Invasion of Skeletal and Smooth Muscle by L1210 Leukemia1
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[CANCER RESEARCH 27 Part 1, 2159-2178, November 1967]
Invasion of Skeletal and Smooth Muscle
by L1210 Leukemia1
DAVID BRANDES,2 ELSA ANTON, AND BRIAN SCHOFIELD
Departments of Pathology, The Johns Hopkins University School of Medicine, and Baltimore City Hospitals, Baltimore, Maryland
SUMMARY hensive study on the LI 210 leukemia in our laboratory, were
undertaken to determine whether electron microscopic and histo-
Invasion of skeletal and smooth muscle fibers by L1210 leu-
kemie cells was studied by electron microscopy and histochemis- chemical examination of invaded muscle would provide further
information on the events at the host-tumor interzone that would
try. In the host-tumor interzone, degenerative changes were first
shed some light on specific factors related to invasiveness.
detected in the tumor cells, and their cytoplasmic components,
especially ribosomes, appeared free in the interstitial spaces.
Many of the images strongly suggested that such tumor cell MATERIALS AND METHODS
components may have been taken up by the muscle cells, es The solid and ascitcs forms of the LI 210 leukemia were carried
pecially in areas of fusion between normal and malignant ele in DHA/2 mice weighing approximately 20 gm. In the case of the
ments. In some areas, the presence of tumor cell material seemed solid form, 0.1 ml of a saline suspension (1:10) of the spleen from
to exert a stimulatory effect on the normal structures, reflected a leukemic donor was injected intramuscularly. For the ascites
by hyperplasia of cell organelles rather than cellular multiplica form, a 1:10 saline suspension of ascites fluid from a leukemic
tion. Degenerative changes and lysis of muscle fibers occurred donor was prepared, and 0.1 ml was injected intraperitoneally.
predominantly in areas of tumor cell degradation, rather than in Twenty-four animals were injected in each group and were sacri
the presence of healthy tumor cells, indicating that products ficed on the fifth and sixth day after inoculation.
derived from altered tumor cells may play an important role in The local tumor developed in the skeletal muscle at the site of
neoplastic invasiveness. Our results also indicate that in the case inoculation of the solid form, and areas of infiltration of the
of this particular tumor, lysosomes probably do not play a role intestinal wall and the diaphragm in the mice injected with the
in the destruction of normal structures. ascites cells were rapidly removed. All tissues were fixed in 3%
glutaraldehyde in cacodylate buffer (24) for 2 hours at approxi
INTRODUCTION mately 4°Cand washed and stored overnight in the same buffer.
The actual mechanisms of invasion and replacement of normal For electron microscopy, the tissues were dehydrated in alcohols,
structures by malignant cells has given rise to much speculation embedded in Epon 812, and ultrathin sections were cut with the
in the past, based greatly on inferences derived from histologie LKB Ultrotome or with the Porter-Blum microtome. The pro
observations (6). The invasive capacity of neoplastic cells has cedures for the histochemical preparations, both at the light and
been attributed to their power of progressive and rapid multipli electron microscopy levels, have been described in detail in a
cation, to their motility and phagocytic capacity, to the elabora previous paper (9). The grids were counterstained with uranyl
tion of lytic or toxic products, or to the loss of growth restraint acetate and were observed and photographed with the RCA
normally exercised by cells on each other. These classic ideas have EMU-3F electron microscope.
been reviewed by Willis (33) and later by Berenblum (6).
More recently, the mechanisms of tumor invasion have been RESULTS
reexamined in animal experimental studies and with the aid of
elaborate biologic models. Many of the results have pointed to The Fine Structure and Histochemislry of Leukemic
the importance of proteolytic enzymes and toxic substances Cells
from tumor cells as factors which may facilitate the destruction
of normal elements and the progression of the neoplasia [see The L1210 leukemic cells were characterized by the presence of
Vasiliev (31, 32) and Sylvan (27, 29) for reviews]. large nuclei occupying a great portion of the cell (Fig. 1). Abun
The investigations reported here, which form part of a compre- dant free ribosomes were found in the cytoplasm, but very few
cisternae of the rough endoplasmic reticulum could be seen in
these cells. Prominent Golgi complexes and virus i¡articleswere
1This investigation was supported by (jrants CA 08518 from seen in most cells. The ]¡articleswere located exclusively in the
the National Cancer Institute, and HD 00042 from the National cytoplasm, either in the matrix or within cisternae of the endo
Institute of Child Health and Human Development, NIH, USPIIS.
