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Ultrastructure of the Dermal-epidermal Junction in Experimentally Induced Tumors and Human Oral Lesions*
Taa JoURNAL OF IrevEsTioATIvE DERMATOLOGY
Copyright
Vol. 52, No. 8
Printed in U.S.A.
1559 by The Williams & Wilkins Co.
ULTRASTRUCTURE OF THE DERMAL-EPIDERMAL JUNCTION IN EXPERIMENTALLY INDUCED TUMORS AND HUMAN ORAL LESIONS* D. A. WOODS, B.Sc., PH.D.t AND C. J. SMITH, PH.D., B.D.S., LD.S.R.C.S4
Electron microscopic observations have revealed intradermal protrusions of epithelial basal cells in tumors of the human uterine cervix (1, 2, 3, 4), human breast carcinoma
in order to obtain a full range of types and size gradations. Untreated pouches and pouches in which DMBA painting had been discontinued for
ferent animal species (5, 6, 7) and human
cally normal mucosa overlying unerupted impacted teeth prior to their surgical removal.
three months, were also sampled.
Biopsies of hyperkeratotie human oral mucosa (Hallowes, unpublished results), experimental were obtained for electron microscopy and routine tumors induced by various carcinogens in dif- histology. Specimens were taken also from clinisenile keratoma and laryngeal papilloma (5).
Encroachment of basal cells into the dermis has also been demonstrated in guinea-pig skin during treatment with hydroxyanisoles, chemicals of no known earcinogenicity (9). Similar morphological changes occur in guinea-pig skin
Tissues were fixed in 2% phosphate buffered osmium tetroxide for from one to three hours and embedded in Araldite epoxy resin. Sections were stained with methanolie uranyl acetate (12) and aqueous lead citrate (13). RESULTS
after intraeutaneous injection of Clostridial Hamster Carcino genesis
eollagenase (10).
Breakdown of tissue organization at the der-
There was a minimum latent period of nine mal-epidermal junction has been reported as weeks from the commencement of painting, occurring in essentially 'normal', rapidly grow- during which an initial inflammatory reaction ing tissues, e.g. the growing hair follicle (7) occurred and slowly subsided, and epidermal and the regenerating newt limb (11). hyperplasia developed (14). During this latent The present paper is a preliminary report on period electron microscopy revealed some disepidermal pseudopodial invasion of the dermis organization of the eollagenous dermis, often in chemically induced hamster cheek pouch accompanied by an increase in the dermal cell tumors, and similar phenomena observed in population. Passage of inflammatory cells from human hyperkeratotie oral mueosa.
the dermis into the epidermis caused extensive disruption of the basal lamina; migrating cells
METHODS
were sometimes found lying between the
Cheek pouches of Golden Syrian hamsters lamina densa and the plasma membranes of (Mesocricetus ourolus) were painted thrice the basal cells. During the recovery from inweekly with 0.5% 7,12-dimethylbenz(a)anthracene
flammation, new material was apparently elaborated to cover the exposed surface of epidermal after three, six and nine weeks of DMBA treat- cells which had become freed from the lamina ment. Polypoid tumours that began to develop densa, causing duplication of the basal lamina. thereafter were harvested at random intervals After nine weeks of DMBA treatment small spontaneous breaks were noted in the lamina Received July 12, 1968; accepted for publication August 26, 1968. densa, and basal cells of the hyperplastie epiThe authors wish to thank Professor B. Cohen of the Department of Dental Science, Royal Col- dermis occasionally extended short pleomorphic lege of Surgeons, for his advice and Mr. M. pseudopodia through the basal lamina into the Awty of the Dental Department, The Queen Vic- dermis. Smoll tumors also began to occur from toria Hospital, East Grinstead, Sussex, for pro(DMBA) in liquid paraffin. Specimens for electron microscopy were taken from separate pouches
this time on; these displayed market epider-
viding the specimens of human tissue. * From the Electron Microscopy Unit, Imperial
mal cell hyperactivity accompanied by a rapid Cancer Research Fund, London, W.C2.t and the increase in the incidence of basal cell pseudoDepartment of Dental Science, Royal College of podia (Fig. 1). The increase in size and number Surgeons. London, W.C2. t Prophit Cancer Research Student, Depart- of these intradermal extrusions appeared to be ment of Dental Science, Royal College of Sur- in proportion to the disintegration of the eons, London, \\TC2
259
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
260
--S
Fm. 1. Premalignant hamster tumor; basal cell pseudopodia (P) growing through the lamina densa (1) and invading the dermis (D).
