RETRACTED ARTICLE: Chemopreventive Potential of Resveratrol in Mouse Skin Tumors Through Regulation of Mitochondrial and PI3K/AKT Signaling Pathways
Authors: Preeti Roy
Jasmine George
Sahdeo Prasad
Neetu Kalra
Yogeshwer Shukla
Journal: Pharmaceutical Research (2009)
Discipline: PHARMACOLOGY & TOXICOLOGY
Keywords: Resveratrol; Chemoprevention; Tumorigenesis; Apoptosis; Skin tumors
Affiliations: Proteomics Laboratory, Indian Institute of Toxicology Research, (Council of Scientific & Industrial Research, India), P.O. Box 80, M.G. Marg, Lucknow, 226001, India.

Abstract:  Purpose: To investigate the chemopreventive potential of resveratrol, a phytoalexin found in seeds and skin of grapes, berries and peanuts in 7,12 dimethyl benz(a)anthracene (DMBA) induced mouse skin tumorigenesis. Methods: Topical treatment of resveratrol was given to the animals 1 h prior to DMBA for 28 weeks. At the end of the study period, the skin tumors were dissected out and western blotting was carried out to examine the regulation of proteins involved in anti-tumorigenesis in response to resveratrol. Results: Chemopreventive properties of resveratrol were reflected by delay in onset of tumorigenesis, reduced cumulative number of tumors, and reduction in tumor volume. Results of the western blotting showed that resveratrol treatment increased the DMBA suppressed p53 and Bax while decreased the expression of Bcl-2 and Survivin. Further, resveratrol supplementation resulted in release of cytochrome C, caspases activation, increase in apoptotic protease-activating factor-1 (Apaf-1) as mechanism of apoptosis induction. Resveratrol was also found to inhibit skin tumorigenesis through regulation of Phosphatidylinositol-3-kinase (PI3K)/ and AKT proteins which are implicated in cancer progression because it stimulates proliferation and suppresses apoptosis. Conclusions: Based on the results we can conclude that resveratrol regulates apoptosis and cell survival in mouse skin tumors as mechanism of chemoprevention hence deserve to be a chemopreventive agent. 

Retracted: Yes

PubPeer comments:
 One set of b-actin W.B. has been recycled in as many as five different papers between 2006-2009. DOI: 10.1016 j.canlet.2005.10.049 Fig 1: Mouse skin DOI:10.1002 mnfr.200700197 Fig 6 (F): Effect of gingerol on prostate cancer cells DOI: 10.1007 s11095-008-9723-z Fig 3 (B): Effect of resveratrol on mouse skin tumor doi:10.1016 j.lfs.2007.04.009 Fig 3 (A): LNCaP cells DOI: 10.1080 01635580701613772 Fig 8 (B): Lymph node carcinoma of the prostate cells tissue ![file] ![file] , 
  Fig. 3. Western blots showing the effect of resveratrol on a p53, b Bax, c Bcl-2, **d cytochrome C**, e Apaf-1 in mouse skin tumors in different groups.  ![file] The beta-Actin band cries out for an explanation. , 
  Fig. 3. Western blots showing the effect of resveratrol on a p53, b Bax, **c Bcl-2**, d cytochrome C, e Apaf-1 in mouse skin tumors in different groups.  ![file] An identical beta-Actin band appears in Figure 1(B),  Western blot analysis of mutp53 in mouse skin tumors following DAS application from [Arora c (2004)] , despite completely different conditions. , 
  Fig. 3. Western blots showing the effect of resveratrol on **a p53**, b Bax, c Bcl-2, d cytochrome C, e Apaf-1 in mouse skin tumors in different groups.  ![file] The p53 band seems to have been pasted together like a stamp collection. As for the beta-actin control, it has appeared in [at least two other experiments] . ![] , 
 #1  Fig. 4. Western blots showing the effect of resveratrol on [...] **c PARP** in mouse skin tumors in different groups. Group I Acetone, Group II acetone+DMBA, Group III resveratrol (25 u03bcM)+DMBA, Group IV resveratrol (50 u03bcM)+DMBA, Group V acetone+ resveratrol (50 u03bcM). Equal loading was confirmed by reprobing the membrane with u03b2-actin ![file] Please compare with the control bands from Figures 1 and 2 of [Arora et al (2006)] . ![] , 