2Recipient of a Career Development Award, K3-CA-21,756-04. plasmic reticulum. On the basis of their morphologic appearance
National Cancer Institute, USPHS. and outer diameters, the viruses have been designated as A
Received April 11, 1967; accepted July 11, 1U67. particles in accordance with the proposed classification of Dalton
NOVEMBER 1967 2159
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David Brandes, Elsa Anton, and Brian Schofield
(12).3 A more detailed description of the L1210 leukemie cells of the invaded muscle. In some instances they contained myelin
and of the virus particles characteristic of this neoplasia are to be bodies which seemed to be derived from degenerating cell or-
found in two recent publications (7, 8). ganelles such as mitochondria and ribosomes (Figs. 10, 11). The
In light microscope preparations stained by the acid phos- hyperplastic areas frequently appeared as protrusions which
phatase technic for the demonstration of lysosomes (Fig. 2), the contained predominantly abundant ribosomes (Fig. 11) or hyper-
leukemie cells appeared predominantly negative, but a strong trophic mitochondria and hyperplastic Golgi elements (Figs. 12,
positive reaction was observed in macrophages. The electron 13). Degenerative changes were detected in many of the marginal
histochemical preparations (Pig. 3) confirmed the paucity of hypertrophie mitochondria (Fig. 13). The membranes of the
lysosomes in leukemie cells, which usually contained but a few cristae lost their sharpness, and in many instances the mito
of those particles. In the preparations thus far examined, neither chondria were occupied by interlacing bands of amorphous ma
lysosomes nor acid phosphatase activity could be seen in the terial. Structures which seemed to represent residues of de
host-tumor interzone. generating mitochondria also appeared in these areas.
Progressive alterations in skeletal muscle during leukemie cell
The Host-Tu mor Interzone invasion, which appeared directly related to their breakage and
replacement of muscle fibers by tumor cells, were observed.
Light Microscopy. The degree of infiltration and destruction The sarcolemma disappeared, and the muscle showed a frayed
of skeletal muscle and smooth muscle on the fifth day after trans irregular surface (Fig. 14). In more advanced stages the surface
plantation of the leukemia are illustrated in light micrographs showed a frank moth-eaten appearance and the myofibrils had
(Figs. 4, 5). In both cases, the neoplastic cells have infiltrated begun to disintegrate (Fig. 15). The loss of muscle substance gave
and widened the spaces between the muscle fibers, many of which rise to the formation of lacunar spaces, and as this process ad
appear as isolated fragments. vanced deeper into the muscle, the fibers became thinner and
Electron Microscopy. Skektal Muscle Invasion. Before the irregular (Fig. 16) and showed areas of constriction that appeared
onset of recognizable changes, the plasma membrane of the to lead to actual fragmentation and disintegration (Figs. 17, 18).
leukemie cells, as well as the sarcolemma of the muscle fibers, Intestinal Smooth Muscle Invasion. As in the case of skeletal
appeared as continuous dense structures (Fig. 6). The sarcolemma muscle invasion, some of the smooth muscle fibers showed signs
was externally coated by a basement membrane, and a moderate of stimulation reflected in enhanced pinocytosis, mitochondrial
amount of connective tissue fibers were seen between the leukemie aggregates, and hyperplastic Golgi complexes (Fig. 19). Abundant
cells and the muscle. Aggregates of ribosomes, apparently de clusters of free ribosomes in the interstitial spaces were observed
rived from leukemie cells, were frequently detected in the inter in the host-tumor interzone, and very frequently the hyperplasia
stitial spaces t>etween leukemie cells and muscle, before any of cell organelles occurred in the vicinity of the areas with extra
morphologic alterations occurred in either cell type (Fig. 7). cellular ribosomes (Fig. 20).
