Modulation by vitamin C of tumour incidence and inhibition in oral carcinogenesis.

This study reports the mechanism of involvement of vitamin C in the pathogenesis of induced oral carcinogenesis in the hamster cheek pouch epithelium. This site was exposed to either DMBA singly or in combination with vitamin C or DMBA for 7 weeks followed by only vitamin C till tumour induction. Macroscopically, vitamin C reduces the epithelial tumour incidence in the hamster cheek pouch. Microscopically tumours induced by DMBA alone were well differentiated squamous carcinomas in all the animals, whereas those induced by DMBA plus vitamin C were papillary epidermoid carcinomas with minimal invasion. These observations suggest that vitamin C is capable of restricting the growth of the initiated cells and does not allow the invasion of the subepithelium. The mechanism of action of vitamin C in the process of restricting growth and invagination was clearly shown in ultrastructural pathology too. In vitamin C exposed tumours the dermal epidermal junction of the cheek pouch epithelium maintains basement membrane integrity, with the presence of cellular organelles and cytoskeletal structure of the cells.

Periodic histopathological and ultrastructural changes of excess vitamin A on oral carcinogenesis.

In this study initially a precancerous condition, leukoplakia, was develop at 6 weeks treatment of DMBA whereas in the animals treated both DMBA + Vit. A, leukoplakia was seen at 10 weeks followed by papilloma or nodules at 12 weeks. Tumours induced by DMBA were more in number than DMBA + Vit. A treated tumours. The histological and ultrastructural changes were enhanced and prominent in DMBA treated animals at 12 weeks, where as these changes were considerably less in animals treated with DMBA + vit. A at 12 weeks.

Light and electron microscopic study of hamster cheek pouch treated with 9,10 dimethyl 1,2 benzanthracene and retinoic acid.

The present study reports light and electron microscopic observations of hamster cheek pouch epithelium exposed to 25 micrograms DMBA (DMBA = 9,10 Dimethyl, 1,2 Benz(a)-nthracene, RA = Retinoic Acid) and 25 micrograms DMBA along with 25 micrograms, 50 micrograms and 100 micrograms retinoic acid. Significant delay in tumour induction was observed in the animals treated with DMBA + retinoic acid. DMBA + retinoic acid treated cheek pouch developed papillary epidermoid carcinomas which were less invasive and less keratinized than only DMBA treated animals. At cellular level DMBA treated animals showed keratinized cells with thick bundles of tonofilaments, broken basement membrane, wide intercellular spaces and loss of desmosomal attachments, whereas animals treated with DMBA + retinoic acid showed decrease in the intercellular spaces, maintenance of basement membrane and intracellular organelles with suppression of keratinization, indicating the differentiating effect of retinoic acid on DMBA transformed cells of hamster cheek pouch.

Differential effects of phorbol ester tumor promoters on 3-methylcholanthrene-induced epithelial and mesenchymal skin tumorigenesis.

The effects of TPA, PDD, PDB, PDA, or MEZ on epithelial and mesenchymal skin tumors induced by a s.c. injection of MCA were studied histologically. Group-I mice received only MCA. At 6 weeks after MCA injection, mice in groups II to VII received acetone, 1.8 nmol TPA, PDD, PDB, PDA, or 6.1 nmol MEZ respectively in 0.1 ml acetone twice weekly until tumor development. Alterations in skin tumor induction patterns were also studied in animals that had been exposed to TPA or acetone for 10 weeks prior to s.c. injection of MCA. Exposure of mouse skin to TPA before or after carcinogen administration increased 2- to 3.5-fold, the incidence of carcinoma and mixed tumors of epithelial and mesenchymal histogenesis. The average time of tumor induction decreased in mice treated with MCA + TPA and 100% of the test animals in the TPA + MCA group developed tumors. In contrast, TPA-related phorbol esters inhibited skin tumor development, particularly trichoepithelioma and fibrosarcoma and increased the average time of tumor induction.

