Retracted: MicroRNA‐300 promotes apoptosis and inhibits proliferation, migration, invasion and epithelial‐mesenchymal transition via the Wnt/β‐catenin signaling pathway by targeting CUL4B in pancreatic cancer cells
Authors: Yi Zhu
Xia-Xing Deng
Jiang-Ning Gu
Bai-Yong Shen
Hao Chen
Dong-Feng Cheng
Shi Chen
Jia-Qiang Zhang
Qian Zhan
Cheng-Hong Peng
Journal: Journal of Cellular Biochemistry (2018)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: CUL4B; Wnt/β-catenin signaling pathway; epithelial-mesenchymal transition; invasion; microRNA-300
Affiliations: Department of General Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, P.R. China., Research Institute of Digestive Surgery, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, P.R. China., Department of Hepatobiliary Surgery, Fujian Provincial Hospital, Fujian Medical University, Fuzhou, P.R. China., Department of General Surgery, the Second Hospital of Zhejiang University, Hangzhou, P.R. China.

Abstract:  The study aims to verify the hypothesis that up-regulation of microRNA-300 (miR-300) targeting CUL4B promotes apoptosis and suppresses proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of pancreatic cancer cells by regulating the Wnt/β-catenin signaling pathway. Pancreatic cancer tissues and adjacent tissues were collected from 110 pancreatic cancer patients. Expression of miR-300, CUL4B, Wnt, β-catenin, E-cadherin, N-cadherin, Snail, GSK-3β, and CyclinD1 were detected using qRT-PCR and Western blot. CFPAC-1, Capan-1, and PANC-1 were classified into blank, negative control (NC), miR-300 mimics, miR-300 inhibitors, siRNA-CUL4B, and miR-300 inhibitors + siRNA-CUL4B groups. The proliferation, migration, invasion abilities, the cell cycle distribution, and apoptosis rates were measured in CCK-8 and Transwell assays. Pancreatic cancer tissues showed increased CUL4B expression but decreased miR-300 expression. When miR-300 was lowly expressed, CUL4B was upregulated which in-turn activated the Wnt/β-catenin pathway to protect the β-catenin expression and thus induce EMT. When miR-300 was highly expressed, CUL4B was downregulated which in-turn inhibited the Wnt/β-catenin pathway to prevent EMT. Weakened cell migration and invasion abilities and enhanced apoptosis were observed in the CUL4B group. The miR-300 inhibitors group exhibited an evident increase in growth rate accompanied the largest tumor volume. Smaller tumor volume and slower growth rate were observed in the miR-300 mimics and siRNA-CUL4B group. Our study concludes that lowly expressed miR-300 may contribute to highly expressed CUL4B activating the Wnt/β-catenin signaling pathway and further stimulating EMT, thus promoting proliferation and migration but suppressing apoptosis of pancreatic cancer cells. 