superficial dermis; the process culminated in pseudopodia was still in evidence. In regions almost total lysis of the connective tissue which lacked basal lamina, the plasma memand repopulation of dermal regions by epi- branes of the epidermal cells adjacent to the dermal pseudopodia (Fig. 2). It is possible that connective tissue were extremely convoluted some of the pseudopodia lose the connection but apparently lacked true pseudopodia. Pseuwith their cell of origin and become isolated dopodial infiltration was rarely found in the vesicles in the dermis.
dermis of these tumors.
TUmors developed, after an extended latent
The epidermal extrusions had many of the period, in cheek pouch mueosa that had re- morphological attributes normally associated
ceived applications of DMBA for three months with pseudopodia observed in connection with and had then been left unpainted for a further phagocytosis or ameboid movement. They were three months. These tumors also contained bordered by a plasma membrane which lacked massive dermal infiltrations of pseudopodia. attachment complexes, and contained a homogAll the tumors, if allowed to develop, showed eneous ectoplasm devoid of organdIes. At the histological features characteristic of invasive junction with the cell proper, the terminal web squamous cell carcinomas. was frequently very prominent.
Large tumors in advanced stages of maligHuman Biopsies nancy had extensive dermal disruption and often total absence of basal lamina. Where traces Although some of these specimens were obof the lamina densa remained, the formation of tained from clinically normal tissue most of
ULTRASTRUCTURE OF EXPERIMENTAL TUMORS AND HUMAN ORAL LESIONS
Fie. 2. Premalignant hamster tumor with advanced lysis of the connective tissue.
FIG. 3. Dermal-epidermal junction of human hyperkeratotic oral mucosa.
261
262
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
them showed a mild chronic inflammatory cell at weak points in the basal lamina but that infiltration in the dermal component. Duplica- they are active appendages of the cell surface. Several instances of pseudopodium formation tion of the lamina densa occurred in apparently healthy oral mucosa to a limited extent but associated with the development of carcinomas was more marked in hyperkeratotic tissue. have already been cited (see above). Similar The fibrillar complexes on the dermal face of phenomena have been seen in human oral the lamina densa (15) indicated that this was a mucosa of a hyperkeratotic nature and postrue multi-lamination and not an artefact due sible malignant potential. The significance of these observations remains to be evaluated. to folding of the tissue. The dermis of the pathological tissues was Absence of pseudopodia from the epidermis often heavily populated with macrophages and of clinically normal hut histologically inflamed numerous mast cells in various stages of de- human oral mucosa indicates that the effect is granulation. Dermal cells were frequently close probably not due to inflammatory changes. to the basal lamina. Breaks were evident in Similarly in the DMBA treated hamster cheek the lamina debsa, opposite basal cell junctions, pouch, pseudopodia are not seen until the inwhich could he attributed to the passage of in- flammatory reaction has subsided and hyperflammatory cells through the basal layer of the plasia is established. It is possible that the epidermis. In addition, however, small epi- pseudopodia are associated with the hyperplastic dermal protrusions occasionally were found state and while there is no evidence that they penetrating the lamina densa where no inflamma- are directly involved in the etiology of any
tory activity was in evidence. A characteristic ensuing malignancy the possibility cannot be region from one biopsy is illustrated in Figure 3.
discounted.
This shows dissolution of the collagen fibres, interruptions in the lamina densa and a pseudopodium of similar morphology to those observed in the hyperplastic epidermis and early polyps of carcinogen treated hamster cheek pouch. Histological examination of the lesion
Disturbance of the collagcnous dermis is a feature common to both the hamster tumors
tism of some basal cell nuclei and individual cell
(10) causes ultrastructural changes at the
and the human lesions. In the former, complete
destruction of the sub-epidermal connective tissue may occur which suggests the involvement of a proteolytic enzyme. In guinea-pig showed loss of basal cell polarity, hyperchroma- skin, intracutaneous injection of collagenase dyskeratosis, features which were considered dermal-epidermal junction similar to phenomena associated with premalignant tissues. The inindicative of impending malignancy. jected collagenase has been shown to destroy DJscussroN the lamina densa even more rapidly than the Chronic inflammation of the mucosa is often sub-epidermal collagen, a finding which may be accompanied by damage to the basal lamina. correlated with the progressive breakdown of
Exposure of the plasma membrane of basal cells to the connective tissue may stimulate the formation of a subsidiary lamina densa, leaving the remnants of the primary basement membrane free in the dermis. A fundamental feature of epidermal carcinogenesis appears to be the failure of the neoplastic epithelium to maintain a basal lamina at its
the lamina densa that accompanies dermal lysis in experimental tumors.
