A novel human phosphatidylethanolamine-binding protein resists tumor necrosis factor alpha-induced apoptosis by inhibiting mitogen-activated protein kinase pathway activation and phosphatidylethanolamine externalization
Authors: Jianming Qiu
Hongying Sun
Bin Liu
Xiaojian Wang
Taoyong Chen
Lihuang Zhang
Hongzhe Li
Nan Li
Tao Wan
Xuetao Cao
Journal: Journal of Biological Chemistry (2004)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: phosphatidylethanolamine-binding proteins; hPEBP4; tumor cells; TNFalpha; Raf-1
Affiliations: Institute of Immunology, Zhejiang University, Hangzhou 310031, People's Republic of China.

Abstract:  The phosphatidylethanolamine (PE)-binding proteins (PEBPs) are an evolutionarily conserved family of proteins with pivotal biological functions. Here we describe the cloning and functional characterization of a novel family member, human phosphatidylethanolamine-binding protein 4 (hPEBP4). hPEBP4 is expressed in most human tissues and highly expressed in tumor cells. Its expression in tumor cells is further enhanced upon tumor necrosis factor (TNF) alpha treatment, whereas hPEBP4 normally co-localizes with lysosomes, TNFalpha stimulation triggers its transfer to the cell membrane, where it binds to Raf-1 and MEK1. L929 cells overexpressing hPEBP4 are resistant to both TNFalpha-induced ERK1/2, MEK1, and JNK activation and TNFalpha-mediated apoptosis. Co-precipitation and in vitro protein binding assay demonstrated that hPEBP4 interacts with Raf-1 and MEK1. A truncated form of hPEBP4, lacking the PE-binding domain, maintains lysosomal co-localization but has no effect on cellular responses to TNFalpha. Given that MCF-7 breast cancer cells expressed hPEBP4 at a high level, small interfering RNA was used to silence the expression of hPEBP4. We demonstrated that down-regulation of hPEBP4 expression sensitizes MCF-7 breast cancer cells to TNFalpha-induced apoptosis. hPEBP4 appears to promote cellular resistance to TNF-induced apoptosis by inhibiting activation of the Raf-1/MEK/ERK pathway, JNK, and PE externalization, and the conserved region of PE-binding domain appears to play a vital role in this biological activity of hPEBP4. 

Retracted: Yes

PubPeer comments:
 Several issues with this paper were reported to the journal in April 2014, more than 5 years ago. * Figure 5. Two flow cytometry panels presented in Figure 5A look very similar to each other. ![file] , 
 * Figure 7B. Several groups of dots appear to be visible multiple times within the same panel. ![file] , 
 Figure 7B. * Top left panel: Several groups of dots appear to be visible multiple time within this panel. ![file] * Center two panels: These two panels look unexpectedly similar to each other, while some areas appear to be visible multiple times within the same panel. ![file] * Top right and bottom left panel: These also look unexpectedly similar to each other, while some areas appear to be visible multiple times within the same panel. ![file] , 
 #1 Dr. Bik, Might there be more to this paper than #FlawCytometry? For example, 5 years ago, did you also report that some of the gel bands in Figure 6B look rather similar to one another? And can you also, as I can, visualise the gel splicing artefacts? ![image] , 
 Dear Condylocarpon, I am not sure why you ask me these questions. They might be better directed to the authors of this paper. , 
 #4 The half-tone nature and low resolution of Fig 6B are regrettable, and rule out any kind of close comparison. But after 15 years we can t really demand that the authors provide the original data. ![file] , 
 #6 It is also fair to notice the similarity between the actin band in Fig 6B and that of the original version of Fig 3B in [Wang et al (2005)] . ![file] , 
 #5 Thank you for your rapid reply. (Feel free to take your time in future - I do.) I have no plans to contact the authors: I think, by now, they know where to look. If memory serves, five years ago when you first contacted the journal, JBC had no Data Integrity Manager (aka DIM)? However, their response to the flood of image duplications posted to teh Intarwebs was that they eventually got around to hiring said DIM. And ever since then, they have been a poster child for how a journal might seek to improve the scientific record. Not many journals have retracted 9 articles in one day for image fraud, all coming from one of the worldu2019s toppest of top institutes. JBC