Progressive alterations in leukemie cells appeared to precede Degenerative changes in the leukemie cells were also seen to
the changes in muscle fibers. Partial or almost complete disap precede alteration in the smooth muscle fibers. In the neoplastic
pearance of the plasma membrane occurred in many of the leu cells (Fig. 21), the perinuclear cisternae appeared very dilated,
kemie cells (Fig. 8), and their cytoplasmic content, especially the mitochondria were swollen, and most of the cristae had dis
ribosomes, appeared free in the interstitial spaces in close ap appeared. The plasma membrane was no longer visible, and the
position to the surface of the muscle fibers. Several degenerative various cytoplasmic components of the leukemie cells, especially
changes including pycnosis and frank necrobiosis (Fig. 9), were the ribosomes appeared free in the interstitial spaces. In some
observed in leukemie cells in the vicinity of morphologically in areas, the plasma membrane of the smooth muscle cells appeared
tact muscle fibers, but virus particles could still be detected interrupted, and the fusion of host-tumor cell cytoplasm became
among the debris of the neoplastic cells (Fig. 9, inset). established (Fig. 21).
Several changes were observed at the surface of the muscle The areas of tumor cell-muscle fusion seemed to constitute a
fibers in areas where the plasma membrane of the leukemie cells "no man's land" with cell organelles "intruding" into both sides
became discontinuous or disappeared, or in the presence of debris of the line, for mitochondria and myofibrils were seen to lodge
from necrobiotic leukemie cells. Increased micropinocytotic simultaneously in muscle and leukemie cells (Fig. 22). Pinocytotic
activity and the development of areas of hyperplastic marginal vacuoles, externally coated or containing ribosomes apparently
sarcoplasm were among the earlier detectable changes in muscle derived from those present in the interstitial spaces, were ob
fibers which otherwise appeared normal. served in muscle cells, and in areas of fusion, tumor cell ribo
The micropinocytotic vesicles appeared arranged in rows along somes appeared to gain access into muscle fibers (Fig. 23). Initial
the muscle surface and contained structureless material of low- lytic changes were observed in such areas (Fig. 23), and patterns
electron density (Figs. 8, 10, 11). The contents of the areas of of disintegration of the normal structures in contact with altered
hyperplastic marginal sarcoplasm varied in the different portions leukemie cells were similar to those described in relation to the
invasion and replacement of the skeletal muscle by malignant
3 It has been proposed, more recently, that the particles in the cells.
cytoplasmic matrix be designated as ¡ntracytoplasmic A particles,
The apparent stimulative effect of tumor cell material on nor
ami those in the eiuloplasmic reticulum as intracisternal A parti mal structures appeared to be exerted also on small blood vessels
cles. Suggestions for the Classification of Oncogenic RNA Viruses. present in the areas of tumor cell invasion. In areas containing
Informal Meeting, New York, February 9, 19fi(i (J. Nati. Cancer tumor cell debris, the cytoplasm of vascular endothelial cells
Inst,, 37: 395-397, 1966). appeared broadened and contained abundant cell organelles,
2160 CANCER RESEARCH VOL. 27
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Invasion of Muscle by LI210 Leukemia
particularly ribosomes (Fig. 24). The luminal surface was thrown At the level of resolution of the electron microscope, however,
into complex projections and folding's, indicative of intense the gap between tumor cells and skeletal muscle was considerably
pinocytosis, and hyperplasia of the Golgi apparatus and of the larger, and the alterations of the normal structures appeared to
rough endoplasmic reticulum was also prevalent (Fig. 25). occur in areas where tumor cells were breaking down, rather
than in relation with vigorously multiplying leukemic cells.
DISCUSSION Noi (21) failed to detect morphologic or histochemical changes
in kidney cells situated in front of or even in direct contact with
The present results have indicated that during invasion of growing tumor cells, and he questioned the idea that extracellular
skeletal and smooth muscle, alteration of the leukemic cells, proteolysis or other destructive processes in normal tissues might
including necrobiosis, occurs before the onset of degenerative precede tumor invasion.
changes in the normal tissues. Those studies, however, were based on light microscopy ob
Partial or complete disappearance of the plasma membranes servations, and it is possible that the author failed to detect the
of the neoplastic cells exposed the surface of the muscle fibers presence of tumor cell debris in areas of invasion and the initial
directly to the cytoplasmic components of the leukemic element subcellular changes described in this study.
