Journal of Cancer Science & Treatment

JCST, 1(1): 17-24

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Original Research Article: Open Access

Morphologic and Biochemical Criteria of Cell Death: Apoptosis and Necrosis

Michel Goldberg*
*
INSERM UMR-S 1124, Faculté des Sciences Fondamentaleset Biomédicales, Cellules souches, signalizationet prions, France.

Received March 01, 2018; Accepted April 16, 2018; Published January 28, 2019

ABSTRACT
Two hundred billion cells are dying every day. They need to be renewed. Dying or dead cell are involved a series of
apoptotic events or eventually displays necrosis. Four basic modes of cell death have been identified. Apoptosis purges no-
longer useful cells via phagocytosis, eliminating old cells, and/or stimulate tissue regeneration. Apoptotic cells are reflecting
a suicidal event, whereas necrosis is playing role as a homicide. Active programed processes are characterized as being an
autonomous happening. The process concerns caspases, which are inhibitors of apoptosis. According to morphologic and
biochemical criteria, caspases contribute to membrane blebbing, intra-cytoplasmic alterations and formation of cysteine-rich
domains. They are implicated in the translocation of nuclear proteins, play role in the control of the calcium efflux, regulate
membrane permeability and inhibit the translocation of mitochondrial membranes associated to apoptosis. DNA damages,
nuclear fragmentation leading to the formation of apoptotic bodies, autophagic cell death, necrosis and other forms of cellular
death are instrumental in the reliability of apoptotic events leading to cells death. Apoptosis appears to be crucial in
morphogenesis, and actually guide a series of measurable biochemical features involved in tissue renewal and regeneration.

INTRODUCTION
cells death types and their variant-derived are involved in
Apoptosis
purging no-longer useful cells from the host tissue or organ.
In 2009, the Nomenclature Committee on cell death These processes are followed by regeneration, wound
proposed a set of recommendations for the definition of healing, and also by scar formation. Biochemical methods
distinct cell death morphologies including ‘apoptosis’, have many advantages over morphological techniques
‘necrosis’ and mitotic catastrophe [1]. A functional (Figure 3).
classification of cell death sub routes includes extrinsic
Apoptosis purge no-longer useful cells from a tissue via
apoptosis, caspase-dependent or -independent intrinsic
phagocytosis, using scavengers and including macrophages.
apoptosis, regulated necrosis, autophagic cell death and
A crude estimation suggests that in the human body 60
mitotic catastrophe. Functional classification of cell death
billion cells die every day. The human body needs to yield
modalities have established links with anoikis, autophagic
60-86.4 billion new cells per day, compensating the cell loss.
cell death, caspase-dependent intrinsic apoptosis, caspase-
Cell renewal implicates that ∼200 billion of cells/per day die
independent intrinsic apoptosis, cornification, entosis,
and therefore have to be renewed. Signals from apoptotic
extrinsic apoptosis by death receptors, mitotic catastrophe,
cells to scavengers indicates that apoptosis is a suicidal
necroptosis, netosis and pyroptosis [1] (Figures 1-3).
event, whereas necrosis is an homicide.
Apoptosis and necrosis comprise different modes of cell Corresponding author: Michel Goldberg, INSERM UMR-S 1124, Faculté
death, respectively implicated in cell withdrawal and des Sciences Fondamentaleset Biomédicales, Cellules souches,
recovery. Different types of cell death have been described signalizationet prions. University Paris Descartes, Sorbonne, Paris Cité,
according to morphologic and biochemical criteria [2]. Four 75006 Paris France, Tel: 33 1 42 86 38 51, 33 1 6 62 67 67 09; E-mail:
michel.goldberg@parisdescartes.fr; mgoldod@gmail.com
basic modes of cell death have been identified following the
biomedical literature, together with ad-hoc variants adapted Citation: Goldberg M. (2019) Morphologic and Biochemical Criteria of
to different situations [3]. Cell Death: Apoptosis and Necrosis. J Cancer Sci Treatment, 1(1): 17-24.
Copyright: ©2019 Goldberg M. This is an open-access article distributed
These various situations include 1) apoptosis and 2) under the terms of the Creative Commons Attribution License, which
senescent death (SD). 3) Necrosis and 4) stress-induced cell permits unrestricted use, distribution, and reproduction in any medium,
death (SICD), that are actual procedures identifying cell provided the original author and source are credited.
death. Scavengers engulf SICD and necrosis. Four basic

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Figure 1. Pathways leading to cell death.