did and I love them to death. https: retractionwatch.com 2017 05 25 journal-retracts-nine-papers-one-day-author-investigation-weizmann-institute The DIMu2019s eyes are surely just as sharp as yours. But she has a lot on her plate and her responsibilities are greater than ours as she dare not make a serious mistake. She can surely go faster if the donkey work to identify issues in the set of JBC papers from this particular cohort can be done by the community. Thus, my preference, as regards JBC specifically, is that we (the community) work through the figures of each paper of interest before emailing in our demands for the paper to be reviewed. I believe that the more information that JBC has, the faster that we will see results. By results, I mean improvements to the scientific record. I do not think it is desirable to present the JBC DIM with a continually moving target, each time a new issue is spotted. Iu2019m slow and Iu2019m easily distracted from improving on scientific knowledge by the mundane demands of the day job. But even I can see that there is more to report in this JBC paper ![image] , 
 #6 I think you are being too kind to Fig. 6B, Hoya. In the image posted in #4, seek out the hungry primeval swampfish in hPEBP4 and see how it has been separated from its prey band by a pallid vertical line and then compare to the distance of the predator prey in the mock equivalents to right. But wait, thereu2019s more. So let us move on to Fig. 2A, B, C. By the time that these Shanghaied RT-PCR gels have been deconstructed, it should be clear that they have almost as many replicants as does the Terracotta Army. For example, the cunningly disguised beta-Actin replicants in Fig. 2A and Fig. 2C. But letu2019s not get ahead of ourselves since, ideally, one should begin at the beginning and indeed Fig. 2A really does need to be marked up all by itself ![image] , 
 #9 Supplementary Figure S2B from [Wang et al (2008)] : ![] , 
 #10 Marvellous! Take an existing gel image and elongate it with its own already duplicated bands. Luckily the 2008 paper is already retracted so that JBC does not have a copyright issue with JBC. Here are the lane duplications in the Shanghaied RT-PCR gels of Fig. 2B ![image] , 
 As might be expected by now, a Shanghaied ImmunoBlot can also be espied in Fig. 6A. The SIB is presented after applying the LUT of 6 Shades to the image using FIJI ![image] , 
 #12 RE: *  Shanghaied RT-PCR gels *  Shanghaied ImmunoBlot *  the Terracotta Army Please reconsider using elements of nationality, ethnicity, culture, etc., as the basis for derogatory remarks. Some may justifiably find it offensive. The scientific critiques are sufficiently convincing without these embellishments. Thank you. , 
 #13 *Leucanella*, I very much value your postings at PubPeer. You have excellent image analysis skills, certainly superior to mine. Not only have you often commented upon obvious image problems but you also regularly correct commenters when their suggestions are completely wrong, or are doubtful, or are simply far too weak to post. Although my remarks are not always polite, they are posted so in defence of the scientific literature and they do tend to be accompanied by evidence of problems that I have identified. The issue I seek to address is that we have had all these decades of politely pointing out problem papers to journals, research institutes, funders, aka the scientific establishment. It did not work. On the other hand, exposure on the internet is now working. Slowly, but surely. I do feel we should not overdo the ad hominems and not presuppose in our comments about who did what in a problem paper with multiple authors. But if a paper has absurd figures, we should be able to say so and, in my opinion, we **should** say so. To do otherwise is a disservice to the scientific record that real scientists cherish and that they seek to contribute to. Confusingly for an obscure South American woody plant, *C. amazonicum* might or might not be British. Representative comments here about the UCL (aka Global Research Powerhouse) paper by u201cThe Masteru201d (which is, incidentally, the correct mode of address and therefore cannot be an ad hominem) https: pubpeer.com publications 68B2D4A92E2BC3BAF0BCB5E1C599AB Representative comments here about the appalling behaviour of the puffed up Lancet journal (and UCL again) regarding the **Warlocks of Degeneration** https: pubpeer.com publications 2968ADAFF1170575BBB3D0B74FD6CD Now, if Brexit happens, UCL wonu2019t be the self-hyped **Global Research Powerhouse**, they will instantly become the **Global Research Minnow**. UCLu2019s pretentious moniker will be taken down by the total collapse of the UKu2019s economy. Alas **Global Research Powerhouse** we hardly knew ye. Moving out of the UK, I fail to see how any real scientist could be polite about the Weizmannu2019s Extra-Cellular Radio-Controlled Kinase mobile phone scaremongering paper that they do nothing about https: pubpeer.com publications 86E709835E96908738330ED4B849CC And then there is the the Weizmannu2019s elephant in the room paper which launched a worldwide abuse of laboratory mice in the failed search of mouse-human early embryonic chimeras that canu2019t happen. https: pubpeer.com publications F9DC43608AA039D4F514706B2188BA Coming back to China, it urgently needs to reform its publication incentives. Paying researchers because they have published in CNS journals is as bonkers as it gets. *All metrics of scientific evaluation are bound to be abused. Goodhart s law (named after the British economist who may have been the first to announce it) states that when a feature of the economy is picked as an indicator of the economy, then it inexorably ceases to function as that indicator because people start to game it.* https: en.wikipedia.org wiki Goodhart%27s_law China knows that it is seeing many hundreds of retractions from the scientific literature. Image fabrications, fake review circles and so forth. But does it know why? Strange but true: More actual quality requires less publications. At least in the short term, *Leucanella*, your wish may be met: In this Brexit-benighted moment of time, I will not be commenting here much. Which I regret, since the opportunity to work on improving the scientific record is with us, right here and now. Anyway, regarding this Shanghaied JBC paper, most has been discovered and it is therefore time to contact the DIM with our findings. The Fig. 2C control has a tell-tale half-moon splice edge shared with Fig. 2A and the already retracted 2008 s2B image posted by Hoya (#10). ![image] , 
 September 2020: This article has been withdrawn. https: www.jbc.org content 295 39 13692  This article has been withdrawn by the authors except for Hongzhe Li, who could not be reached. In Fig. 2A, lanes 3 and 4 of the hPEBP4 panel and lanes 3, 4, and 6 of the actin panel are the same. In the hPEBP4 panel of Fig. 2B, the last two lanes are the same. In the actin panel for the same figure, lanes 3 and 4 are duplicates and lanes 6 and 7 are duplicates. In Fig. 2C, lanes 3 and 4 of the actin panel are duplicates. Lanes 8 and 9 of the same panel are duplicates. Lanes 1 and 3 of the myc immunoblot in Fig. 3C are the same. The last lane of the actin immunoblot in Fig. 4A was reused in the first lane of the actin immunoblot in Fig. 4B. The first and last lanes of the Raf-1 immunoblot in Fig. 4C are the same. A portion of the FACS plot shown in the top middle panel of Fig. 5A was reused in the bottom left panel. The lower phospho-JNK band in lanes 5 and 12 in Fig. 6A are the same. Lanes 6 and 7 of the actin immunoblot were reused in lanes 10 and 11. The withdrawing authors do not agree with some of the Journal s analyses and state that their analysis shows differences in some of the concerned bands that exclude reuse of the same bands. The withdrawing authors state that they repeated all of the relevant experiments and that the results are the same in their hands. The withdrawing authors state that the issues described above do not affect the interpretation of the data or conclusions of this work. This article describes the novel molecule hPEBP4 and characterizes its apoptosis-resistant function in breast cancer. The withdrawing authors state that its function and mechanism have been confirmed in breast cancer and other types of cancer by other independent studies (Garcia, R., et al. (2009) EMBO Rep. 10, 278u2013284; Wang, S., et al. (2017) Biomed. Pharmacother. 90, 659u2013664; Huang, R., et al. (2019) Mol. Carcinog. 58, 135u2013143; and Yu, G., et al. (2013) Tumor Biol. 34, 91u201398). The withdrawing authors thus stand by the overall conclusion of the study. , 
 https: retractionwatch.com 2020 09 30 author-says-misguided-efforts-for-the-ideal-western-blot-led-to-the-withdrawal-of-these-studies ,