The formation of epidermal pseudopodia has been observed following applications of hydroxy-
anisole to guinea pig ear epidermis (9). Under
the experimental conditioos employed, the effects of this chemical were reversed upon cessation of treatment; the effects of DMBA interface with the underlying connective tissue. in hamster cheek pouch, however, escalated This malfunction is exemplified at focal re- irrevocably and were not entirely dependent gions of hyperactivity of the plasma membrane, upon continued application of DMBA. Further
the pseudopodia, and culminates in total loss investigations are being undertaken to ascerof lamina densa in advanced carcinomas. The tain the influence of hydroxyanisole on the morphology of the pseudopodia i.e. the absence hamster cheek pouch mucosa.
of organellcs and the demarcation between It is probable that the maintenance of muectoplasm and endoplasm, suggests that they cosal normality is a product of dcrmal-epiare not merely passive extrusions of cytoplasm dermal interaction. Treatment with chemicals
ULTRASTRUCTURE OF EXPERIMENTAL TUMORS AND HUMAN ORAL LESIONS
such as DMBA or hydroxyanisole (9) apparently disturbs the balance of this interaction and causes sub-microscopic invasion of the connective tissue by epidermal pseudopodia. Such chemical interference by polycyclic hydrocarbons of the DMBA type may be a function of cytotoxicity quite divorced from their ability to
potentiate malignancy. Although no link has
263
of normal epithelium, carcinoma in situ and invasive carcinoma of uterine cervix utilising
electron microscopy and bistoehemical
methods. Aeta Cytol., 5: 369, 1961. 3. Hinglais-Guillaud, N., Moricard, R. and Bernhard, W.: Ultrastruetures des cancers pavi-
menteux invasifs du col utdrin ches la femme. Bull. Ass. franc. Cancer, 48: 1961.
283,
4. Song, J.: The Human Uterus. Charles C Thomas, Springfield, Illincis, 1964.
been established between this sub-microscopic lesion and carcinogenesis, several cases have
5. Frei, J. V.: The fine structure of the basement membrane in epidermal tumors. J. Cell Biol.,
been documented, both experimental and
6. Fasske, E. and Morgenroth, K.: Electron mi-
spontaneous in origin, in which this lesion has been shown to be associated with impending
basement membrane to the squamous cell carcinomas in animals. Oacnlogia, 20: 113,
15: 335, 1962.
croscopic studies on the relation of the
1966. 7. Tarin, D.:
malignancy. STJMMART
Hamster cheek pouch mucosa was treated with the carcinogen 7, 12-dimethylbenz (a) an-
Sequential electron microscopical study of experimental mouse skin careino-
genesis. Tnt. J. Cancer, 2: 195, 1967.
8. Sugir, J.: An electron microscopic study of early invasive growth in human skin tumours and laryngeal carcinoma. Europ. J. Cancer, 4: 33, 1968.
thracene and biopsies were taken at regular 9. Seal, P., Riley, P. A. and Inman, P. R.: Basal intervals for examination in the electron microcell encroachment into the dermis caused by applications of hydroxyanisole. J. Invest. scope. After a latent period, the epidermis beDerm., 52: 264, 1969. came hyperplastie and discontinuities occurred 10. Kabl, F. R. and Pearson, R. W.: Ultrastructural studies of experimental vesiculation. in the lamina densa. Pseudopodia extended from TI. Collagenase. J. Invest. Derm., 49: 616, the basal epithelial cells into the connective 1967. tissue, at first an isolated phenomenon but be- 11. Salpeter, M. M. and Singer, M.: Differentiation of the submicroscopic adepidermal coming widespread in developing tumors. Dismembrane during limb regeneration in adult solution of the sub-epidermal connective tissue Triturus, including a note on the use of the term basement membrane. Anat. Ree. 786: and the proliferation of pseudopodia appeared 27, 1960. to be correlated. Similar dermal-epidermal tissue 12. Stempak, J. G. and Ward, R. T.: An improved interaction was seen in human hyperkeratotie staining method for electron microscopy. J. Cell Biol., 92: 697, 1964. oral mneosa. 13. Reynolds, E. S.: The use of lead citrate at high pH as an electron-opaque stain in elecREFERENCES tron microscopy. J. Cell Biol., 17, 208, 1963. 14. Camilleri, G. E. and Smith, C. J.: Exfoliative 1. Luibel, F. J., Sanders, E. and Ashworth, C. T.: An electron microscopic study of carcinoma in situ and invasive carcinoma of the cervix uteri. Cancer Res., 20: 357, 1960. 2. Ashworth, C. T., Stembridge, V. A. and Luibel, F. J.: A study of basement membranes
cytology in experimental oral carcinogenesis. Acta Cytnl., 8: 85, 1964.
