(CANCER
RESEARCH
27 I'art 1, 1415—1421,
August 19671
Biology of the Prostate Gland: The Electron Microscopy of Cytoplasmic
Filamentous
Bodies
in Human
Benign
Prostatic
Cells Adjacent
to Can
cerous Cells1
MYRON TANNENBAUM,DAVID SPIRO,AND JOHN K. LATTIMER
Departments of Pathology and Urology, Squier Urological Clinic, College of Physicians
New York 1OOS@
SUMMARY
Multiple blocks of tissue from hyperplastic and carcinomatous
prostatic glands were studied with the electron microscope. The
surface lining cells of the hy@erplastic glands adjacent to foci of
carcinoma
frequently
contained
juxtanuclear
cytoplasmic
filamentous bodies. These filamentous bodies were composed of
filaments 350 A in diameter which were, for the most part,
regurlarly arrayed. These filamentous structures were absent in
the following cell types : (a) carcinomatous cells, (b) hyperplastic
glands of non-neoplastic
prostates, and (c) hyperplastic glands
in neoplastic prostates which were distant from the malignant
foci. The significance of these cytoplasmic filamentous structures
is at present obscure.
INTRODUCFION
In 2 recent electron microscopic studies of human prostates,
Brandes el at. (2) and Takayasu and Yamaguchi (16) reported
ultrastructural
alterations of cancer and of histologically benign
appearing glands. Some of the peculiarities that were noted in
the prostatic cancer cells are the marked pleomorphism of the
mitochondria,
the increased numbers of mitochondria,
and the
alterations in the nucleolus and nucleus. The latter observation
is commonly seen with the light microscope (19).
Two more recent ultrastructural
studies by Mao ci a!. (8) and
by Fisher and Jeffrey (4) have demonstrated
the presence of
filamentous material in the corpora amylacea as well as in the
epithelial cells of the acini containing these concretions. They
concluded, as did Moore and Hanzel (11 , 12) in a thorough
study of prostatic corpora amylacea, that they were formed from
desquamated
epithelial cells and secretions stagnated in the
acim. Fisher and Jeffrey (4) also noted that the ultrastructural
appearance of this filamentous material was indistinguishable
from that of spontaneous amyloidosis observed in man from that
experimentally induced.
In an attempt to study the mechanism of hormonal action such
as estrogens and castration on the human prostate, a biochemical
and electron microscopic survey was undertaken on tissue ob
1 This
work
has
been
supported
by
grants
from
the
John
and Surgeons, Columbia University,
New York,
tamed from fresh surgical specimens. During the fine structure
study in this investigation,
electron microscopic observations
revealed filarnentous structures to be present in the cytoplasm of
some glands and not in others. These filamentous structures are
morphologically quite different from those reported by Mao and
associates (8 , 9) and by Fisher and Jeffrey (4). The presence of
these filamentous structures as correlated with the light micro
scopic studies of these glands is the subject of this report.
MATERIALS AND METHODS
Fresh surgical material was obtained directly in the operating
room and fixed immediately upon removal from the patient. The
tissue was obtained from 20 radical prostatectomles
and 51
enucleations for benign prostatic h3pertrophy. Immediately U@Ofl
removal, the tissue was cut into smaller fragments and fixed
first in 6.25 percent glutaraldehyde
in phosphate buffer (pH
7.25—7.40) (14) for 3 to 5 hr, postfixed in 2 percent osmium
tetroxide in phosphate buffer (pH 7.3—7.4) (10), dehydrated in
ethyl alcohol, and embedded in Epon according to the method
of Luft (6). Multiple sections of Epon-embedded
material were
stained with toluidine blue for the purpose of locating both neo
plastic and adjacent normal prostatic glands. They were cut at
2 microns and examined either with the phase microscope or
with the ordinary light microscope.
The thin sections were stained first with uranyl acetate (20)
and then with lead citrate (13). They were examined with a
RCAEMIJ-3F
or a Siemens Elmiskop 1 electron microscope.
Thin sections were cut from each of 8 to 12 Epon-embedded
blocks of tissue obtained from each of 51 benign prostatic sur
gical specimens. In the case of the benign-appearing
glands, for
each 5 gm of tissue that were fixed in Bouin's fluid, one paraffin
tissue section was stained with hematoxylin and eosin for routine
light microscopy. The sections were taken from the more
posterior portions of the glands. When sampled by this method,
no foci of cancer were seen in these 51 benign surgical speci
mens.
