g., HIT-COVID2) sources. It is flatly untrue that there were no other regulatory differences between NYC and the rest of the US on those dates; it is also untrue that NYC was the only region in the US mandating use of face coverings. In another example, the paper asserts that airborne transmission is the dominant route for COVID-19 spread. To justify this headline conclusion, the authors state that u201cWith social distancing, quarantine, and isolation in place worldwide and in the United States since the beginning of April, airborne transmission represents the only viable route for spreading the disease.u201d In fact, in April, many regions (e.g., Sweden, parts of the United States) were not in lockdown, and quarantine and isolation were not in place in most parts of the world. If similarly false statements about exposure were made widely in a cohort study or a randomized control trial, a rapid and complete retraction of the study would quickly follow. Hence, it is our view that PNAS is obligated to issue retraction of this work on these grounds alone. Beyond this, the study is afflicted with serious methodological errors that undermine any confidence in its findings. These include the following: 1) The analysis ignored the lag between changes in disease transmission and changes in reported case counts, 2) Dates of policy implementation are extremely poor proxies for mass behaviors, including social distancing and mask use, 3) Dates of policy implementation were concurrent with an enormous set of changes across society which plausibly affected reported incidence of COVID-19, 4) Case counts were modeled with a simple linear regressions, which is not consistent with infectious disease dynamics, 5) Demographics, policies, and contact behaviors in Wuhan, Italy, New York City, and the USA are inappropriately treated as being nearly equivalent to one another with respect to the epidemic, and 6) No measures of statistical uncertainty are measured or presented, which is a departure from scientific norms and particularly concerning given an analysis based on only three regions. Any one of the above issues in isolation would be cause for serious concern, but in combination, they are alarming. While masks are almost certainly an effective public health measure for preventing and slowing the spread of SARS-CoV-2, the claims presented in this study are dangerously misleading and lack any basis in evidence. Unfortunately, since its publication on June 11th, this article has been distributed and shared widely in traditional and social media, where its claims are being interpreted as 3 rigorous science. As societies debate the risks of re-opening and relaxing social distancing measures, it is crucial that decisions rely on a solid evidence base. The scale, scope, and intensity of the issues in this paper and the immediacy of the decisions being made based on its claimed conclusions do not make it amenable to either correction or extended debate within the academic literature . Accordingly, we ask that the editorial board of PNAS retract this paper immediately. Further, the scale of the methodological flaws raise concerns about the publication process. For this reason, we would argue that it may be time for PNAS to reconsider its policies on the Contributed Submissions track under which this paper was published, as this mechanism effectively bypasses editorial decisions and undermines peer review. While these may be painful processes, we believe that they constitute the best way forward in an extraordinary time for the scientific endeavor, and more importantly, for the health and well-being of the public. Noah Haber, ScD Postdoctoral Fellow Meta-Research Innovation Center at Stanford University noahhaber@stanford.edu Mary Kate Grabowski, PhD Assistant Professor Departments of Pathology and Epidemiology Johns Hopkins University mgrabow2@jhu.edu Daniel B. Larremore, PhD Department of Computer Science BioFrontiers Institute University of Colorado Boulder daniel.larremore@colorado.edu Nikolas Wada, PhD Co-lead, Non-Pharmaceutical Interventions Novel Coronavirus Research Compendium Johns Hopkins University nikwada@gmail.com Steven N. Goodman, MD, MHS, PhD Professor of Epidemiology and Population Health, and Medicine Stanford University steve.goodman@stanford.edu Justin Lessler Associate Professor Department of Epidemiology Johns Hopkins University justin@jhu.edu ,