15. Susi, F. R., Belt, W. D. and Kelly, J. W.: Fine structure of fibrillar complexes associated
with the basement membrane in human oral mucosa. J. Cell Binl., 84: 686, 1967.
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
94
linolenic acid extract. Arch. This pdf is a scanned copy UV of irradiated a printed document.
24. Wynn, C. H. and Iqbal, M.: Isolation of rat
skin lysosomes and a comparison with liver Path., 80: 91, 1965. and spleen lysosomes. Biochem. J., 98: lOP, 37. Nicolaides, N.: Lipids, membranes, and the 1966.
human epidermis, p. 511, The Epidermis
Eds., Montagna, W. and Lobitz, W. C. Acascopic localization of acid phosphatase in demic Press, New York. human epidermis. J. Invest. Derm., 46: 431, 38. Wills, E. D. and Wilkinson, A. E.: Release of 1966. enzymes from lysosomes by irradiation and 26. Rowden, C.: Ultrastructural studies of kerathe relation of lipid peroxide formation to tinized epithelia of the mouse. I. Combined enzyme release. Biochem. J., 99: 657, 1966. electron microscope and cytochemical study 39. Lane, N. I. and Novikoff, A. B.: Effects of of lysosomes in mouse epidermis and esoarginine deprivation, ultraviolet radiation and X-radiation on cultured KB cells. J. phageal epithelium. J. Invest. Derm., 49: 181, 25. Olson, R. L. and Nordquist, R. E.: Ultramicro-
No warranty is given about the accuracy of the copy.
Users should refer to the original published dermal cells. Nature, 216: 1031, 1967. version of1965. the material. vest. Derm., 45: 448, 28. Hall, J. H., Smith, J. G., Jr. and Burnett, S. 41. Daniels, F., Jr. and Johnson, B. E.: In prepa1967.
Cell Biol., 27: 603, 1965.
27. Prose, P. H., Sedlis, E. and Bigelow, M.: The 40. Fukuyama, K., Epstein, W. L. and Epstein, demonstration of lysosomes in the diseased J. H.: Effect of ultraviolet light on RNA skin of infants with infantile eczema. J. Inand protein synthesis in differentiated epi-
C.: The lysosome in contact dermatitis: A ration. histochemical study. J. Invest. Derm., 49: 42. Ito, M.: Histochemical investigations of Unna's oxygen and reduction areas by means of 590, 1967. 29. Pearse, A. C. E.: p. 882, Histochemistry Theoultraviolet irradiation, Studies on Melanin, retical and Applied, 2nd ed., Churchill, London, 1960.
30. Pearse, A. C. E.: p. 910, Histacheini.stry Thearetscal and Applied, 2nd ed., Churchill, London, 1960.
31. Daniels, F., Jr., Brophy, D. and Lobitz, W. C.: Histochemical responses of human skin fol-
lowing ultraviolet irradiation. J. Invest. Derm.,37: 351, 1961.
32. Bitensky, L.: The demonstration of lysosomes by the controlled temperature freezing section method. Quart. J. Micr. Sci., 103: 205, 1952.
33. Diengdoh, J. V.: The demonstration of lysosomes in mouse skin. Quart. J. Micr. Sci., 105: 73, 1964.
34. Jarret, A., Spearman, R. I. C. and Hardy, J. A.:
Tohoku, J. Exp. Med., 65: Supplement V, 10, 1957.