The 20 radical prostatectomies
each provided from 20 to 45
Epon-embedded
blocks of tissue for thin sectioning and ex
hibited frank evidence of carcinoma. None of the 20 patients
received any diethylstilbesterol
before surgery.
A.
Hartford Foundation Inc. , and Irene Given and John La Porte
Given Foundation and by USPHS Grants 5-TIGM-865-04, HE
5906, and N. Y. C.-V10-75.
Received October 10, 1966; accepted April 7, 1967.
RESULTS
Foci of either well-differentiated
prostatic cancer as well as adjacent
or poorly differentiated
areas of hyperplastic pros
AUGUST 1967
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1415
Myron Tannenbaum,
David Spiro, and John K. Laltimer
tatic glands were readily identified by examination of the paraffin
sections or the thick Epon-embedded
sections (Figs. 1 , 2). The
hyperplastic glands adjacent to areas of carcinoma were similar
in appearance by light microscopy to the hyperplastic glands
present in prostates containing only benign hyperplastic glands.
By examining numerous thick Epon-embedded
sections it was
possible to further thin section for electron microscopy portions
of the prostate gland which contained either (a) foci of carcinoma,
(b) hyperplastic
@
glands adjacent to carcinoma, or (c) hyper
plastic glands at a distance from the malignant areas. When
hyperplastic
glands adjacent
to carcinomatous
areas were
examined
under the electron microscope,
intracytoplasmic
filamentous structures not previously described were observed
in the acinar surface cells. These structures measure about 1
in diameter and up to 6 @iin length and are composed for the
most part of parallel arrays of filaments (Figs. 4—6).These fila
ments are uniform in diameter, measuring 350 A, and are sepa
rated by an interval spacing of 70—80A when arrayed in
parallel (Figs. 7 , 8). The filaments often exhibit a suggestive
periodic substructure (Figs. 9 , 10). The packing of the filaments
in a plane perpendicular to their long axis has not been concisely
defined but does appear to be regular (Fig. 3). On occasion, the
filaments
comprising
these cytoplasmic
structures
are not
straight and pursue a serpiginous course (Fig. 4).
The cytoplasmic structures are found in the acinar surface
cells and not in the basal cells of the hyperplastic glands adjacent
to regions of carcinoma, and they are usually confined to the
infranuclear areas. They were observed in 16 of the 20 prostate
glands which contained carcinoma. When present, they were
found in approximately
2—8percent of the acinar surface cells
lining these hyperplastic glands. They were never observed in
the 51 specimens which contained only benign prostatic hyper
trophy but not carcinoma. They were similarly absent in nodular
hyperplastic and normal glands remote from the foci of carcinoma
in the 20 radically resected prostates. In no instance were they
observed in the cytoplasm of the carcinomatous cells.
DISCUSSION
The present ultrastructural
study of prostatic tissue revealed
intracytoplasmic
filamentous structures which were limited to
histologically benign hyperplastic acinar surface cells, adjacent
to foci of cancer. The filaments comprising these structures in
no way resembled previously described prostatic cytoplasmic
filaments
(4 , 8 , 9). The latter are considerably
smaller in
diameter. The latter also do not exhibit similar substructure and
do not exhibit parallel arrays that form discrete cytoplasmic
structures.
In order to evaluate morphologically
the filamentous struc
tures in the prostatic cells, a consideration of microtubules and
microfilaments
is warranted.
Improved
electron microscopic
technics and particularly the use of glutaraldehyde
as a fixative
have revealed minute tubelike components of the cytoplasm in
a wide variety of cells (1, 3, 5, 15, 17, 18). The characteristic
microtubule has the appearance of a straight or slightly curved
hollow structure with unvarying outer diameter, generally be
tween 150 and 250 A, and uniform inner caliber and wall thick
ness (15, 17). Further evidence of the hollow nature of micro
tubules has been recently presented in electron micrographs of
1416
tubules which have been isolated and shadowed with metal
(21). These hollow structures also extend for relatively long
distances [greater than 1.5 microns (15)] through the cytoplasm
and do not branch or interconnect with other tubules. Besides
these hollow tubular structures,
the cytoplasm of many cells
also contains microfilaments (17 , 18). These structures are gen
erally interspersed
with the straight microtubules
and, like
the latter, often extend through the cytoplasm
for several
microns without sharp bends. The microfilaments are electron
opaque, measure 80 A in diameter, and show no distinct perio
dicity along their length (17 , 18).