and particularly to the ribosome aggregates. Alteration and lysis of skeletal and smooth muscle fibers were
In such areas, increased pinocytotic activity was detected at also detected in areas not immediately in contact with growing
the surface of the muscle fibers, which may indicate the uptake tumor cells, suggesting that substances extruded from altered
of material derived from the altered leukemic cells. More direct neoplastic cells may have acted via the interstitial fluid. Lytic
evidence pointing to interchange of material between normal and changes have been observed at distances greater than 1(X)ß
neoplastia elements was derived from the presence of numerous from tumor cells, and on the basis of histochemical and biochem
areas of fusion between both cell types, in which cell organelles ical evidence, this process has been referred to as "extracellular
appeared to protrude from one cell type into the other. proteolysis" (25, 26), indicative of local abnormal characteristics
The presence, in the interstitial spaces, of material derived of the interstitial milieu prevailing in and around tumors (27).
from altered leukemic cells appeared at first to exert a stimu Increased proteina.se activity at the host-tumor interzone, and
lative effect on the muscle fibers and also on the small blood possible activation of these enzymes through imbalance of regu
vessels present in the areas of tumor cell invasion. In the skeletal latory mechanisms, have been related to the destructive capacity
muscle fibers, the apparent stimulative effect was reflected in the of neoplastic cells (27, 29). The possibility that cathepsin and
development of areas of marginal sarcoplasmic hyperplasia, other lysosomal enzymes may be involved in tissue breakdown
increase in the number and size of the mitochondria, and hyper in areas of tumor invasion has been discussed by Pearse and
trophy of the Golgi elements. Aggregates of mitochondria and Hess (22), but these authors failed to detect high acid phospha-
hypertrophy of the Golgi components were also observed in the tase activity in tumor cells, except in stromal macrophages.
smooth muscle fibers, and in some instances, both types of hyper- Similar findings are reported here, where our preparations at the
plastic organelles appeared in close association. The stimulative light and electron microscopic levels showed very few lysosomes
effect on blood vessels occurred in areas where the interstitial in the leukemic cells and no acid phosphatase activity in areas
spaces were occupied by cytoplasmic products derived from of muscle tissue breakdown.
neoplastic cells. The cytoplasm of the capillary endothelial cells Structural defects in the organization of tumor-cell plasma
appeared broadened and studded with ribosomes, the endoplas- membrane and increased permeability of this structure (1-3, 5,
mic reticulum and Golgi appeared unusually prominent, and the 27, 28), have been held res]x>nsible for the release of cytoplasmic
luminal surface was thrown into elaborate infoldings and finger- material including enzymes, but these studies tend to indicate a
like projections which seemed to be actively engaged in the more severe process, that is, the actual disappearance of the cell
formation of pinocytotic vacuoles. membrane in the more peripheral tumor cells (type A cells of
A stimulatory effect of tumor cells or their products on normal Sylvénand Malmgren) (29).
tissues, including blood vessels and stroma, has been reported in It is possible that some of the cytoplasmic constituents of the
numerous instances, both in vivo (4, 14, 17, 23, 30, 32), and in leukemic cells, penetrating the normal structures in areas of
vitro (17-19, 23). In such instances, however, tumor cells or fusion or by pinocytosis may have been responsible for the en
their extracts stimulated the proliferation of normal cells, but in suing degenerative changes and final lysis of the muscle fibers.
our case only hyperplasia of cell organelles was encountered, The marginal sarcoplasm of striated fibers as well as in some of
which may have been due to the nondividing nature of the the smooth muscle fibers contained more ribosomes than are
tissues examined in this study. usually found in normal conditions. Many of our electron micro
What the biologic changes are that occur at the host-tumor graphs seemed to indicate that some of the cell ribosomes were
interzone that lead from a condition of stimulation to one of present in the interstitial spaces, and that incorporation into the
degradation and lysis of normal structures are not known with muscle fibers may have occurred by pinocytosis or at the areas
certainty, but have attracted much interest and have been the of fusion of malignant and normal cells.
subject of extensive experimental studies (see Ref. 11 for reviews). The material extruded from tumor cells which had lost their
In our light microscopic preparations, the areas of muscle cell plasma membrane may have served other purposes than to act
invasion were packed with tumor cells, and it appeared that as a toxic agent involved in the lysis of normal elements in the
compression alone, as suggested in classic textbooks of pa course of invasion. As suggested by other authors (16, 31), the
thology, may have been a cause in the atrophy and breaking down material released from the tumor cells may have contained a
of the normal structures. conditioning or growth-stimulating factor required for the suc-
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David Brandes, Elsa Anton, and Brian Schofield
cessful establishment of the isologous tumor graft, or may have 8. Brandes, D., Schofield, B. H., Slusser, R., and Anton, E. Stud
served as a source of nutritional material for other tumor cells. ies of L 1210Leukemia. I. Ultrastrueture of Solid and Ascites
Cells. J. Nati. Cancer Inst., 37: 467-485, 1966.