43. Bitcnsky, L.: Lysosomes in normal and pathological cells, pp. 362—375, Lysasames Eds., de Reuck, A. V. S. and Cameron, M. Churchill, London, 1953.
44. Janoff, A. and Zweifach, B. W.: Production of inflammatory changes in the microcirculation by cationic proteins extracted from lysosomes. J. Exp. Med., 120: 747, 1964.
45. Herion, J. C., Spitznagel, J. K., Walker, R. I. and Zeya, H. I.: Pyrogenicity of granulocyte lysosomes. Amer. J. Physiol., 211: 693, 1966.
46. Baden, H. P. and Pearlman, C.: The effect of ultraviolet light on protein and nucleic acid synthesis in the epidermis. J. Invest. Derm.,
Histochemistry of keratinization. Brit. J. 43: 71, 1964. Derm., 71: 277, 1959. 35. De Duve, C. and Wattiaux, R.: Functions of 47. Bullough, W. S. and Laurence, E. B.: Mitotic control by internal secretion: the role of lysosomes. Ann. Rev. Physiol., 28: 435, 1966. the chalone-adrenalin complex. Exp. Cell. 36. Waravdekar, V. S., Saclaw, L. D., Jones, W. A. and Kuhns, J. C.: Skin changes induced by
Res., 33: 176, 1964.
Copyright
Vol. 52, No. 8
Printed in U.S.A.
1559 by The Williams & Wilkins Co.
ULTRASTRUCTURE OF THE DERMAL-EPIDERMAL JUNCTION IN EXPERIMENTALLY INDUCED TUMORS AND HUMAN ORAL LESIONS* D. A. WOODS, B.Sc., PH.D.t AND C. J. SMITH, PH.D., B.D.S., LD.S.R.C.S4
Electron microscopic observations have revealed intradermal protrusions of epithelial basal cells in tumors of the human uterine cervix (1, 2, 3, 4), human breast carcinoma
in order to obtain a full range of types and size gradations. Untreated pouches and pouches in which DMBA painting had been discontinued for
ferent animal species (5, 6, 7) and human
cally normal mucosa overlying unerupted impacted teeth prior to their surgical removal.
three months, were also sampled.
Biopsies of hyperkeratotie human oral mucosa (Hallowes, unpublished results), experimental were obtained for electron microscopy and routine tumors induced by various carcinogens in dif- histology. Specimens were taken also from clinisenile keratoma and laryngeal papilloma (5).
Encroachment of basal cells into the dermis has also been demonstrated in guinea-pig skin during treatment with hydroxyanisoles, chemicals of no known earcinogenicity (9). Similar morphological changes occur in guinea-pig skin
Tissues were fixed in 2% phosphate buffered osmium tetroxide for from one to three hours and embedded in Araldite epoxy resin. Sections were stained with methanolie uranyl acetate (12) and aqueous lead citrate (13). RESULTS
after intraeutaneous injection of Clostridial Hamster Carcino genesis
eollagenase (10).
Breakdown of tissue organization at the der-
There was a minimum latent period of nine mal-epidermal junction has been reported as weeks from the commencement of painting, occurring in essentially 'normal', rapidly grow- during which an initial inflammatory reaction ing tissues, e.g. the growing hair follicle (7) occurred and slowly subsided, and epidermal and the regenerating newt limb (11). hyperplasia developed (14). During this latent The present paper is a preliminary report on period electron microscopy revealed some disepidermal pseudopodial invasion of the dermis organization of the eollagenous dermis, often in chemically induced hamster cheek pouch accompanied by an increase in the dermal cell tumors, and similar phenomena observed in population. Passage of inflammatory cells from human hyperkeratotie oral mueosa.