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Figure 2. Apoptosis versus Necrosis.

Figure 3. Mild to severe inflammation leads to pulp healing or to pulp regeneration.

Schematic representation of events contributing to necrosis, apoptosis and nemosis

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They are both irreconcilable to each other. In contrast with avoids inflammation. Necrosis has been characterized as a
apoptosis, necrosis eliminate plethoral cells and passive, accidental cell death resulting from environmental
consequently does not trigger regeneration [4]. perturbations with uncontrolled release of inflammatory
cellular contents. Other types of cellular death are
Cell death with some of the features of apoptosis may result
necroptose, self-digestion, mitotic catastrophe, anoikis,
from a variety of molecular pathways. Cell death follows
cannibalistic cell death, cornification or keratinization,
either apoptosis or necrosis. Apoptosis is described as an
pyroptosis, parthanatose and ferroptose (Figure 4).
active, programmed process of autonomous cellular that

Figure 4. Nemosis process leading to the formation of spheroid and the expression of cyclooxygenase-2, prostaglandin E2,
and interleukin-8.
Control of proliferation or cell cycle implicates caspases, of cellular volume (pyknosis), chromatin condensation,
Bcl-2, inhibitor of apoptosis (IAPs), FAK, membrane nuclear fragmentation (karyorrhexis), little or no
blebbing (ROCK1), gelsoline, lamines A, B, C, β-caténine, ultrastructural modification of cytoplasmic organelles,
ℽ-caténine, topoisomerase I and II. Extrinsic pathway of plasma membrane blebbing and engulfment by resident
apoptosis involves death ligands (Fas L, TNF, TWEAK, phagocytes (in vivo). It is worth noting that it is not correct
TRAIL), activation of death receptors (intracytoplasmic to assume that “programmed cell death” and “apoptosis” are
death domain and cysteine-rich domain) and intrinsic synonyms because cell death can manifest non-apoptotic
apoptosis (DNA damage, ROS, stress response), Anti- features.
apoptotic (BCL-2) anti- and pro-apoptotic events, pro-
Specific biochemical analyses (such as DNA ladders) should
apoptotic (Bax) seems to play role in apoptotic events. Anti-
not be employed as an exclusive means to define apoptosis,
apoptotic action of the Bcl-2 family inhibits the translocation
because this type of cell death may occur without
of some nuclear proteins in the nucleus, control the calcium
oligonucleosomal DNA fragmentation. The presence of
efflux from the rough endothelium, whereas antagonists of
caspases or the cleavage product of their substrates is not
ROS control the permeability or inhibit the translocation of
sufficient to define apoptosis. The measurement of DNA
mitochondrial membranes, which are also implicated in this
fragmentation and/or caspase activation helps to better
phenomenon.
define the type of cell death, through the ‘intrinsic’ or the
Apoptotic fragmentation ‘extrinsic’ pathway, with or without the contribution of
mitochondria.
Proposed by Kerr et al. [5], apoptosis was characterized by
nuclear and cytoplasmic condensation and cellular Cell death is frequently considered to be ‘caspase-
fragmentation into membrane-bound fragments (also named dependent’, suppressed by caspase inhibitors. Different
apoptotic bodies). Apoptosis is accompanied by the forms of apoptosis have been identified including apoptosis,
rounding-up of the cell, retraction of pseudopodes, reduction necroptosis, anoikis, pyronecrosis, death with autophagy or

