[CANCER RESEARCH 49, 2525-2532, May 15, 1989]
Perspectives in Cancer Research
Cancer as a Disease of DNA Organization and Dynamic Cell Structure1
Kenneth J. Pienta, Alan W. Partin, and Donald S. Coffey
The Johns Hopkins Oncology Center and the Department of Urology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
Cancer cells develop resistance to all known natural and
synthetic drugs; normal cells do not. This resistance is a reflec
tion in part of the wide diversity of functions expressed by
cancer cells within a tumor. This variation in function (pleiotropism) is accompanied by variation in structure (pleomorphism), and together they form the basis for tumor cell heter
ogeneity. This tumor cell heterogeneity provides malignant
tumors with a tremendous biological diversity, which enables
them to succeed in a competitive environment that includes
therapeutic manipulations. The driving force in the develop
ment of tumor cell heterogeneity is thought to be genetic
instability (1-4). However, it is unknown whether cell structure
determines this instability or whether the instability of the DNA
itself produces the instability in structure (5, 6). It is well
recognized that chromatin structure can regulate DNA function
within the cell (7-11). The purpose of this "Perspectives" article
is to provide an overview of the importance of nuclear and cell
structure in DNA organization and suggest how these may be
altered in the cancer cell.
The Nuclear Matrix
How the vast array of DNA is arranged within the nucleus
in an organized fashion is difficult to comprehend. For example,
if the nucleus were magnified in size to a sphere 3 feet in
diameter, the DNA molecule would extend as a filament for
100 miles. Following replication and before mitosis, this fila
ment length would double to 200 miles. This vast amount of
DNA must be spatially organized in order to avoid any entan
glement during replication and subsequent mitosis. This could
not be accomplished by free-floating or soluble DNA but must
require a precise 3-dimensional organization and topological
considerations. The organization of interphase DNA is believed
to be accomplished by the interaction of the DNA at specific
sites to a nuclear matrix system.
The nuclear matrix is defined as the dynamic structural
subcomponent of the nucleus that directs the 3-dimensional
organization of DNA into loop domains and provides sites for
the specific control of nucleic acid intranuclear and particulate
transport (12). Conceptually, the nuclear matrix can be viewed
as the nuclear equivalent to the cytomatrix. These matrix struc
tures had also been termed "skeletal" or "scaffolding" compo
nents, until it became apparent that they exhibited dynamic
properties and were not simply rigid framework structures. The
nuclear matrix is not a single structural entity but a complex
that contains specific subcomponents such as the pore-complexlamina, residual nucleoli, and internal ribonucleoprotein parti
cles attached to a dynamic fibrous network of proteins, RNA,
and polysaccharides (13-20). The nuclear matrix structure is
essentially devoid of histones and lipids and represents less than
15% of the mass of the intact nucleus (Fig. 1).
The nuclear matrix is an important structural component in
a variety of nuclear functions reviewed in Table 1. Primarily,
Received 1/15/89; accepted 2/21/89.
1This work was supported by NIH Grants CA 15416 and AM 22000.
the nuclear matrix serves an important role in DNA organiza
tion and nuclear structure. DNA loop domains are attached at
their bases to the nuclear matrix (21, 22), and this organization
is maintained throughout both interphase and metaphase (2327). These loops are 50-150 kbp2 long and are equivalent in
size to the replicón, i.e., the amount of DNA replicated as a
unit during DNA synthesis that resides between adjacent rep
licating forks (21). There are approximately 50,000 to 100,000
of these DNA loops per nucleus. Constrained at their bases to
the matrix, the DNA loops are often supercoiled (22), which
may control in part the chromatin structure through changed
DNA topology. Topoisomerase II, one of the enzymes that
modulates DNA topology, is associated with the interphase
nuclear matrix (28, 29) and with the mitotic chromosome
scaffold (30).