It may therefore be postulated that cell products, including en
zymes and cytoplasmic organelles, liberated by the tumor cells 9. Brandes, D., Sloan, K. W., Anton, E., and Bloedorn, F. Effect
of X-Irrudiation on the Lysosomes of Mice Mammary Gland
may have fulfilled several functions such as stimulation of normal Carcinoma. Cancer Res., 27: 731-746, 1967.
elements, conditioning the establishment of the tumor graft and 10. Braun, W. Effects of Cell Components on Cell Multiplication
served as a source of nutrition for other neoplastic cells. Such a and Changes in Cell Populations. In: M. J. Brennan and
feedback-like mechanism as stimulation of one Å“il type by W. L. Simpson (eds.), Biological Interactions in Normal and
products derived from the breaking up of other cell types has Neoplastic Growth; Henry Ford Hospital International Sym
been considered as an extremely economic intercellular control posium, pp. 187-197. Boston: Little Brown and Co., 19(i2.
mechanism (10). A lytic action on the invaded structures by 11. Brennan, M. J., and Simpson, W. L. (eds.). Biological Interac
activated enzymes or through the toxic effect of tumor cell tions in Normal and Neoplastic Growth; Henry Ford Hospital
components would then constitute one facet of the many prop International Symposium. Boston: Little Brown, and Co.,
erties attributed to the breakdown products derived from tumor 19IÌ2.
12. Dalton, A. J. Micromorphology of Murine Tumor Viruses and
cells. of Affected Cells. Federation Proc., HI: 936-941, 1962.
Some of the alterations observed in skeletal and smooth muscle 13. Engel, W. K. Mitochondrial Aggregates in Muscle Disease. J.
fibers in the areas of tumor cell infiltration greatly resembled Histochem. Cytochem., 12: 40-48, 1904.
changes described in various muscle diseases. 14. Foulds, L. The Histológica! Analysis of Tumors: A Critical
Mitochondrial aggregates, such as those seen in both types of Review. Am. J. Cancer, 39: 1-24, 1940.
fibers, have been described in patients with myotonia congenita, 15. Gustafsson, R., Tata, J. R., Lindberg, O., and Ernster, L.
paramyotonia congenita, hypokalemic periodic paralysis, and The Relationship Between the Structure and Activity of Rat
adynamia episodica hereditaria (13). Hypertrophy and increase Skeletal Muscle Mitochondria After Thyroidectomy and
Thyroid Hormone Treatment. J. Cell Biol., 26: 555-578, 1905.
in mitochondrial population in perinuclear and subsareolemmic
regions such as those seen in this study have also been observed 10. Klein, G., and Klein, E. The Evolution of Independence From
Specific Growth Stimulation and Inhibition in Mammalian
in rat skeletal muscle after thyroidectomy and thyroid hormone Tumor Cell Populations. Symp. Soc. Exptl. Biol., //: 305-328,
stimulation (15). Areas of marginal sarcoplasmic hyperplasia 1957.
containing abundant ribosomes such as seen in this study have 17. Levi-Montalcini, R., and Angeletti. P. U. Considerations on a
also been described in striated muscle in the case of trichinosis Nerve Growth Promoting Activity of some Tumors. In: M. J.
(20). Brennan and W. L. Simpson (eds.), Biological Interactions in
In our studies, these changes appeared to be localized ex Normal and Neoplastic Growth; Henry Ford Hospital Inter
clusively in the areas of tumor cell infiltration, thus indicating a national Symposium, pp. 225-237. Boston: Little Brown and
local reaction to the invading neoplastie celLs rather than a Co., 1902.
systemic disease as in the case of most of the above conditions. 18. Ludford, R. J., and Barlow, H. The Influence of Malignant
Cells Upon the Growth of Fibroblasts in vitro. Cancer Res.,
4: 094-703, 1944.
ACKN OWLEDGMKNTS
19. Ludford, R. J., and Barlow, H. Sarcomatous Transformation
We wish to thank M. D. Tuduhl, Sr., for assistance in the of the Stroma of Mammary Carcinomas that Stimulated
photography. Fibroblastic Growth in vitro. Cancer Res., 5: 257-264, 1945.