The filamentous structures which we have observedin certain
prostatic cells also extend, like the microtubules
and micro
filaments, through the cytoplasm (Fig. 4) for long distances.
However, morphologically,
the prostatic filamentous
bodies,
unlike the microtubules, are not hollow and have substructure
(Fi@s. 8 , 9 , 10). They are also larger than the microtubules by
50 A or more and also larger than the microfilaments by at least
270 A units. The nature and significance of the l)rostatic fila
mentous structures
demonstrated
in the present study are
obscure. Their dimensions and filamentous substructure,
how
ever, are more reminiscent of cytoplasmic inclusions (22, 23)
which have been observed in cells of frog kidney tumors. In these
tumors a virus has been incriminated as a possible etiologic agent
(7, 22, 23). Further characterization
of the nature of the pros
tatic filamentous structures will necessitate further investigation
requiring technics other than purely morphologic ones.
ACKNOWLEDGMENTS
The invaluable technical assistance of Mr. M. Rosen is grate
fully acknowledged. We are also grateful to Drs. Raffaele Lattes
and Meyer M. Melicow for their excellent help in interpreting the
histopathology
of the prostatic lesions.
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CANCER RESEARCH
VOL. 27
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Biology of the Prostate Gland
11. Moore, R. A. Morphology of Prostatic Corpora Amylacea
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AUGUST 1967
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1417
Myron
Tannenbaum,
FIG. 1. This
tive,
David Spiro, and John K. Lattimer
photomicrograph
demonstrates
benign
hyperplastic
glands
(H)
adjacent
to cancerous
glands
(Ca).
H & E, Bouin's
FIG. 2. This
is a phase
photomicrograph
of an Epon-embedded
section
cut
at 2j.s, fixed
in glutaraldehyde
and
with toluidine blue. Two to 3 layers of surface cells (5) cover a dispersed layer of basal cells (B) in a hyperplastic
focus of prostatic cancer. X 710.
1415
fixa
X 290.
CANCER
0804
, and
stained
gland adjacent
RESEARCH
to
VOL. 27
Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1967 American Association for Cancer Research.
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Biology of the Prostate Gland
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Fio. 3. This electron micrograph includes portions of the surface cells of a hyperplastic gland similar to those in Figs. 1 and 2. One
of the surface cells contains an infranuclear structure composed of filaments (F) some of which are cut in cross-section. N, nucleus.
x
12,000.
Fio. 4. A similar cell discloses a juxtanuclear
nucleus.
X 17,500.
array of filaments (F) which takes a serpiginous
course through
the cytoplasm.
AUGUST 1967
Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1967 American Association for Cancer Research.
Nuc,
1419
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5. A similar
spaced.
body
is seen
adjacent
to the
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(Nuc).
The
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(F)
are
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array
and
regularly
X 12,000.
FIG.
6.
Similar
FIG.
7.
This
1420
filamentous
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to
Fig.
demonstrates
5.
X
25,000.
another
cytoplasmic
filamentous
body
(F)
and
reveals
its
filamentous
substructure.
CANCER
X
68,000.
RESEARCH
VOL. 27
Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1967 American Association for Cancer Research.
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Fzo. 8. A filamentous
body (F) is demonstrated;
note the suggestion
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figures
are
9 and
10.
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of
the
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X
124,000 (Fig. 9) and X 270,000 (Fig. 10).
AUGUST
1967
Downloaded from cancerres.aacrjournals.org on June 16, 2017. © 1967 American Association for Cancer Research.
1421
Biology of the Prostate Gland: The Electron Microscopy of
Cytoplasmic Filamentous Bodies in Human Benign Prostatic
Cells Adjacent to Cancerous Cells
Myron Tannenbaum, David Spiro and John K. Lattimer
Cancer Res 1967;27:1415-1421.
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