The nuclear matrix also plays an important role in DNA
replication (31). The matrix contains fixed sites for DNA
synthesis (21, 32-34) located at the base of the loops. During
DNA synthesis the loop domains are reeled down through the
attached replicating complexes. When the total DNA attached
to the nuclear matrix is treated with EcoRl restriction enzyme,
a minute fraction of the DNA still remains on the matrix that
is enriched in replicative forks (35). Vogelstein et al. (22) have
been able to visualize and follow the rate of movement of labeled
DNA into these loop domains as they are replicated. Further
more, Tubo et al. (36) have reported that matrix-bound DNA
synthesis /;/ vitro continued from replication sites being used
for DNA synthesis in intact nuclei in vivo at the time of
isolation. The DNA-replicating complex located at the base of
the loop has been isolated and termed the reputase (37). The
reputase is a 24-30-nm-diameter
particle with a molecular
weight of approximately 5 million and contains at least eight
enzymes which include ribonucleoside diphosphate reductase,
thymidylate synthetase, dihydrofolate reductase, DNA methylase, topoisomerase, and DNA polymerase (37). This large
multienzyme complex appears to be under allosteric control
(38). The DNA replication fork, DNA polymerase a, and newly
replicated DNA have all been closely associated with the nuclear
matrix during DNA synthesis (32-36, 39). Earnshaw and Heck
(30) have shown that the scaffold or matrix of the metaphase
chromosome contains topoisomerase II, which is also a com
ponent of the interphase nuclear matrix during the time of
DNA synthesis (28, 29). Nelson et al. (40) have reported that
newly synthesized DNA can be covalently attached to topoisom
erase II of the interphase nucleus. With time the newly labeled
DNA moves away from the topoisomerase, which indicates that
the topoisomerase II is located at the base of the DNA loops
in close proximity to, but in the wake of, the replicating fork.
This is consistent with the observations of Noguchi et al. (41)
that topoisomerase is part of the replicase particle that forms
the replisome complex for a fixed site for DNA synthesis.
Recently, Dijkwel and Hamlin (42) have identified specific
matrix DNA attachment regions that are positioned near rep
lication initiation sites and interamplicon junctions in the am2The abbreviations used are: kbp, kilobase pairs; ECM, extracellular matrix.
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CELL DYNAMICS AND DNA ORGANIZATION
Fig. 1. Isolation of the nuclear matrix. The
nuclear matrix of a normal rat liver nuclei is
isolated by sequential extractions using nonionic detergent, brief DNase I digestion, and a
hypertonic salt buffer. These extractions re
move over 98% of the DNA, 70% of the RNA,
and 90% of the nuclear proteins resulting in a
residual structure that is essentially devoid of
histones and lipids (12-20).
EXTRACTION1.
SEQUENTIAL
DETERGENT3.
NONIONIC
HYPERtONIC SALT
NUCLEUS
PROTEIN98% PHOSPO.LIPID95%DNA70%
Table 1 Nuclear matrix is the dynamic structural subcomponent of the nucleus
that directs the functional organization of DNA into loop domains and provides
organizational sites for many of the functions involving DNA
Reported functions of the nuclear matrix
Nuclear morphology: The nuclear matrix contains structurai elements of the pore complexes, lamina, internal
network, and nucleoli which give the nucleus its overall
.^-dimensional organization and shape.
Refs.
12-20
DNA organization: DNA loop domains are attached to
nuclear matrix at their bases and this organization is
maintained during both interphase and metaphase. Nu
clear matrix shares some proteins with the chromo
some scaffold including topoisomerase II, an enzyme
which modulates DNA topology.
21-30
DNA replication: The nuclear matrix has fixed sites for
DNA replication, containing the replisome complex
for DNA replication that includes polymerase and
newly replicated DNA.
21, 22, 31-42
RNA synthesis: Actively transcribed genes are associated
with the nuclear matrix. The nuclear matrix contains
transcriptional complexes, newly synthesized hetero
geneous nuclear RNA, and small nuclear RNA. RNAprocessing intermediates are bound to the nuclear ma
trix.