20. Maeir, D. M., and Zaiman, II. The Development of Lysosomes
REFERENCES in Rat Skeletal Muscle in Trichinous Myositis. J. Histochem.
1. Abercrombie, M., and Ambrose, K. I. The Surface Properties Cytochem., 1J,:390-400, 1900.
of Cancer Cells: A Review. Cancer Res., 32: 525-548, 1902. 21. Noi, V. J. Quoted by J. M. Vasiliev and V. I. Guelstein. In:
2. Abercrombie, M., Heaysman, J. E. M., and Karthauser, H. M. Local Cell Interactions in Neoplasms and in the Foci of Car-
Social Behavior of Cells in Tissue Culture. III. Mutual Influ cinogenesis. Progr. Exptl. Tumor Res., 8: 26-65, 1906.
ence of Sarcoma Cells and Fibroblasta, Exptl. Cell Res., 13: 22. Pearse, A. G. E., Hess, R., and Snyder, R. Stromal and Tumor
276-291, 1957. Cathepsins and Their Relationships to Metabolic Activity
3. Ambrose, E. J., Easty, D. M., and Jones, P. C. T. Specific and Invasiveness. In: M. J. Brennan and W. L. Simpson (eds.),
Reactions of Poly-electrolytes with the Surfaces of Normal Biological Interactions in Normal and Neoplastic Growth;
and Tumor Cells. Brit. J. Cancer, 12: 439-447, 1958. Henry Ford Hospital International Symposium, pp. 057-670.
4. Argyris, B. F., and Argyris, T. S. Mechanism of Mammary Boston- Little Brown and Co., 1962.
Duct Stimulation by Tumor Transplants. Cancer Res., 21: 23. Ranadive, K. J., and Bhide, S. V. Tissue Interactions Between
75-81, 1961. Normal and Malignant Cells In: M. J. Brennan and W. L.
5. Bennet, L. R., and Cannon, F. E. Effects of Lytic Agents on Simpson (eds.), Biological Interactions in Normal and Neo
Plasma Membrane of Ehrlich Ascites Tumor Cells and Mouse plastic Growth; Henry Ford Hospital International Sympo-
Erythrocytes. J. Nati. Cancer Inst., 19: 999-1012, 1957. posium, pp. 337-354. Boston: Little Brown and Co., 1902.
6. Berenblum, I. The Nature of Tumor Growth. In: II. Florey 24. Sabatini, D. D., Bensch, K., and Barrnet, R. J. Cytochemistry
(ed.), General Pathology, Ed. 3, pp. 528-550. Philadelphia: and Electron Microscopy. The Preservation of Cellular Ultra-
W. B. Saunders Co., 1962. structure and Enzymatic Activity by Aldehyde Fixation. J.
7. Brandes, D., Anton, E., and Lam, K. W. Studies of L 1210 Cell Biol., 17: 19-58, 1903.
Leukemia. II. Ultrastructural and Cytochemical Changes after 25. Sylvcn, B. Ester Sulphuric Acids of High Molecular Weight
Treatment with Cyclophosphamide and Vitamin A. J. Nati. and Mast Cells in Mesenchymal Tumors. Acta Radio)., Suppl.
Cancer Inst., in press. 59, 1945.
2162 CANCER RESEARCH VOL. 27
Downloaded from cancerres.aacrjournals.org on January 17, 2021. © 1967 American Association for Cancer
Research.
Invasion of Muscle by L1210 Leukemia
20. Sylvén,B. The Biochemical Mechanism Underlying the De- 30 Vasiliev, J. M. The Role of Connective Tissue Proliferation in
structive Growth of Tumors. Acta UnióIntern. Contra Can- Invasive Growth of Normal and Malignant Tissues : A Review.
crum, 14: 61-02, 1958. Brit. J. Cancer, 18: 524-536, 1958.
27. Sylvén,B. The Host-Tumor Interzone and Tumor Invasion. 'Al. Vasiliev, J. M. The Local Stimulatory Effect of Normal Tissues
In: M. J. Brennan and W. L. Simpson (eds.), Biological Inter Upon the Growth of Tumor Cells. In: M. J. Brennan and W. L.
actions in Normal and Neoplastic Growth; Henry Ford Hos Simpson (eds.), Biological Interactions in Normal and Neo
pital International Symposium, pp. 635-655. Boston: Little plastic Growth; Henry Ford Hospital International Sym
Brown and Co., 1962. posium, pp. 299-309. Boston: Little Brown and Co., 19(i2.