  Fig. 5. Western blots showing the effect of **resveratrol **on ... b **AKT **... in mouse skin tumors in different groups It is surprisingly easy to obtain 5(b) from Fig 2(b) of  Regulation of p21 ras protein expression by diallyl sulfide in DMBA induced neoplastic changes in mouse skin [(Arora et al 2006)] . ![file] There is the minor complication that 2(b) represents  **p21 ras** expression... in **membrane fractions** , and recycles a different loading control. , 
 #7 The authorship also overlaps with the list involved in a February retraction of a 2007 paper, where the main reason was the same: re-use of the same limited repertoire of loading controls.  [Retraction notice to  Theaflavins induced apoptosis of LNCaP cells is mediated through induction of p53, down-regulation of NF-kappa B and mitogen-activated protein kinases pathways]  [Life Sci. 18 23 (2007) 2137-2146] This article has been retracted at the request of the Editor-in-Chief. Multiple figures in this article appear to be falsified fabricated. Figure 2A. The images appear to be rotated and reused. Figure 3A. The Beta-actin band appears to be identical to articles from multiple other journals. (doi:10.1016 j.canlet.2005.10.049; doi:10.1002 mnfr.20070019; doi:10.1007 s11095-008-9723-z; doi:10.1016 j.lfs.2007.04.009; doi:10.1080 01635580701613772) Figure 6. The Beta-actin band appears to have been reused and possibly manipulated. Relevant pubpeer thread here: https: pubpeer.com publications 0D63933A222787F47952520C798C52 , 
 #6 On that same theme, it is remarkably easy to obtain a blot for Cytochrome C (present figure Fig 3(d), below) from one for  Apaf-1 : Figure 5(B) from  Resveratrol induces apoptosis involving mitochondrial pathways in mouse skin tumorigenesis [(Kalra et al 2008)] . ![] And who wants a contrast-enhance version of Fig 3(a)? ![file] , 
 No doubt most readers are aware of this already, but if not: https: journals.plos.org plosone article?id=10.1371 journal.pone.0215980 (Retraction of article with some shared authors). Notice reads (emphasis added): u201cRetraction: Resveratrol and Black Tea Polyphenol Combination Synergistically Suppress Mouse Skin Tumors Growth by Inhibition of Activated MAPKs and p53 The PLOS ONE Staff Following the publication of this article [1], the following concerns were raised: Figure 3A panel IV and Figure 3B panel IV appear to contain areas of similarity; Figure 3A panels I, V, and VI appear to contain areas of similarity; Figure 3B, panels I and II contain areas of similarity; Figure 3B, parts of panels V and VI contain areas of similarity; Figure 6 of a previously published article in the Journal of Food and Chemical Toxicology [2] by four of the authors and Figure 6 in this article [1] appear to contain similar figures: Figure 6a in [2] appears similar to Figure 6(I) in [1]; Figure 6c in [2] appears similar to Figure 6(III) in [1]; Repeated regions of similarity in flow cytometry plots within Fig 6 IV, VI, and II; Figure 2A ERK1 2 (Total) bands in lanes 5 and 6 appear similar when adjusted for brightness contrast; Figure 2A p38 (Total) panel lanes 2 and 6 appear similar when adjusted for brightness contrast; Figure 2A ERK1 (P) panel contains vertical discontinuities across all lanes when adjusted for brightness contrast; Figure 5 p53 contains background irregularities when adjusted for brightness contrast such that lane 2 appears to have a background different to other lanes. **The authors have been unable to provide any primary data underlying the figures.