47-65
Nuclear regulation: The nuclear matrix has specific sites
for steroid hormone receptor binding. DNA viruses are
synthesized in association with the matrix. The nuclear
matrix is a cellular target for transformation proteins,
some retrovirus products like the large T antigen, and
EIA protein. Many of the nuclear matrix proteins are
phosphorylated at specific times in the cell cycle.
43-46, 66-72, 74,
75, 81
NUCLEAR
MATRIX
REMOVED90%
plifÃ-eddihydrofolate domain of Chinese hamster ovary cells.
Therefore, the nuclear matrix is well positioned to play an
important structural role in the organization and biological
control of DNA eukaryotic replication. Furthermore, when
DNA viruses replicate in a mammalian cell they are also syn
thesized in association with the nuclear matrix of the host cell
(43-46). It is easy to visualize how alteration in the nuclear
matrix structures could impinge on the regulation of DNA
synthesis.
Many investigations have reported that the nuclear matrix is
associated with nuclear RNA and RNA synthesis (47-57).
Transcriptional complexes have been identified on the nuclear
matrix (47). Newly labeled heterogeneous nuclear RNA and
small nuclear RNA are enriched on the nuclear matrix (48-55).
Ciejek et al. (56) observed that 95% of unprocessed mRNA
precursor for several genes, which include ovalbumin, are as
sociated with the nuclear matrix of the chick oviduct. When the
intron portions of the primary transcript were processed out,
the mature mRNA was formed and released from the nuclear
matrix. The snRNAs are also associated with the matrix and
bind to the intron regions of the RNA transcript and may be
responsible for the attachment. Mariman and Van Venrooij
(57) reported that all RNA cleavage products and RNA proc
RNA
essing intermediates were firmly bound to nuclear matrix. Many
studies with a wide variety of genes have demonstrated that
active genes are associated with the nuclear matrix while transcriptionally inactive genes are not, providing further evidence
that the matrix may play an important organizing role in
transcriptional functions (58-65).
Several other observations indicate that the nuclear matrix is
an important modulator of nuclear regulation. The nuclear
matrix is a major site of steroid hormone receptor binding (6672). Barrack et al. (66-68) have shown that in the presence of
specific steroids, 40-60% of all nuclear steroid receptors (estro
gens or androgens) are associated with this matrix. In addition,
the nuclear acceptor for the steroid-bound receptor appears to
be part of the nuclear matrix in steroid target tissues (69, 70).
New properties of the nuclear matrix have recently been iden
tified. Fey and Penman (73) have reported that nuclear matrix
proteins, localized to the interior of the nucleus, vary in a cell
type-specific manner, suggesting that the nuclear matrix may
play an important role in development and tissue organization.
In cancer cells transformation proteins appear to be associ
ated with the nucleus, and many of these appear to be involved
with the matrix. For example, the nuclear matrix is reported to
be one of the targets for retrovirus myc oncogene protein (74,
75), adenovirus ElA-transforming protein (76), and polyoma
large T antigen (77-79). Numatrin, a nuclear matrix protein,
has been associated with the induction of mitogenesis (80).
Nuclear matrices from various cells contain binding sites for
myb proteins (81), and the nuclear matrix has been shown to
be altered during transformation (82-85). Since steroid recep
tors and transforming proteins such as the nuclear oncogene
products affect DNA function in target cells, more insight is
required into how they alter the functions of the nuclear matrix.
DNA Organization
The packaging and 3-dimensional organization of DNA
within chromatin, metaphase chromosomes, and the interphase
nucleus remain largely unsolved. Dynamic reorganization of
chromatin is visually apparent in the cell cycle from interphase
to metaphase. How the DNA loop domains are maintained
throughout these transitions of the cell cycle is still unclear.