28. Sylvén,B., and Malmgren, H. Topical Distribution of Pro- 32, Vasiliev, J. M., and Guelstein, V. I. Local Cell Interactions in
teolytic Activities in Some Transplanted Mice Tumors. Exptl. Neoplasms and in the Foci of Carcinogenesis. Progr. Exptl.
Cell Res., 8: 575-577, 1955. Tumor Res., 8: 26-65, 1966.
29. Sylvén, B., and Malmgren, II. The Histológica!Distribution of 33 Willis, R. A. The Direct Spread of Tumors. In: Pathology of
Proteinase and Peptidase Activity in Solid Tumor Trans Tumors, Ed. 2, pp. 147-165. London: Butter-worth and Co.
plants. Acta Radiol., Suppl. 154, 1957. (Publishers Ltd.), 1953.
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David Brandes, Elsa Anton, and Brian Schoficld
FIG. 1. Normal appearance of L1210 leukemic cells. The nuclei (.V) occupy a large portion of the cell. The cytoplasm is studded
with ribosomes and shows virus particles (V), some of which are of the intracisternal type (arrows). The Golgi complex ((?) is well
developed, hut the rough-surfaced endoplasmic reticulum (REIt) is scarce, a, X 10,500; 6, X 20,500; inset, X 30,000.
FIG. 2. Light micrograph. A positive reaction is seen in a few macrophages (arrows). The muscle fibers (asterisks) and the invading
leukemic cells show no apparent activity. Acid phosphatase stain, X 350.
FIG. 3. Electron micrograph. The leukemic cells (LK) contain few small lysosomes (Ly) which were not resolved in the light micro
graphs. Neither the muscle fiber (SK) nor the host-tumor (HT) interzone show any apparent activity. A', nuclei. Acid phosphatase
stain, X 11,500.
FIGS. 4 AND 5. One-micron sections from blocks embedded in Kpon from which ultrathin sections were then cut forelectron micros
copy. Toluidine blue stain.
FIG. 4. Invasion of smooth muscle of intestinal wall. Fragmented muscle fibers (arrows) are surrounded by leukemic cells. X 350.
FIG. 5. Invasion of skeletal muscle. Some fragmented fibers (arrows) are surrounded by leukemic cells, but others (asterisks) are seen
in areas showing no leukemic infiltration. X 350.
FIG. 6. Portion of a leukemic cell (LA') infiltrating a space between skeletal muscle fibers (SK). The plasma membrane (Pni) is still
intact and so is the sarcolemma (SI). The latter is externally coated by a thin layer of amorphous material with the appearance of a
basement membrane (Urn). The myofibrils (My) appear normal. N, nucleus; V, virus particles; Li, lipid droplets; Ct, connective tissue
fibers. X 20,000.
FIG. 7. Host-tumor interzone showing clusters of ribosomes (arrows) in the interstitial space. The plasma membrane of the leukemic
cell (LK) and the sarcolemma (SI) are still intact. JV, nucleus of leukemic cell; .V, nucleus of skeletal muscle fiber. X 10,500.
FIG. 8. Degenerating leukemic cell (/>K) in the midst of skeletal muscle fibers (SK). The nucleus (X) is undergoing pycnosis. The
plasma membrane is no longer visible, and the cytoplasm is in direct continuity with the interstitial space (arrows). The sarcolemma.
(Si) is intact and shows numerous pinocytotic vesicles. Virus particles are still present (V) in the tumor cell cytoplasm. X 21,500.
FIG. 9. Two necrobiotic leukemic cells (LK\, AA'2). Cytoplasmic debris have spread through the interstitial space (asterisk). An
adjacent muscle fiber (SK) appears normal and the sarcolemma (SI) is intact. Virus particles (V) are still seen in the necrobiotic leu
kemic cell. They are shown at higher magnification in the iipper right inset. Part of a macrophage (MP) is also visible. X 9,500.
Inset, X 18,500.
FIG. 10. Cytoplasmic materials from disintegrating leukemic cells are in continuity with the interstitial space. The nuclei (N) are
undergoing pycnosis. The skeletal muscle fiber (SK) contains numerous myelin bodies (My). X 18,500.