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mitotic catastrophe, pyroptosis, cornification or Caspase-9, caspase-3 and caspase-7 have distinct roles
keratinization, entose (self-cannibalism), wallerian during intrinsic apoptosis [8]. Apoptosis is a form of
degeneracy, excitotoxicity, netose, parthanose and ferroptose programmed cell death that is regulated by the Bcl-2 family
[6]. and caspase family of proteins. Specific inhibition of
caspase-9 allows the efficient release of cytochrome c, but
Autophagy and autophagic cell death
blocks change in mitochondrial morphology and ROS
Degraded in phagosomes by caspases or cysteine-dependent production. Caspase-3-deficient MEFs are not resistant to
aspartate specific proteases. Interleukin-1β xov-1. Initiator intrinsic cell death. Altogether, the data suggest that caspase-
caspases (caspase-2, -8, -9 and -10) contain a small 9 is required for morphological change and ROS production
prodomain. Activated effector caspases selectively cleave a by cleaving and activating Bid into tBid. After activation by
restricted number of target proteins. One of the caspase-9, caspase-3 inhibits ROS production and is
morphological and biochemical associated with apoptosis, required for efficient execution of apoptosis while effector
the DNA ladder produced by cleavage of genomic DNA caspase-7 is required for apoptotic cell detachment.
between nucleosomes generates fragments with length Signalization implicates the release of a number of signal
corresponding approximately to 180 base pairs [7]. molecules, implicated in cellular damages and loss of
Phosphatidylserine (PS) exposure is another caspase- adhesion. They are also involved in the release of a number
dependent process is actively localized on the inner leaflet of of signal molecules, including receptors such as Fas/TNF,
the plasma membrane. Cytoplasmic and nuclear and glucocorticoids.
condensation, chromatin cleavage, formation of apoptotic Regulation of intracellular mediators leads to the release of
bodies, maintenance of an intact plasma membrane and effectors (proteins rich in cysteine, and nucleases leading to
exposure of surface molecules target cell corpses for survival). Molecules such as P53, Bcl2, kinases,
phagocytosis. Caspases -2, -6, -7, -8, -9 and -10 mediate the phosphatases, ceramides, proteins regulating the cell cycle
process of apoptotic cell death. and transcription factors guide to deficits in growth factors
Sequestration of cytoplasmic material within (NGF, IL-2). In summary, intracellular mediators regulate
autophagosomes leads to bulk degradation by lysosomes. debits (death, proteolysis, DNA degradation and
Autophagosomes are formed by two membranes and contain destruction). The morphological hallmarks of apoptosis
degenerating cytoplasmic organelles or cytosol allowing include DNA fragmentation and membrane blebbing
distinguishing by transmission electron microscopy between (Figures 1, 2, 5 and 6).
autophagosomes and other types of vesicles such as
endosomes, lysosomes or apoptotic blebs.

Figure 5. Immunostaining of apoptotic cells stained with anti-transglutaminase (Anti-TGase).

Odontoblasts (O) are unstained, as well as pulp cells (P)