The actual structure of these loop domains has only recently
begun to be understood. Three higher order levels of DNA
organization have been identified: nuclesomes, 30 nm chromatin fibers, and DNA loop domains. Although controversy still
exists about the exact nature of each of these structures, they
are well accepted as basic units of DNA organization. The DNA
loop domain was first proposed by Cook et al. (86) in 1976
when they suggested that loop structures are involved in the
superhelical organization of eukaryotic DNA. In 1980, Vogelstein et al. (22) reported that DNA loop domains were attached
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CELL DYNAMICS AND DNA ORGANIZATION
at their base to the nuclear matrix and that these loops were
topologically constrained by that attachment. Furthermore, the
nuclear matrix contained fixed sites for replication of DNA
loops that Pardoll et al. (21) showed were the structural equiv
alents of replicons, the basic lengths of DNA synthesized as
continuous units (87). The DNA loop domain defines a basic
unit of higher order DNA structure which is present throughout
the cell cycle in eukaryotic cells (88,89). These loops have been
estimated to be between 10 and 180 kbp pairs with an average
of 63 ±14 kbp (90). An average loop would be large enough to
contain 300 nucleosomes wound with 6 nucleosomes per sole
noid turn into a 30-nm fiber utilizing the model proposed by
Finch and Klug (91) and supported by the observations of others
(92-94). Williams et al. (95) have demonstrated that the 30nm fiber could also be constructed with crossed-linker nucleosoma! organization; however, the DNA packing ratio of both
30-nm fibers is similar. The 30-nm fiber forms the filament of
a loop, a basic structure of both interphase and metaphase
DNA. If a human diploid nucleus contains 6x10'' base pairs,
there would be approximately 100,000 of these DNA loop
domains within a single nucleus. The full organization of these
loop domains within the interphase nucleus and metaphase
chromatid has still not been elucidated. In interphase, several
lines of evidence support a model that has the DNA loops
attached to the inner portions of the nuclear matrix. Many
investigators have demonstrated that newly synthesized DNA
occurs throughout the interior of the nucleus and not just at
the periphery or lamina areas as was once believed (21, 96-99).
In metaphase, DNA loop domains are preserved in chromo
somes (100), structures devoid of nuclear envelope and lamina
proteins. The higher order structure of metaphase chromo
somes remains controversial. Several different models have
been described which include radial loop (100-102), folded
fiber (103), unit-fiber (104), spiral coil (105), and coiled-coil
(106) models. We compared the amounts of DNA in a chro
matid of the No. 4 human chromosome with the measured
chromatid dimensions at maximum condensation. DNA loops
can be packed into actual chromatid dimensions only when a
radial loop model and current concepts of higher order DNA
structure are used (See Fig. 2 and Table 2) (90). This model
features loops wrapped radically around the central axis of the
chromatid as they stack to achieve overall chromosome length.
Our analysis revealed that there would be 18 DNA loops per
radial turn of the chromatid. This 18-loop unit forms an as yet
theoretical higher order structure of DNA organization that we
have termed the "miniband" because it represents the smallest
achievable band of a chromosome (90). The miniband is equiv
alent to one full radial turn of 18 loops, each of 60 kbp, around
the central axis of the chromatid to form minibands of approx
imately 1 million base pairs of DNA (18 x 60 kbp).
The nuclear matrix provides a dynamic framework for DNA
organization. Insight into the dynamic process of the matrix
has been provided by fluorescent antibody studies of changes in
the distribution of nuclear matrix antigens at different times
during the mitotic cell cycle (107-109) (see Fig. 3). The DNA
loop domains present in the metaphase chromosome maintain
their association with the nuclear matrix in the interphase
nucleus. The telomere regions on the end of the chromosome
are attached to the peripheral lamina. As the nucleus ap
proaches metaphase, the nuclear lamina proteins are phosphorylated and have been shown to diffuse as small vesicles into
the cytoplasmic area as the nuclear envelope disintegrates (110,
111). The chromosomes, now free of the lamina, collapse into
condensed mitotic structures. At the end of telophase, the ends
The Formation
Of The Radial
Doubl« Stranded DNA
I
Loop Chromosome
XXDOOOOOOOOC
~ i,
.<=*.
