[CANCER RESEARCH 30, 2791-2795,
November 1970]
The Ultrastructure of Bowen's Disease: Nuclear and Nucleolar Lesions
J. Kara'sek,1 K. Smetana, W. Oehlert, and B. Konra'd
Departments of Pathology and Dermatology, J. E. Purkyne University, Brno, Czechoslovakia [J. K., B. K.]; Department of Pathology,
University of Freiburg, Freiburg, Germany [W. E.]; and Laboratory for Ultrastructure Research of the Czechoslovak Academy of Sciences,
Prague, Czechoslovakia [K. S.J
SUMMARY
Histologically typical skin lesions of 10 patients with
Bowen's disease were examined by means of the usual
electron microscopic procedures to provide more information
on the fine structure of nuclei and nucleoli in giant cells.
Nuclei of giant cells were characterized by the presence of
an increased number of interchromatinic and perichromatinic
granules. In addition, dense globular structures about 500 to
1500 A in diameter were noted in association with chromatin. The spherical clusters observed in interchromatinic
areas at the condensed chromatin were composed of particles
about 300 A in diameter.
Most nucleoli in giant cells were either small and compact
or with nucleolonemas. Some small nucleoli of spherical
shape were characterized by segregation of fibrillar and
granular ribonucleoprotein components. Ring-shaped nucleoli
as well as nucleoli with unusual organization of nucleoprotein structures were found only in small numbers.
The ultrastructural appearance of nucleoli in some giant
cells suggested a low rate or inhibition of the synthesis of
nucleolar RNA in these cells.
INTRODUCTION
The presence of giant cells in the altered epidermis is one
of the main histopathological
and diagnostic features of
Bowen's disease. The nuclei of these cells are usually very
large, irregularly shaped and hyperchromatic (1). Their fine
structure, however, is less known (7, 11, 14).
For more information on the fine structure of nuclei
including nucleoli in giant cells of altered epidermis of
Bowen's disease, these structures were studied electron
microscopically. The results have shown that nuclei of giant
cells contained numerous interchromatinic
and perichro
matinic granules as well as atypical spherical and globular
structures. In addition, only a relatively low number of
nucleoli in giant cells seemed to be of usual ultrastructural
appearance compared with the cells of normal epidermis.
MATERIALS AND METHODS
Skin biopsy specimens from 10 untreated patients with
clinical and histological lesions typical of Bowen's disease
1Fellow of the Alexander von Humboldt Stiftung.
Received April 7, 1970; accepted July 8, 1970.
were used for electron microscopic studies. The lesions were
excised under minimal local procaine anesthesia. Small
blocks cut from each sample were fixed immediately in 2.5%
glutaraldehyde at 4°for 20 hr. Then the blocks were washed
twice in 0.15 M phosphate buffer at room temperature for 1
to 2 hr. After embedding in Epon, areas for ultrathin
sectioning were chosen after previous examination of "thick"
sections stained with toluidine blue and Azur II. Ultrathin
sections stained with uranyl acetate followed by lead citrate
were observed with a Siemens-Elmiskop IA electron microscope.
RESULTS
The cytoplasmic structural appearance of giant cells of
Bowen's disease did not differ in general from that described
in previous studies (11, 14).
The ultrastructural morphology of nuclei and nucleoli was
very similar in both mononucleated and multinucleated giant
cells.
The Nuclear Ultrastructure of Giant Cells. The nuclear size
in ultrathin sections was variable. The shape of the nuclei
appeared
irregular,
and numerous
deep cytoplasmic
invaginations into nuclei were observed (Fig. 1). As in
normal epidermal cells (10), the nuclear chromatin structure
in giant cells was composed of fibrils which were present in
large amounts ("condensed chromatin") along the nuclear
membrane, around nucleoli, and in chromocenters (Fig. 2).
At the margin of chromatin structures including nucleolusassociated chromatin, perichromatinic granules were more
frequent in the nuclei of giant cells (Figs. 2 and 3), in
comparison with normal epidermal cells (10). The size of
perichromatinic granules varied considerably between 300
and 600 A. Interchromatin
areas contained numerous
"dense" interchromatinic granules which occasionally formed
clusters (Fig. 4). Nuclear bodies were frequent; their ultrastructure in giant cells of epidermal lesions of Bowen's
disease has been described in the previous study (7). In
addition
to the structures mentioned
above, unusual
spherical and globular structures were found in the nuclei of
a few giant cells. Spherical structures were represented by
clusters of particles, about 300 A in diameter, which were
usually observed at the margin of condensed chromatin (Fig.
2). Globular structures (formations) 500 to 1500 A in
diameter were located in clumps of condensed chromatin
(Fig. 5).
The Nucleolar Ultrastructure in Giant Cells. In ultrathin
sections, the number of nucleoli ranged between 1 and 5
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2791
/. Kardsek, K. Smetana, W. Oehlert, and B. Konrad
(average 2 to 3) per nucleus. The ultrastructural morphology
of nucleoli varied not only in various nuclei but also in the
same nucleus (Fig. 1).
