Analysis of apoptosis methods recently used in cancer research and cell death & disease publications

Analysis of apoptosis methods recently used in cancer research and cell death & disease publications

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Over several decades, significant advances have been made in implementing several morphological and biochemical criteria to define and characterize apoptosis. In order to appropriately


identify apoptosis in cellular cultures or _in vivo_ (animal models), with the ultimate aim of discovering novel, useful, specific, and powerful pro-apoptotic antitumoral drugs, it is


necessary to accurately validate apoptotic processes through morphological, biochemical, or immunological methods. Often, apoptosis is detected and/or measured by only one method and the


authors frequently use imprecise terminology, such as ‘% apoptosis’, ‘% cell death’, instead of defining the specific method used such as ‘% cells with condensed chromatin’, ‘% cells TUNEL


positive’, ‘% cells Annexin V positive’ and others.1 The Nomenclature Committee on Cell Death (NCCD) recommends and encourages researchers to demonstrate that apoptosis or other forms of


cell death take place using more than one assay, as an artifact removal feature, and to avoid the ‘confusing and imprecise’ nomenclature, such as ‘% apoptosis’, which ‘should definitively be


abandoned’.1 Moreover, the NCCD ‘urges’ all life science journals to join the Cell Death and Differentiation journal in adopting these recommendations.1 In order to determine the level of


compliance with these recommendations, we analyzed 110 _Cancer Research_ articles that detect/measure apoptosis, published between August 15, 2010 and February 15, 2011, and 120 similar


research articles published in _Cell Death & Disease_ between January 1, 2010 and September 2011. In 21 of the _Cancer Research_ and 15 of the _Cell Death & Disease_ articles,


apoptosis was determined by the authors for different treatments or in different settings using a different combination of apoptosis techniques each time, thus leading the total number of


apoptotic determinations (entries) to 137 (_Cancer Research_) and 136 (_Cell Death & Disease_) (see Table 1 and Table 2). The results clearly show that in only 60 of the 137 entries


(43.8%) for _Cancer Research_ and 96 of the 136 entries (70.58%) for _Cell Death & Disease_, the authors used at least two different methods for apoptosis detection. In addition, we


determined that in 5 of the 137 (3.64%) entries from _Cancer Research_ and in 34 of the 136 (25%) of the entries from _Cell Death & Disease_ the authors use at least three methods for


apoptosis detection (Figure 1). Moreover, it is important to emphasize that most, if not all of the apoptosis techniques used to detect apoptosis as single methods may, as shown by some


reports, also detect necrosis or other cellular processes. As an example, DNA fragmentation measured by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL)


(26 of 137 from _Cancer Research_; 26 of 136 from _Cell Death & Disease_) or subG1 DNA content (30 of 137 from _Cancer Research_; 18 of 136 from _Cell Death & Disease_) or Annexin V


alone, without PI/7AAD (16 of 137 from _Cancer Research_; 7 of 136 from _Cell Death & Disease)_ and other assays can also detect necrosis.1, 2, 3, 4, 5, 6 In addition, caspases can be


activated during cellular processes other than apoptosis.1 Thus, more than one method for apoptosis detection should be used. In order to investigate the proper use of the specific name for


each method used, we analyzed the same entries described above. Our analysis shows similar results for the two journals. While for _Cell Death & Disease_, 83 from 136 entries (61.02%)


use the name of the specific method used for apoptosis detection, among the 137 entries from _Cancer Research_, 87 (63.5%) use the name of the specific method used (Figure 1). These results


certainly leave space for improvement. In conclusion, these results clearly suggest the need for improvements and of adequate guidelines for authors, reviewers, and editors regarding the


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Article  CAS  PubMed Central  PubMed  Google Scholar  Download references ACKNOWLEDGEMENTS OB received a fellowship from the Lady TATA Memorial Trust, London, UK. This work was also


supported by the National Institutes of Health grants CA127264 (to AA), CA105306, CA131664, and HL080192 (to RK). AUTHOR INFORMATION Author notes * O Bucur and A L Stancu: These authors


contributed equally to this work. AUTHORS AND AFFILIATIONS * Department of Pathology, Harvard Medical School and Beth Israel Deaconess Medical Center, Boston, MA, USA O Bucur, A L Stancu 


& R Khosravi-Far * Department of Molecular and Cell Biology, Institute of Biochemistry of the Romanian Academy, Bucharest, Romania O Bucur * Carol Davila University of Medicine and


Pharmacy, Bucharest, Romania A L Stancu * Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USA A Almasan Authors * O Bucur View author


publications You can also search for this author inPubMed Google Scholar * A L Stancu View author publications You can also search for this author inPubMed Google Scholar * R Khosravi-Far


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CORRESPONDING AUTHORS Correspondence to R Khosravi-Far or A Almasan. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no conflict of interest. RIGHTS AND PERMISSIONS This work is


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