Bow perlObp80
Pairs
Roho16-74068012
}2nm
.
hOnm
p1160 b
hjrntl1.20Gb
iii-.
10 nm Nucleosomes
V 6 Nucleosomes/
Turn
30 nm Solenoid
p(t*
lurnleo.ooobpl(xr
Loops
Matrix
(Topoisomerase)
loop)I.I
xio'bp<p«l«10*I.2»IO'
Miniband
n,,n,t.or.l18
so Mm•¿
loops/
MinibandPochinq
Chromosome
(Side View)
Fig. 2. Schematic of the levels of organization within a chromatid of a
chromosome. Approximately 160 base pairs (b.p.) of 2-nm DNA helix is wound
twice around the histone octamers to form the 10-nm nucleosomes. These
nucleosomes form a "beads on a string" Tiber which winds in a solenoid fashion
with 6 nucleosomes per turn to form the 30-nm chromatin filament. The 30-nm
filament forms the 60-kbp DNA loops that are attached at their bases to the
nuclear matrix structure. The loops are then wound into the 18 radial loops that
form a miniband unit. The minibands are continuously wound and stacked along
a central axis to form each chromatid. Variations in interchromosomal length are
achieved by altering the number of minibands in each chromatid. Variations in
intrachromosomal length are achieved by the winding, unwinding, and compact
ing of the minibands (90).
of the chromosome serve as an organizing center for the con
densation of lamina proteins to reform the lamina of the nucleus
(111-113). In this model some nuclear matrix structures at the
base of the DNA loops will be maintained as the core scaffolding
or matrix within the chromosome. In support of this theory,
the nuclear matrix has been shown to share many common
proteins, including topoisomerase II, with the chromosome
scaffold (23-30, 114). In the future much work will be needed
to determine how DNA loops interact with the nuclear matrix,
how the replicating complex is formed during S phase, and how
the matrix-organizing centers control and reestablish nuclear
structure. Additionally, it remains to be seen how the nuclear
matrix-DNA complex is organized and altered with the devel
opment of cancer that is associated with distorted nuclear
structures.
DNA and Chromosomal Rearrangements in Cancer
Chromosome translocations have been proposed to be a
common factor in many types of human neoplasias (115, 116).
When large translocations occur, there is also a transfer of
chromosomal banding patterns. In the model in Fig. 2, struc
tural elements of the core of the chromosome and the nuclear
matrix that anchor the DNA loops must be transferred in the
translocation process. Since sister chromatid exchanges occur
at the site of DNA replication that is on the matrix, the nuclear
matrix components may be involved in these rearrangements
(117). These types of DNA reorganization in cancer that include
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CELL DYNAMICS AND DNA ORGANIZATION
Table 2 Comparison of experimentally observed values of human chromosome 4 with those predicted from the proposed model
Human Chromosome 4 (1.15 x 10* base pairs) of DNA/chromatid
dimensionsLength
pairs/
loop
(kbp)63
±14"
dimensionsDNA
of DNA
loop
(//in)21.4*
loops/
miniband16.9±
1.9e
4
ratio12400*
Observed experimentally
Model predictionsBase
60Loop
20.4Chromosome
18DNApacking
12260Condensed
" Average value ±SEM from Refs. 163-166.
* 63,000 base pairs x 3.4 angstroms/base pair =21.4 /im.
' Average number of loops per turn on chromâtids counted from micrographs published in Refs. 100, 103, 167, and 168.
'' Length of DNA double strand divided by chromatid length (3.91 x IO4firn divided by 3.15 /im).
' Measured by Dr. G. F. Bahr, Armed Forces Institute of Pathology, Washington, DC.
INTERPHASE
(DIPLOID)
Expansion of
Chrom at in
length
(cm)3.15'
diameter
(¡um)0.85'
3.19Chromatid
0.84
CYTOMATRIX
S-PHASE
(TETRAPLOID)
PROPHASE
METAPHASE
Replication of
DNAin Loops
Slater Chromatid
Chromosome
Condensation
04-LunluLo
/MICROTUBULESA
/MICROFILAMENTSA
I INTERMEDIATE
V
FILAMENTS
/
/GLYCOCALYXX
V
INTEGRAL
PROTEINS
/
V
EXTRACELLULAR
MATRIX
COLLAGENS, LAMININS,
\
( FIBRONECTINS, PROTEOGLYCANS^
INORMALCELL!