FIG. 11. A skeletal muscle fiber (SK) in contact with leukemic cell debris (asterisk) shows an area of hyperplastic marginal sarco-
plasm (star) containing abundant ribosomes and myelin bodies. X 20,000.
FIG. 12. Skeletal muscle fiber (SK) in an area of invasion. Hyperplastic marginal sarcoplasm containing an accumulation of hyper-
trophic mitochondria (M), abundant ribosomes, and a prominent Golgi complex (G). X 23,500.
FIG. 13. Area similar to that shown in Fig. 12 shows degenerative changes in mitochondria (arrows). A prominent Golgi complex (G)
is visible. SK, skeletal muscle fiber. X 21,500.
FIG. 14. Initial steps in the process of skeletal muscle disintegration. The sarcolemma is no longer visible and the surface of the
muscle fibers show a frayed appearance (arrow). A degenerating leukemic cell (LK) devoid of plasma membranes is present in this area.
X 11,500.
FIG. 15. The process of skeletal muscle degeneration is more advanced. The sarcolemma has disappeared, and the frayed appearance
of the surface is accentuated (arrows). The myofibrils start to disintegrate (asterisk). Abundant debris of tumor cell is seen in the inter
stitial space. X 23,500.
FIG. 16. Skeletal muscle fibers (SKi) in the process of fragmentation. The myofibrils are interrupted in an area of constriction (ar
rows). A second fiber (SKt) shows incipient fraying of the surface. No neoplastic cells are seen in this area, but the interstitial space
contains cell debris. N, nucleus of muscle cell. X 14,500.
FIG. 17. Isolated skeletal muscle fragments (SK) in an area which contains no tumor cells. Cellular debris are present in the inter
stitial spaces. N, nucleus of muscle cell. X 10,000.
FIG. 18. Diaphragmatic muscle in an area of ascites tumor cell invasion. The larger fragment (.4) contains mitochondria] aggregates
and shows striai ed fibrils (SF). Pinocytotic and finger-like projections at the cell surface are abundant. Small portions of muscle remain
attached to larger fragments by narrow Cytoplasmic bridges (arrows). No tumor cells are present in this area. X 10,000.
FIG. 19. Smooth muscle (SM) from intestinal wall, in the neighborhood of an area of leukemic infiltration. Aggregates of mitochondria
(M), hypertrophie Golgi elements (G), and increased pinocytotic activity are observed in many muscle cells. X 20,000.
FIG. 20. Smooth muscle fibers (SM) surrounded by leukemic cells (LK). Abundant rosettes of ribosomes are free in the interstitial
spaces (asterisk) between both cell types. Mitochondria, some with altered cristae, Golgi elements (G), myelin bodies (arrows), and ribo
somes are concentrated near such areas. X 21,500.
FIG. 21. Altered leukemic cell (AA'i) with marked dilation of the perinuclear cisterna (Pc) and swollen mitochondria (M). The plasma
membrane has almost completely disappeared, and areas of fusion of tumor cell and muscle cells are frequent (asterisk). Mitochondrial
aggregates are seen in such areas. The plasma membrane (Pm) of another leukemic cell (LA%) is intact. X 30,500.
FIG. 22. Extensive fusion of altered leukemic cell (LK) and smooth muscle (SM). Morphologic alterations in leukemic cell mitochon
dria (asterisk) are more obvious. Tumor cell ribosomes are in contact or within muscle cytoplasm which also shows vacuoles (Va) con
taining ribosomes. Initial lytic changes are seen in the muscle fiber at the area of the fusion with the leukemic cells (arrow). X 21,500.
FIG. 23. a, a mitochondrion (M) is partially within a leukemic cell (LK) and partially within a smooth muscle filier (SM). b, Arrow
shows where sarcolemma (SI) becomes interrupted. Muscle fibrils (star) have also penetrated the leukemic cell cytoplasm, a, X 10,500;
b, X 18,500.
FIG. 24. Capillary in area containing tumor cell debris. The endothelium is hypertrophie and shows enhanced pinocytotic activity
(arrous). X 11,500.
FIG. 25. Hyperplasia of Golgi elements (G) and of rough endoplasmic reticulum in capillary endolhelium in the area of tumor cell
invasion. X 18,500.
2164 CANCER RESEARCH VOL. 27
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Research.Invasion of Skeletal and Smooth Muscle by L1210 Leukemia
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Cancer Res 1967;27:2159-2178.
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