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Figure 6. 28 days after BIO implantation, apoptotic cells are stained in the root and in the apical cell-rich zone.
Extrinsic pathway of apoptosis: Death receptors are constitute a type of cell death discriminating between
members of the tumor necrosis factor (TNF) family that programmed and fortuitous forms of necrosis.
include TNF-receptor-1 (TNFR1), CD95, death receptor 3,
Mitochondrial alterations, lysosomal changes, nuclear
TNF-related apoptosis-inducing ligand receptor-1, and
changes, lipid degradation, increase in the cytosolic
TRAIL-R2.
concentration of calcium (Ca2+) result in mitochondrial
Intrinsic pathway of apoptosis: Depends on factors overload and activation of caspase proteases. A crucial role
released from mitochondria. Caspase deficient mouse for the serine/threonine kinase RIP1 has been demonstrated.
phenotypes. Necrotic cell death is identified by the absence of apoptotic
or autophagic markers, especially when the cells undergo
Deregulation of caspases underlines human diseases
early plasma membrane permeabilization (Figures 1 and 2).
including cancer and inflammatory disorders.
Caspases are a family of genes (endoproteases) maintaining
Necrosis
homeostasis through regulating cell death and inflammation
Designate non-apoptotic accidental cell death. In the absence (6). These endoproteases hydrolyze peptide bounds in a
of phagocytosis, apoptotic bodies may lose their integrity reaction that depends on catalytic cysteine residues.
and process to secondary or apoptotic necrosis. Caspase-3, -6, -7, -8 and -9 contains a caspase-recruitment
domain (CARD), that displays a death effector domain.
This allows discriminating between programmed and
fortuitous forms of necrosis, characterized by a gain in cell Cornification
volume (oncosis), swelling of organelles, plasma membrane
Occurs in the epidermis, morphologically and biochemically
rupture and loss of intracellular contents (Figure 1). In the
distinct from apoptosis. Often referred to as “keratinization”
presence of caspase inhibitors, necroptosis or necrosis
or cornified envelope formation, it is considered as a
terminal differentiation program similar to other nucleated

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tissues. Activation of caspases cornification should be essential for cellular homeostasis. Both executioner and
regarded as a bona fide cell death program. Obtained by the initiator caspases contribute to apoptotic cell death. Necrosis
cross linking enzymes (transglutaminase types 1, 3 and 5) was once recognized as an accidental or physical cell death
acting on several substrates (loricrine, involucrine and induced by physiochemical stress [8].
SP100), the cornified envelope proteins and proteases are
OTHER TYPES OF CELL DEATHS
required for impermeability and desquamation, respectively.
A variety of cell death has been identified including:
Pyroptosis
Atypical cell death modalities, mitotic catastrophe
Proinflammatory pathway resulting from caspase-1 activity
leading to membrane breakdown is leading to Which can be accompanied by morphological alterations
proinflammatory cytokine processing. It is characterized by including micronucleation. Mitotic catastrophe can lead
pore formation in the plasma membrane, cell swelling and either to apoptotic morphology or to necrosis.
membrane disruption [7]. Pyroptosis has been defined by the
following 4 criteria: 1) Programmed by an inflammatory Anoikis
caspase activation, 2) Pore formation in the plasma This form is induced by the loss of attachment to the
membrane, 3) DNA damage with terminal deoxynucleotidyl substrate or to other cells. Excitotoxicity Wallerian
transferase dUTP nick-end labeling positivity at a lower degeneration
intensity than apoptosis, and 4) ADP-ribose polymerase
activation after the pyroptosis-mediated DNA damage. A Neurons or axon degenerates without affecting the main cell
critical role for the caspase-1-inflammasome suggests that body of the nervous system. Neurons affected by Wallerian
the inflammasome plays a crucial role in regulating degeneration remain alive.
hyperhomocysteinemia-induced endothelial dysfunction. Paraptosis, Necroptose, Entose, Oncosis, Autophagy and
Caspases 3, 6, and 7 are executioner caspases with Programmed cell death have been also identified as forms of
prodomains in the NH2-terminus that cleaves substrates cell death.

Table 1. Terms for describing dead and dying cells.

Term Characteristic(s)
Programmed cell Dependent on genetically encoded signals or activities within the dying cell; a sequence of
death potentially modifiable events leading to the death of the cell.
Mediated by a subset of caspases (Figure 1); morphology includes nuclear and cytoplasmic
Apoptosis condensation and formation of membrane-bound cellular fragments or apoptotic bodies; not
inflammatory.
Degradation of cellular components within the dying cell in autophagic vacuoles; not
Autophagy
inflammatory.
Prelethal pathway leading to cell death accompanied by cellular and organelle swelling and
Oncosis
increased membrane permeability; proinflammatory.
Necrosis Postmortem observation of dead cells that have come into equilibrium with their environment.
these are many ad-hoc variants adapted to different
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