the dermis into the epidermis caused extensive disruption of the basal lamina; migrating cells
METHODS
were sometimes found lying between the
Cheek pouches of Golden Syrian hamsters lamina densa and the plasma membranes of (Mesocricetus ourolus) were painted thrice the basal cells. During the recovery from inweekly with 0.5% 7,12-dimethylbenz(a)anthracene
flammation, new material was apparently elaborated to cover the exposed surface of epidermal after three, six and nine weeks of DMBA treat- cells which had become freed from the lamina ment. Polypoid tumours that began to develop densa, causing duplication of the basal lamina. thereafter were harvested at random intervals After nine weeks of DMBA treatment small spontaneous breaks were noted in the lamina Received July 12, 1968; accepted for publication August 26, 1968. densa, and basal cells of the hyperplastie epiThe authors wish to thank Professor B. Cohen of the Department of Dental Science, Royal Col- dermis occasionally extended short pleomorphic lege of Surgeons, for his advice and Mr. M. pseudopodia through the basal lamina into the Awty of the Dental Department, The Queen Vic- dermis. Smoll tumors also began to occur from toria Hospital, East Grinstead, Sussex, for pro(DMBA) in liquid paraffin. Specimens for electron microscopy were taken from separate pouches
this time on; these displayed market epider-
viding the specimens of human tissue. * From the Electron Microscopy Unit, Imperial
mal cell hyperactivity accompanied by a rapid Cancer Research Fund, London, W.C2.t and the increase in the incidence of basal cell pseudoDepartment of Dental Science, Royal College of podia (Fig. 1). The increase in size and number Surgeons. London, W.C2. t Prophit Cancer Research Student, Depart- of these intradermal extrusions appeared to be ment of Dental Science, Royal College of Sur- in proportion to the disintegration of the eons, London, \\TC2
259
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
260
--S
Fm. 1. Premalignant hamster tumor; basal cell pseudopodia (P) growing through the lamina densa (1) and invading the dermis (D).
superficial dermis; the process culminated in pseudopodia was still in evidence. In regions almost total lysis of the connective tissue which lacked basal lamina, the plasma memand repopulation of dermal regions by epi- branes of the epidermal cells adjacent to the dermal pseudopodia (Fig. 2). It is possible that connective tissue were extremely convoluted some of the pseudopodia lose the connection but apparently lacked true pseudopodia. Pseuwith their cell of origin and become isolated dopodial infiltration was rarely found in the vesicles in the dermis.
dermis of these tumors.
TUmors developed, after an extended latent
The epidermal extrusions had many of the period, in cheek pouch mueosa that had re- morphological attributes normally associated
ceived applications of DMBA for three months with pseudopodia observed in connection with and had then been left unpainted for a further phagocytosis or ameboid movement. They were three months. These tumors also contained bordered by a plasma membrane which lacked massive dermal infiltrations of pseudopodia. attachment complexes, and contained a homogAll the tumors, if allowed to develop, showed eneous ectoplasm devoid of organdIes. At the histological features characteristic of invasive junction with the cell proper, the terminal web squamous cell carcinomas. was frequently very prominent.
Large tumors in advanced stages of maligHuman Biopsies nancy had extensive dermal disruption and often total absence of basal lamina. Where traces Although some of these specimens were obof the lamina densa remained, the formation of tained from clinically normal tissue most of
ULTRASTRUCTURE OF EXPERIMENTAL TUMORS AND HUMAN ORAL LESIONS
Fie. 2. Premalignant hamster tumor with advanced lysis of the connective tissue.
FIG. 3. Dermal-epidermal junction of human hyperkeratotic oral mucosa.
261
262
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
them showed a mild chronic inflammatory cell at weak points in the basal lamina but that infiltration in the dermal component. Duplica- they are active appendages of the cell surface. Several instances of pseudopodium formation tion of the lamina densa occurred in apparently healthy oral mucosa to a limited extent but associated with the development of carcinomas was more marked in hyperkeratotic tissue. have already been cited (see above). Similar The fibrillar complexes on the dermal face of phenomena have been seen in human oral the lamina densa (15) indicated that this was a mucosa of a hyperkeratotic nature and postrue multi-lamination and not an artefact due sible malignant potential. The significance of these observations remains to be evaluated. to folding of the tissue. The dermis of the pathological tissues was Absence of pseudopodia from the epidermis often heavily populated with macrophages and of clinically normal hut histologically inflamed numerous mast cells in various stages of de- human oral mucosa indicates that the effect is granulation. Dermal cells were frequently close probably not due to inflammatory changes. to the basal lamina. Breaks were evident in Similarly in the DMBA treated hamster cheek the lamina debsa, opposite basal cell junctions, pouch, pseudopodia are not seen until the inwhich could he attributed to the passage of in- flammatory reaction has subsided and hyperflammatory cells through the basal layer of the plasia is established. It is possible that the epidermis. In addition, however, small epi- pseudopodia are associated with the hyperplastic dermal protrusions occasionally were found state and while there is no evidence that they penetrating the lamina densa where no inflamma- are directly involved in the etiology of any
tory activity was in evidence. A characteristic ensuing malignancy the possibility cannot be region from one biopsy is illustrated in Figure 3.
discounted.