Retracted: Yes

PubPeer comments:
 Just an example of a reused and cut Western. Probably there are more examples in this paper. Just look at band shapes and background: ![file] , 
 #1 The protein blobs in that figure seem to come from the same restricted gamut shown in this montage: ![] Fig 6B,  protein band graph , from  MicroRNA-374 Exerts Protective Effects by Inhibiting SP1 Through Activating the PI3K Akt Pathway in Rat Models of Myocardial Ischemia-Reperfusion After Sevoflurane Preconditioning [(Zhang et al 2018)] . Fig 4D,  GLP-1R and CREB proteins in the myocardium of mice in each group examined by western blotting , from  Suppression of Long Non-Coding RNA LINC00652 Restores Sevoflurane-Induced Cardioprotection Against Myocardial Ischemia-Reperfusion Injury by Targeting GLP-1R Through the cAMP PKA Pathway in Mice [(Zhang et al 2018)] . Fig. 6B,  ELISA and western blot analysis were applied to determine the miR- 495 expression and mRNA and protein expression of NLRP3, TNF-u03b1, IL-1u03b2, IL-18 and caspase-1 , from  MicroRNA-495 Ameliorates Cardiac Microvascular Endothelial Cell Injury and Inflammatory Reaction by Suppressing the NLRP3 Inflammasome Signaling Pathway [(Zhou et al 2018)] . Fig 6B,  WB images showing protein expression of different genes in each group , from  MicroRNA-27a Suppresses Detrusor Fibrosis in Streptozotocin-Induced Diabetic Rats by Targeting PRKAA2 Through the TGF-u03b21 Smad3 Signaling Pathway . [(Wu et al 2018)] . Fig 11E,  protein bands of each factor in ASMCs of asthma mice in each group detected by western blot analysis , from  MicroRNA-485 Modulates the TGF-u03b2 Smads Signaling Pathway in Chronic Asthmatic Mice by Targeting Smurf2 . [(Wang et al 2018)] . Or Figs 5B and 10B of [Liu et al (2018)] : ![] , 
 Fig 8A:  Proliferation and invasion ability of cells in all groups after cell transfection detected by Transwell assay Notes: A, comparison of migration and invasion abilities of cells among all groups (u00d7 200) . Possible duplicate cells in one panel. ![file] There is also a peculiar coincidence in the Blank and NC panels. 5 cells are identically located (relative to one another) in both panels, so after color-reversing the NC panel and superimposing it on the Blank, they cancel out and disappear. Yet other cells, and the background panel of membrane pores, are different. ![file] , 
 #3 Perhaps more duplicates (see green)? *** Enhanced saturation etc ![file] , 
 Fig 9A.  Comparison of cell cycles and apoptosis rates among six groups after cell transfection Notes: A, cell cycle of cells in all groups when observed under a microscope *[sic]* ![file] Surprising similarities between some of these flow-cytometry histograms (or what is visible of them behind the modeled distributions) and those if Fig 9A,C of [Qian et al (2018)] . ![file] , 
 #4 Two panels in the figure also appear in Fig 12A of [Wu et al (2018)] ,  Comparison of invasive tumor cells in SCE Note: A, Microscopic observation of invasive cells . ![file] , 
 Dear Hoya, Thanks for your attention. We would like to respond to your comments one by one. Response to #2: We appreciate that you have brought this to our attention. It seems that the wrong blot was inadvertently included when we assembled the final figure. Weu2019ll examine our raw data to check it out. Since the experiments were performed four or five years ago, it will take some time to sort out the data. Once confirmed, we are ready to work with the journal to correct this figure. Response to #3: The distributions of individual cells are similar, but each cell takes diverse shapes. This phenomenon can t be ruled out as accidental. Weu2019ll examine our raw data to check it out. Also, we are delighted to reperform the experiments to further verify our results. Response to #5 6: We find that our article has been published online in July 2017, while the paper of Qian et al. was accepted by a different journal on 22 Mar 2018. Likewise, Wuu2019s paper was received by another journal on 23 Sep 2017 and accepted on 2 Dec 2017, showing that the related articles were published later than ours. Whatu2019s more, each transwell assay photo similar to our figures in Wuu2019s paper was actually a local image of ours. Based on these facts, it is quite possible that some of our images may be leaked for as-yet-unclear reasons. Thank you again for your valuable comments! , 
 #7 Regarding your statement that the cells are similar but not the same: could the authors post links here to higher resolution images of the original experiments (perhaps using https: imgur.com or something similar)? That should help reassure readers. , 
 Fig 6E again: here at right. At left, Fig 4B from [Wen et al (2018)] :  The interaction between SPRY4-IT1 expression and EZH2 expression detected by RT-qPCR and western blotting ... B and E, the SPRY4-IT1 protein bands in the eluant of RNA pull-down assay in A549 ... detected by western blotting; Positive control was the RNA provided by kit that could specially combine with HuR protein, and negative control was poly(A)25 RNA without any combination; anti-sense RNA was designed for SPRY4-IT1, which could compete with EZH2 to combine with SPRY4-IT1 . Loading control bands are more similar than I would have expected. ![file] Throwing Fig 5B from Wen et al into the mix as well *[now at left]* :  Expressions of miR-708, bone morphogenetic protein and activin membrane-bound inhibitor (BAMBI), Wnt10B, P53, Bcl-2, VEGF, Fas, Bax, caspase-3, and cleaved caspase-3 in each group. A, Histogram of mRNA expressions. B, Electrophoretograms of protein expressions. ![file] Update: The control bands of Fig 6A C of [Wang et al (2018)] also show some similarities. ![] , 
 Fig 8A again. ![] For comparison with the right-hand panel of Fig 10A of [Wang et al (2017)] .  The number of invasive cells and cell migration rate among the control, NC and Bmi-1 shRNA groups Notes: A, images of cell invasion in the control, NC and Bmi-1 shRNA groups . ![] One or two cells appear to have been added or removed. , 
 At right, Fig 1A,  Positive expression of CUL4B protein in pancreatic cancer tissues and adjacent tissues . At left, Fig 2A from [He et al (2018)] ,  positive expression of CUL4B and u03b2-catenin proteins detected by immunohistochemistry . ![file] This comment was brought to you by the Department of Unexpected Similarities. , 
 Retracted [30 June 2021] . --------------------------------- The above article, published online on 07 July 2017 in Wiley Online Library has been retracted by agreement between the the journal s Editor in Chief, Prof. Dr. Christian Behl, and Wiley Periodicals LLC. The retraction has been agreed following an investigation based on allegations raised by a third party. A detailed investigation revealed that several image elements of the experimental data were published elsewhere in a different scientific context. Thus, the editors consider the conclusions of this article to be invalid. ,