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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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http://creativecommons.org/licenses/by-nc-nd/3.0/ Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Bucur, O., Stancu, A., Khosravi-Far, R. _et al._ Analysis of apoptosis methods
recently used in _Cancer Research_ and _Cell Death & Disease_ publications. _Cell Death Dis_ 3, e263 (2012). https://doi.org/10.1038/cddis.2012.2 Download citation * Published: 02
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