** The authors commented that the regions of similarity in histopathological images arose due to similarity in lesions studied. For the similarities noted in flow cytometry plots, the authors commented that this could have arisen due to similarity in equipment and protocols. In the absence of the data underlying the figures and in light of the above concerns, the PLOS ONE Editors retract the article.u201d The authors did not comment on the retraction decision. Citation: The PLOS ONE Staff (2019) Retraction: Resveratrol and Black Tea Polyphenol Combination Synergistically Suppress Mouse Skin Tumors Growth by Inhibition of Activated MAPKs and p53. PLoS ONE 14(4): e0215980. doi:10.1371 journal.pone.0215980 Published: April 19, 2019 , 
 2021 retraction. https: link.springer.com article 10.1007 s11095-020-02973-y Retraction Note: Pharmaceutical Research volume 26, pages 211u2013217(2009) https: doi.org 10.1007 s11095-008-9723 The Editor-in-Chief has retracted this article [1] due to concerns about the reliability of the figures in this article. Specifically: For Fig. 3A, the beta-actin band appears to be identical to figures from multiple other journals [2, 3, 4, 5, 6] For Fig. 3C, an identical beta-actin band appears in Fig. 1B of [2] despite different conditions For Fig. 3D, The first two and last two lanes of the beta-actin band appear to be identical For Fig. 3D, the Cytochrome C blot appears to have be a duplication of the Apaf 1 band in Fig. 5B from [7]. Lanes III and IV seem to have been rotated and vertically stretched. For Fig. 4C, the beta-actin band appears remarkably similar to Figs. 1 and 2 in [3] For Fig. 5B, lanes I to V of the AKT blot appear to have been duplicated from lanes A to E in Fig. 2B of [3]. Lanes III and IV seem to have been rotated and vertically stretched. Given these concerns, the Editor-in-Chief therefore no longer has confidence in the overall study presented in this article. Authors Jasmine George and Yogeshwer Shukla do not agree with this retraction. Preeti Roy, Neetu Kalra, and Sahdeo Prasad have not responded to notices about this retraction. 1. Roy, P., Kalra, N., Prasad, S. et al. Chemopreventive Potential of Resveratrol in Mouse Skin Tumors Through Regulation of Mitochondrial and PI3K AKT Signaling Pathways. Pharm Res 26, 211u2013217 (2009). https: doi.org 10.1007 s11095-008-9723-z 2. Arora, A., Siddiqui, I. A., Shukla, Y. (2004). Modulation of p53 in 7, 12-dimethylbenz [a] anthraceneu2013induced skin tumors by diallyl sulfide in Swiss albino mice. Molecular Cancer Therapeutics, 3(11), 1459u20131466. 3. Arora, A., Kalra, N., Shukla, Y. (2006). Regulation of p21 ras protein expression by diallyl sulfide in DMBA induced neoplastic changes in mouse skin. Cancer letters, 242(1), 28u201336. 4. Shukla, Y., Prasad, S., Tripathi, C., Singh, M., George, J., Kalra, N. (2007). In vitro and in vivo modulation of testosterone mediated alterations in apoptosis related proteins by [6]-gingerol. Molecular nutrition food research, 51(12), 1492u20131502. 5. Kalra, N., Seth, K., Prasad, S., Singh, M., Pant, A. B., Shukla, Y. (2007). Theaflavins induced apoptosis of LNCaP cells is mediated through induction of p53, down-regulation of NF-kappa B and mitogen-activated protein kinases pathways. Life sciences, 80(23), 2137u20132146. 6. Prasad, S., Kalra, N., Shukla, Y. (2007). Induction of apoptosis by lupeol and mango extract in mouse prostate and LNCaP cells. Nutrition and cancer, 60(1), 120u2013130. 7. Kalra, N., Roy, P., Prasad, S., Shukla, Y. (2008). Resveratrol induces apoptosis involving mitochondrial pathways in mouse skin tumorigenesis. Life sciences, 82(7u20138), 348u2013358. ,