DNA Loops'
Fig. 3. Schematic diagram of the concept of the role of the nuclear matrix in
organizing a single chromatid in the interphase nucleus. During S phase the DNA
loops replicate. During prophase the matrix separates and disengages the telomere
from the lamina. The lamins are phosphorylated and disperse into the cytoplasm
in small vesicles. The matrix attached to the DNA loops condenses during
metaphase to organize the chromosome (107-113).
gene amplification (118), sister chromatid exchange, and a
series of different subtypes of rearrangements and deletions
(119) have all been demonstrated in both animal and human
tumors. Each of these processes involves a change in the order
of arrangement of the DNA sequence and as such has been
proposed to be involved in the initiation of carcinogenesis and/
or increase progression. Any of these rearrangements which
bestows a growth advantage on the cells could cause a hyperplasia or tumor formation. If a DNA rearrangement places an
error within the genomic apparatus such that further genetic
instability ensues, a wide variety of cells would result. This type
of genetic instability may be the basis of progression and the
formation of tumor cell heterogeneity (1-5, 120). Elucidation
of the molecular events which produce this genetic instability
is critical to the understanding of cancer. It will be necessary to
define the relationship between genetic instability and the alter
ations of cell structure, which is the morphological hallmark of
cancer diagnosis, in order to further our understanding of the
cancer process. The concept of a dynamic tissue matrix system
may form a basis for this understanding.
[CANCERCELL|
Fig. 4. The tissue matrix system. The shape of the cell is dependent on
dynamic interactions of its structural components. The nuclear matrix is con
nected to the cytomatrix. The cytomatrix in turn is attached to the extracellular
matrix via the membrane matrix. Virtually every subcomponent of this matrix
system has been shown to be altered in the cancer cell (121-137).
ments form a tissue matrix that works in concert to provide a
dynamic matrix system connected throughout the tissue (121123). Fey et al. (123) proposed that the nuclear matrix is directly
linked to an intermediate filament complex that functions as a
structural unit within the cell. They provided convincing 3dimensional electron micrographs that show that the nuclear
matrix is contiguous with intermediate filaments that extended
via the desmosomes over the entire epithelial cell colony. Most
important, they have shown that the nuclear matrix-interme
diate filament system retains proteins that are specific to its
cell type and that this is unique to this substructure of the cell
system (124). Fey and Penman (125) demonstrated that tumor
promoters induce a specific morphological signature of a nu
clear matrix-intermediate filament scaffold of kidney cells. In
their studies, the nuclear matrix-intermediate filament complex
was profoundly reorganized in a specific manner after exposure
to tumor-promoting agents. These changes fit with Penman's
The Tissue Matrix System
earlier observations that modulation of cell metabolism is mod
ulated by cell shape and external surface contact (126). Many
studies have shown that much of the macromolecular metabo
lism of the cell, including DNA, RNA, and protein synthesis,
responds to changes in cell shape (127-129). Progressive loss
of shape-responsive controls may be an important factor in
tumor progression. Ben Ze'ev (130) has provided a comprehen
The nuclear matrix forms an interlocking network with the
cytomatrix that extends throughout the cell and makes external
contact with the ECM (Fig. 4). The cytomatrix is composed in
part of networks of actin microfilaments, intermediate fila
ments, and microtubules. The ECM includes the basement
membrane and ground substance of the stroma and is composed
in part of collagens, laminins, fibronectin, and proteoglycans.
