Retraction Watch

Tracking retractions as a window into the scientific process

Bogus Western blots lead to retraction of cancer paper

with 22 comments

ejpharmA group of Italian researchers has retracted their 2013 paper on colorectal cancer because one of the authors, they, say, was tinkering with the data.

The article, “PBOX-15 induces apoptosis and improves the efficacy of oxaliplatin in human colorectal cancer cell lines,” appeared in the European Journal of Pharmacology in August. The first author was Giuseppina Gangemi, of the University of Salerno.

The paper purported to find that:

An emerging new class of targeted therapeutic molecules against the enzyme fatty acid amide hydrolase (FAAH) is a novel series of pyrrolo-1,5-benzoxa(thia)zepine compounds. A member of this family, pyrrolo-1,5-benzoxazepine-15 (PBOX-15), is a tubulin depolymerizing agent displaying a proapoptotic activity in a variety of human tumor cell types, including those derived from both solid and hematological malignancies, with minimal toxicity towards normal blood and bone marrow cells. In this study, we evaluated the PBOX-15-mediated effects in human colorectal cancer cell (CRC) lines. The compound, used at doses equal to or greater than 1 μM inhibits the proliferation of human CRC cell lines in a dose- and time-dependent manner, inducing a significant cell cycle arrest in the G2/M phase. DNA fragmentation assays and western blot analysis demonstrated that treatments prolonged over 48 h triggered a strong activation of the intrinsic apoptotic pathway as indicated by activation of caspase-3, caspase-9 and PARP. Moreover, nanomolar doses of PBOX-15, unable to cause microtubule depolymerization, significantly improved the oxaliplatin and 5-fluouracil-induced anti-proliferative effects in CRC cell lines. These results showed, for the first time, that PBOX-15 represents a promising compound for the treatment of human CRC and a strong candidate for novel therapeutic options.

Alas, PBOX-15’s glory must be deferred. According to the retraction notice:

Upon close inspection, the co-authors realized that the first author (Giuseppina Gangemi) substantially manipulated some of the western blot figures reported in the paper. None of the other authors are responsible for this occurrence. Even though the main conclusions are not affected by such manipulations, they wish to retract this publication, and apologize for the confusion and inconvenience caused.

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  • ferniglab March 28, 2014 at 11:44 am

    Rather opaque notice – has anyone looked at the paper to see what evidence there is? Pubpeer only has the retraction notice

  • Seshat March 28, 2014 at 1:48 pm

    Here is another one: the Cleaved Caspase-3 panels shown in Figure 5A and 5B appear to be from the same gel. Different exposure times, but an air bubble in the 3rd band and a smudge in the 8th band are identical.

  • Seshat March 28, 2014 at 2:07 pm

    But wait, there’s more.

    The beta actin panels in Figure 5A and 5B also appear to be identical, but 180 degrees rotated.

    In Figure 6, the top (DLD-1) and bottom (HT29) ERK blot panels might be the same, but 180 degrees rotated. It’s hard to see with that big orange “RETRACTED” watermark, but there is a small smudge in the first lane in the top ERK panel that is also seen in the left lane in the bottom ERK panel.

  • Dan Zabetakis March 28, 2014 at 5:06 pm

    As has been pointed out above, there are several major issues with the data in this paper. I have a couple of questions.

    1) There are many authors on this paper. How is it that none of them noticed the problems prior to submission?

    2) The retraction states that the first author was responsible. Does the first author agree with that? Does no one else really share any responsibility?

    3) The retraction states that the main conclusions are not effected. How do they know that? The data in this paper seems to have been so thoroughly tainted that the remaining authors cannot be sure what the actual results would have been.

    • david hardman March 29, 2014 at 3:49 am

      Mol Pharmacol. 2008 Dec;74(6):1678-86. doi: 10.1124/mol.108.049205. Epub 2008 Sep 18.
      The cannabinoid CB1 receptor antagonist rimonabant stimulates 2-deoxyglucose uptake in skeletal muscle cells by regulating the expression of phosphatidylinositol-3-kinase.
      Esposito I, Proto MC, Gazzerro P, Laezza C, Miele C, Alberobello AT, D’Esposito V, Beguinot F, Formisano P, Bifulco M.

      Dipartimento di Biologia e Patologia Cellulare e Molecolare & Istituto di Endocrinologia ed Oncologia Sperimentale del Consiglio Nazionale delle Ricerche, Università degli Studi di Napoli Federico II, Napoli, Italy.

      Figure 1.

      Figure 1B. WB.: CB1. Vertical, straight changes in background between lanes 2,3 and 4.

      W.B.: beta-actin. No vertical changes in background.

      Figure 3.

      W.B.:p110. Vertical, straight change background between lower halves left and right lanes.

      Lower W.B.:p85(5 lanes). Please compare bands lanes 1 and 2.

      Figure 5.

      Figure 5A. W.B.:beta-actin. Please compare signal lanes 2 and 3.

      Figure 7.

      W.B.:p-Akt. Please compare bands lanes 2 and 4.

      W.B.:Akt. Please compare bands lanes 1,3 and 4.

      Figure 8.

      Right W.B.:PKCzeta panel. Please compare signal lanes 2 and 3.

      • david hardman March 29, 2014 at 3:51 am

        Mol Pharmacol. 2008 Dec;74(6):1678-86 at Pubpeer:

      • Seshat March 30, 2014 at 5:11 am

        Nice ones. Here are some more:
        Figure 3: compare b-actin lanes 1 and 6.
        Figure 7: compare PDK lanes 1/2 with 5/6.
        Figure 1: compare CB1 lanes 3,4 to Figure 5A b actin lanes 1,2 (mirror image and reshaped).

    • Jerry Lofti March 29, 2014 at 7:01 am

      “How is it that none of them noticed the problems prior to submission”
      Very good point. If you have a normal visual system, you notice immediately, at least the most obvious duplicates.

  • david hardman March 29, 2014 at 5:55 am

    Eur J Endocrinol. 2001 Nov;145(5):645-50.
    Lovastatin-induced apoptosis in thyroid cells: involvement of cytochrome c and lamin B.
    Di Matola T, D’Ascoli F, Luongo C, Bifulco M, Rossi G, Fenzi G, Vitale M.
    Author information

    Dipartimento di Biologia e Patologia Cellulare e Molecolare, Universita Federico II, Via S Pansini 5, 80131, Naples, Italy.

    Figure 3B. Please compare the backgrounds in the 12 and 24 hour lanes.

  • PPV March 31, 2014 at 11:57 am

    The FACS panels in the PBOX paper are also reused.

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