This shows dissolution of the collagen fibres, interruptions in the lamina densa and a pseudopodium of similar morphology to those observed in the hyperplastic epidermis and early polyps of carcinogen treated hamster cheek pouch. Histological examination of the lesion
Disturbance of the collagcnous dermis is a feature common to both the hamster tumors
tism of some basal cell nuclei and individual cell
(10) causes ultrastructural changes at the
and the human lesions. In the former, complete
destruction of the sub-epidermal connective tissue may occur which suggests the involvement of a proteolytic enzyme. In guinea-pig showed loss of basal cell polarity, hyperchroma- skin, intracutaneous injection of collagenase dyskeratosis, features which were considered dermal-epidermal junction similar to phenomena associated with premalignant tissues. The inindicative of impending malignancy. jected collagenase has been shown to destroy DJscussroN the lamina densa even more rapidly than the Chronic inflammation of the mucosa is often sub-epidermal collagen, a finding which may be accompanied by damage to the basal lamina. correlated with the progressive breakdown of
Exposure of the plasma membrane of basal cells to the connective tissue may stimulate the formation of a subsidiary lamina densa, leaving the remnants of the primary basement membrane free in the dermis. A fundamental feature of epidermal carcinogenesis appears to be the failure of the neoplastic epithelium to maintain a basal lamina at its
the lamina densa that accompanies dermal lysis in experimental tumors.
The formation of epidermal pseudopodia has been observed following applications of hydroxy-
anisole to guinea pig ear epidermis (9). Under
the experimental conditioos employed, the effects of this chemical were reversed upon cessation of treatment; the effects of DMBA interface with the underlying connective tissue. in hamster cheek pouch, however, escalated This malfunction is exemplified at focal re- irrevocably and were not entirely dependent gions of hyperactivity of the plasma membrane, upon continued application of DMBA. Further
the pseudopodia, and culminates in total loss investigations are being undertaken to ascerof lamina densa in advanced carcinomas. The tain the influence of hydroxyanisole on the morphology of the pseudopodia i.e. the absence hamster cheek pouch mucosa.
of organellcs and the demarcation between It is probable that the maintenance of muectoplasm and endoplasm, suggests that they cosal normality is a product of dcrmal-epiare not merely passive extrusions of cytoplasm dermal interaction. Treatment with chemicals
ULTRASTRUCTURE OF EXPERIMENTAL TUMORS AND HUMAN ORAL LESIONS
such as DMBA or hydroxyanisole (9) apparently disturbs the balance of this interaction and causes sub-microscopic invasion of the connective tissue by epidermal pseudopodia. Such chemical interference by polycyclic hydrocarbons of the DMBA type may be a function of cytotoxicity quite divorced from their ability to
potentiate malignancy. Although no link has
263
of normal epithelium, carcinoma in situ and invasive carcinoma of uterine cervix utilising
electron microscopy and bistoehemical
methods. Aeta Cytol., 5: 369, 1961. 3. Hinglais-Guillaud, N., Moricard, R. and Bernhard, W.: Ultrastruetures des cancers pavi-
menteux invasifs du col utdrin ches la femme. Bull. Ass. franc. Cancer, 48: 1961.
283,
4. Song, J.: The Human Uterus. Charles C Thomas, Springfield, Illincis, 1964.
been established between this sub-microscopic lesion and carcinogenesis, several cases have
5. Frei, J. V.: The fine structure of the basement membrane in epidermal tumors. J. Cell Biol.,
been documented, both experimental and
6. Fasske, E. and Morgenroth, K.: Electron mi-
spontaneous in origin, in which this lesion has been shown to be associated with impending
basement membrane to the squamous cell carcinomas in animals. Oacnlogia, 20: 113,
15: 335, 1962.
croscopic studies on the relation of the
1966. 7. Tarin, D.:
malignancy. STJMMART
Hamster cheek pouch mucosa was treated with the carcinogen 7, 12-dimethylbenz (a) an-
Sequential electron microscopical study of experimental mouse skin careino-
genesis. Tnt. J. Cancer, 2: 195, 1967.
8. Sugir, J.: An electron microscopic study of early invasive growth in human skin tumours and laryngeal carcinoma. Europ. J. Cancer, 4: 33, 1968.
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