Several investigators have proposed that these structural ele
sive review of the changes in the cytoskeleton associated with
cancer cells and has proposed that growth-regulated cellular
functions are regulated by signals which are transmitted
through an organized cytoskeleton that has been disrupted by
the carcinogenic process. Microfilaments, intermediate fila
ments, and microtubules have all been documented to be altered
in many transformed cells. Furthermore, it is possible that
some of the oncogene proteins are tyrosine kinases, which may
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CELL DYNAMICS AND DNA ORGANIZATION
induce some of the cytoskeleton changes that occur with trans
formation. Observations on the importance of external surface
contact and anchorage-independent growth of tumor cells have
focused much attention on the importance of the cytoskeleton
and its relationship to the ECM in tumor progression. In classic
studies, Folkman and Moscona (131) controlled the shape of
normal cells in vitro by varying the substratum adhesiveness of
the culture plates to which the cells were attached. Gospodarowicz et al. (132) observed that cell shape determines the
mitogenic response of a given cell. Additionally, the ECM has
been implicated in the control of genetic expression (133), and
Reid (134) has demonstrated that the ECM components glycosaminoglycans and proteoglycans can induce morphological
changes, induce gap junction synthesis, and regulate tissuespecific gene expression. It is well recognized that the ECM
clearly plays an important inductive function in embryonic
development and may also modulate adult cells. For example,
Cìinhaet al. (135) demonstrated that the ECM can be respon
sible for functional differentiation in development when they
showed that urogenital sinus mesenchyme induces urinary blad
der epithelial cells to form prostatic epithelial cells and acini.
Reddi and Anderson (136) observed that mature fibroblasts
underwent redifferentiation to form new chondroblasts and
chondrocytes when they were exposed to demineralized bone
collagen matrix. All of these and many other experiments
demonstrate that what a cell touches is important in determin
ing what it becomes and how it functions (121). We still know
very little about the integration and control of these cytoskele
ton and extracellular matrix processes and how the information
is transmitted to the nucleus. In summary, the cell may transmit
signals by direct mechanical linkages via the tissue matrix
system that can regulate DNA function (119-121,137). We do
know that in the cancer cell the cell matrix can be highly
dynamic. This is exemplified by cell motility.
Cancer Cell Motility
In 1940, George Gey (138) was the first to use time lapse
cinemicroscopy to study the activity of cancer cells derived from
spontaneous transformation of normal cells in tissue culture.
In 1966, Sumner Wood wondered whether cell motility was
important in the pathogenesis of cancer or simply a cell culture
artifact. He used a transparent rabbit ear chamber to study the
in vivo motility of V2 carcinoma cells (139-141). V2 carcinoma
cells migrated at velocities comparable to those of leukocytes
and 200 times faster than macrophages. Coman (142) demon
strated heterogeneity in the motility of tumor cells and sug
gested that the degree of motility correlated with histológica!
differentiation and invasive potential.
This relationship between cell motility and metastatic poten
tial is starting to be explored. Platelet-derived growth factor
(143), transforming growth factors (144), insulin (145), and
epidermal growth factor (146) have all induced motility in
normally quiescent cells. Normal cells demonstrated transient
enhanced motility after treatment with phorbol ester (147) and
ruffling, which was observed with time lapse cinematography
(148,149,150). Injection of the p21ras protein product directly
into the cell induced a transient motility (151). Recently Liotta
and Schiffman (152) have identified an autocrine motility factor
secreted by tumor cells which increases tumor cell motility.
Hosaka (153, 154) used time lapse videomicroscopy to show
differences in cell motility between various rat hepatoma cell
lines that exhibited different propensities for metastasis. Haemmerli and Strauli (155) extended this technique to the study of
human neoplastic cells and suggested that their in vitro motility
reflected their invasive behavior in vivo.
