Co-Depletion of Cathepsin B and uPAR Induces G0/G1 Arrest in Glioma via FOXO3a Mediated p27Kip1 Upregulation
Authors: Jasti S Rao
Daniel Fassett
Jeffrey D Klopfenstein
Rama Rao Malla
Christopher S Gondi
Sreelatha Gopinath
Meena Gujrati
Kiranmai Alapati
Dzung H Dinh
Journal: PLoS ONE (2010)
Discipline: Multidisciplinary
Keywords: Cathepsin B; urokinase plasminogen activator receptor; gliomas; cell cycle progression; migration
Affiliations: Department of Cancer Biology and Pharmacology, University of Illinois College of Medicine at Peoria, Peoria, Illinois, United States of America.

Abstract:  Background: Cathepsin B and urokinase plasminogen activator receptor (uPAR) are both known to be overexpressed in gliomas. Our previous work and that of others strongly suggest a relationship between the infiltrative phenotype of glioma and the expression of cathepsin B and uPAR. Though their role in migration and adhesion are well studied the effect of these molecules on cell cycle progression has not been thoroughly examined. Methodology/principal findings: Cathepsin B and uPAR single and bicistronic siRNA plasmids were used to downregulate these molecules in SNB19 and U251 glioma cells. FACS analysis and BrdU incorporation assay demonstrated G0/G1 arrest and decreased proliferation with the treatments, respectively. Immunoblot and immunocyto analysis demonstrated increased expression of p27(Kip1) and its nuclear localization with the knockdown of cathepsin B and uPAR. These effects could be mediated by alphaVbeta3/PI3K/AKT/FOXO pathway as observed by the decreased alphaVbeta3 expression, PI3K and AKT phosphorylation accompanied by elevated FOXO3a levels. These results were further confirmed with the increased expression of p27(Kip1) and FOXO3a when treated with Ly294002 (10 microM) and increased luciferase expression with the siRNA and Ly294002 treatments when the FOXO binding promoter region of p27(Kip1) was used. Our treatment also reduced the expression of cyclin D1, cyclin D2, p-Rb and cyclin E while the expression of Cdk2 was unaffected. Of note, the Cdk2-cyclin E complex formation was reduced significantly. Conclusion/significance: Our study indicates that cathepsin B and uPAR knockdown induces G0/G1 arrest by modulating the PI3K/AKT signaling pathway and further increases expression of p27(Kip1) accompanied by the binding of FOXO3a to its promoter. Taken together, our findings provide molecular mechanism for the G0/G1 arrest induced by the downregulation of cathepsin B and uPAR in SNB19 and U251 glioma cells. 

Retracted: Yes

PubPeer comments:
 This  Mini Review article is co-authored by Dr. Sheikh, the current Editor-in-Chief of the journal. However, this author noted on [another thread] . This paper, as with multiple papers authored by Fleming, discussed in other threads on PubPeer, contains apparently re-used images. In this case, the figures appear to be cut and pasted with the captions of a previous paper(s) intact, as the footnote references do not match the footnotes in this article. As an example footnote [4] in the small print of Figure 1 refers in this paper to a 1994 publication by Fleming, Harrington, et. al. (doi.org 10.1177 000331979404501001) , that does *not* even contain Figure 1. ![file] First author Fleming responded to peer concerns about image reuse (including the image above) in this [thread] ,  The figures are copyrighted to the primary author and as such they remain the property of the primary author. I do not need to cite my own property. As Drs. Sheikh and Dooley have kindly responded in another thread, perhaps they would care to address the concerns about this paper as well. , 
 Several concerns about this paper. * Figure 3 appears to show a mammogram with a brief description of the patient, including age, gender, and possibly identifying medical information. There appears to be no statement of consent by this patient. * 11 of the 15 references (including #15, which refers to a patent written by the first author) are by the first author. See screenshot. ![file] ![file] , 
 As the authors did not provide a citation for their statements on the original purposes of the stress test, they may want to refresh their recollections. One source is here, which cites Bruceu2019s original paper: http: dx.doi.org 10.1136 heartjnl-2013-304003 ,