Most nucleoli in giant cells were either small and compact
[about 30% of nucleoli (Figs. 2 and 6)] or they were large
and composed of nucleolonemas (about 40% of nucleoli).
Some small and compact nucleoli of spherical shape were
characterized by a distinct segregation of nucleolar compo
nents (Fig. 6). Some nucleoli with nucleolonemas contained
large filamentous areas (Fig. 7). The density of these struc
tures was lower than in nucleolar fibrillar ribonucleoprotein
components.
Ring-shaped nucleoli (16) were present only in limited
numbers of nuclei. They were characterized by the presence
of ribonucleoprotein
structures in the nucleolar peripheral
part which formed a shell surrounding a central light core. In
a few nuclei, nucleoli with unusual organization of ribo
nucleoprotein structures were observed (Fig. 8). Such nu
cleoli were composed of a central mass containing mainly
fibrillar components, which was surrounded by a light circu
lar zone and peripheral ribonucleoprotein shell. It is evident
that the section across the top of such a nucleolus would
present an appearance similar to that of the ring-shaped one.
Lacking serial sections, we cannot ascertain whether each of
both last-mentioned nucleoli represents a different type of
nucleolus.
DISCUSSION
This study provides additional information on the ultrastructural morphology of giant cells in the epidermal lesions
of patients with Bowen's disease. However, in contrast to the
previous studies (7, 11, 14), the present observations deal
with nuclear and nucleolar ultrastructural morphology.
The results of the present study have shown that the
ultrastructural morphology of many nucleoli in giant cells of
Bowen's disease is greatly similar to the nucleolar changes
produced by the inhibition of the RNA synthesis. The
segregation of ribonucleoprotein
components observed in
small and compact nucleoli of giant cells resembled the
nucleolar segregation induced by inhibition by some antimetabolites of DNA-directed RNA synthesis (3, 5, 15, 17).
In addition, ring-shaped nucleoli found in some giant cells
were similar to those observed in cells with a very low rate
of RNA synthesis, which was produced either by the cell
maturation or by some inhibitors of RNA synthesis (12, 13,
16). Thus, the ultrastructural
morphology of nucleoli in
numerous giant cells of Bowen's disease seems to indicate an
inhibition or a low rate of nucleolar RNA synthesis in these
cells. On the other hand, the synthesis of extranucleolar
RNA in such cells does not appear to be altered since the
number of perichromatinic
granules was very high and
increased, in comparison with normal epidermal cells. A
similar high number of perichromatinic
granules was
observed in various cells after the treatment with actinomycin D (16), aflatoxin, and lasiocarpine (9) at doses which
presumably block nucleolar RNA synthesis but do not
inhibit the synthesis of other species of RNA (8,9).
2792
Other nuclear as well as nucleolar characteristics and
abnormalities cannot be explained, although it cannot be
excluded that they belong to the category of nonspecific
changes observed in tumor cells (3, 4, 6). However, giant
cells of Bowen's disease are also considered as degenerative
cell forms (2). The nucleolar ultrastructure in numerous
giant cells indicating a low rate or inhibition of RNA
synthesis in nucleoli, as mentioned above, does not exclude
such a possibility and may indicate the beginning of regres
sive changes connected with cell degeneration.
ACKNOWLEDGMENTS
We gratefully acknowledge the helpful criticism of Dr. R. Simard
during the preparation of this paper. We also thank Miss. J. Mazalova,
Mr. J. Blumajer, Mr. O. Hornák,and Mrs. A. Sazma for their technical
assistance.
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infrastructure ofBowen 'sDisease
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Fig. 1. Skin lesion of Bowen's disease with giant cells. N, nuclei; arrows, nucleoli. X 5,600.
Fig. 2. Nucleus of a giant cell with a large number of perichromatinic granules (arruws) and a spherical structure (pointer) composed of small
particles. C, chromocenter;^, nucleolus. X 30,000.
Fig. 3. Perichromatinic granules (arrows) in the nucleolus-associated chromatin (pointer). N, nucleolus. X 33,000.
Fig. 4. Nuclear interchromatinic granules (pointer). N, nucleolus. X 30,000.
Fig. 5. Nuclear globular formations (pointer). N, nucleolus. X 50,000.
Fig. 6. Small compact nucleolus with segregation of fibrillar (/•')
and granular (G) components. C, cytoplasm. X 40,000.
Fig. 7. A large and round nucleolus with nucleolonemas and large filamentous areas (/). X 23,000.
Fig. 8. Nucleolus with zonal distribution of nucleolar ribonucleoprotein components, A, central area with ribonucleoprotein components; C,
cytoplasmic invagination into the nucleus. X 40,000.
NOVEMBER
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2795
The Ultrastructure of Bowen's Disease: Nuclear and Nucleolar
Lesions
J. Karásek, K. Smetana, W. Oehlert, et al.
Cancer Res 1970;30:2791-2795.
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