Recently, time lapse videomicroscopy and a visual grading
system have been utilized to evaluate cell membrane ruffling,
undulation, pseudopodal extension, vectorial translation, and
irregularity of the pathway of translation in normal and malig
nant cells (156, 157). The Dunning R3327 rat prostatic adenocarcinoma model provides many histologically indistinguish
able sublines of varying metastatic potential which originated
from a single animal (158). No biological, biochemical, or
morphological discriminator has previously been capable of
identifying the individual sublines or predicting their metastatic
potential (158). Mohler et al. (156, 157) demonstrated that five
sublines and normal rat prostate cells could be identified by a
visual grading system of cell motility. More recently, Partin et
al. (159) have developed a new system for quantitating all
aspects of cell motility. This new quantitative method was able
to correlate cell motility changes with an increase in metastatic
ability in the Dunning tumors (159). Furthermore, Partin et al.
(160) demonstrated that motility and metastatic potential can
be induced in the Dunning tumors by transfection and expres
sion of the V-Harvey-ros oncogene. Thus motility of individual
cancer cells in vitro has been shown to be sufficiently character
istic to allow accurate assessment of their metastatic potential
in vivo. Current pathological grading systems depend upon the
appreciation of cytological and architectural features of dead,
fixed histological sections of malignant tissues. A grading sys
tem of motility of live cancer cells may better predict the
behavior of a live tumor system.
Tensegrity
The types of mechanical systems that can transfer informa
tion within a cell are just beginning to be resolved. Ingber and
Jameson (161) have suggested a tensegrity model to explain
how cells composed of structural elements may be capable of
such information transfer. Tensegrity was defined by Buckminster Fuller in 1948 as a structural system composed of
discontinuous compression elements connected by continuous
tension cables, which continually interacted in a dynamic fash
ion. This structure allows great motility as each part is in
coupled equilibrium so that mechanical forces can be trans
ferred throughout the entire system. Recently, Dennerll et al.
(162) have observed and measured a tension and compression
system in neuntes. They suggest a complimentary force inter
action between an actin network under tension and the microtubule network under compression. A tension-derived tenseg
rity structure may be a more appropriate way to view cell
structure than a rigid scaffold framework system. These changes
progress to a cell with increased motility and an ability to
metastasize.
Conclusion
The pioneering biophysicist Aaron Kachalsky stated in 1962
that "life may be defined as a chemomechanical engine." The
highly motile yet structured cancer cell may also be viewed as
a chemomechanical engine in which structure and function are
intimately interrelated. Disruptions or changes in the matrix
system may help explain genetic instability and tumor cell
heterogeneity. Our present view of this process is depicted in
Fig. 5. It is now obvious that cell transformation is a multistep
process involving genomic changes that can involve oncogenes
acting at different sites within the cell. These changes progress
2529
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CELL DYNAMICS AND DNA ORGANIZATION
NORMAL
TRANSFORMATION
-
PROGRESSION
21.
22.
CELL SIGNALING
ONCOGKNBS
23.
ACTING AT
CYTOSDLITON.
PLASMA MEMBRANE
(e.g. r»s)
STRUCTURAL INSTABILITY
DNA INSTABILITY
TUMOR CELL HETEROGENEITY
IMMORTALIZATION
24.
25.
/ /
j /
MOTILITY
(„eta.taaes)
26.
27.
Fig. 5. The transformation process. Transformation from a normal cell to a
malignant one appears to involve multiple steps. These steps are usually consid
ered in terms of "initiation" and "progression." This process may be viewed at
the mechanism/site of action of the different oncogenes involved in tumor
progression. One class of the oncogenes, those acting on the nucleus, e.g., myc,
alter the structural stability of cells. This leads to immortalization and concomi
tant DNA and structural instability. A second class of oncogenes, those acting on
the periphery of the cell, e.g., ras, induce motility into the cell and may impart
the ability to metastasize.
28.
29.
30.
to a cell with increased motility and an ability to metastasize.
Understanding this carcinogenic process will require a more
complete knowledge of the interlocking matrix systems that
extend from the extracellular matrix to the DNA and that
govern cell shape and function.
31.
32.
33.
34.
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Cancer as a Disease of DNA Organization and Dynamic Cell
Structure
Kenneth J. Pienta, Alan W. Partin and Donald S. Coffey
Cancer Res 1989;49:2525-2532.
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