Ultrastructural analysis of epidermal hyperplasia induced by multiple 12-0-tetradecanoyl-phorbol-13-acetate (TPA) treatment of mouse skin.

Stationary epidermal hyperplasia induced by exposure of mouse skin to 1-6 TPA applications was analyzed by electron microscopy and found to be of two types. Intermingled orderly and irregularly stratified hyperplastic regions observed prominently after a single TPA application gave way, on multiple treatment, to epidermal hyperplasia populated by either cuboidal cells with expanded cytoplasm or by highly polar, narrow, tall and pleomorphic cells. Both cell types were poorly differentiated and displayed a paucity of intact desmosomal junctions, resulting in an incohesive tissue structure in which a number of phenotypic variants were expressed. The variants were markedly less mature than the adjacent cells and showed basal cell phenotype, acquisition of secretory activity or a disturbed mitotic process, resulting in the formation of binucleated cells. The observations suggest that the disturbed mitotic process, poor cellular differentiation and induction of metaplasia could be the mode by which an initiated cell may express its tumor phenotype and escape differentiation during the early stage of TPA promotion.

Ultrastructural localization of 5'nucleotidase in human normal and malignant lymphoid cells.

5'Nucleotidase (EC 3.1.3.5), a purine pathway enzyme, occurs as a cellular ectoenzyme. Widely differing 5'N activity has been reported in different types of lymphoid cells and appears to be related to lymphocyte function, differentiation and immunological subtype. This paper reports a study on the ultrastructural distribution of 5'N in in different types of lymphoid leukemias and non-Hodgkin's lymphomas. In lymphoid leukemias, only cALL cells showed strong 5'N staining. In CLL, PLL and HCL the intensity of staining was less than in normal cells. In the NHL group, the reaction pattern was mixed. Infiltrating neoplastic cells, specially the large lymphoid cells, consistently showed very mild activity of the enzyme, whereas follicular center cells and other mature lymphocytes were characterized by moderately strong to strong 5'N activity. These results support the view that 5'N is a marker of lymphocyte cell differentiation.

The relevance of gap junctions to stage I tumor promotion in mouse epidermis.

A previous paper reports that the potent tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA), has a time-dependent effect on mouse epidermal gap junctions. A single topical application of 1.0 micrograms TPA results in the absence of gap junctions from mouse interfollicular epidermis between 18 and 30 h post-treatment. This paper describes the dose-dependent effect of TPA on mouse epidermis. Observations indicate that only promoting doses of TPA affect the gap junctions. Similarly, while a low dose of the hyperplasiogenic compound mezerein (1.0 microgram) is ineffective, a higher dose (4.0 micrograms) results in a significant reduction in the gap junction number. One and two applications of TPA had identical effects. The potent inhibitor of both stage I and stage II of tumor promotion, Fluocinolone acetonide, used in combination with TPA, completely suppressed the hyperplasiogenic and the gap junction modulating effects of TPA. Retinoic acid, which inhibits only stage II of tumor promotion, did not influence the gap junction eliminating property of TPA. Tosylphenylalanine chloromethyl ketone which is a mild but specific inhibitor of only stage I of tumor promotion counteracted the action of TPA on gap junctions to some extent, which remained present in smaller numbers than in normal tissue at 24 h after the treatment. These results suggest that gap junctions are essential and specifically relevant to stage I tumor promotion.

Phorbol ester tumor promoter affects the mouse epidermal gap junctions.

Gap junctions are known to mediate cell to cell coupling. This study shows that the potent tumor promoter, 12-O-tetradecanoyl-phorbol-13-acetate (TPA), drastically affects the presence of gap junctions between mouse interfollicular epidermal (IFE) cells. In a time course ultrastructural study after a single application of 1 microgram of TPA the gap junctions between the IFE basal cells begin to decrease by 10 h post-exposure, are totally absent between 18 h and 30 h, and start reappearing at 30 h after TPA application. The potent hyperplasiogen , but very weak tumor promoter, mezerein, produces comparable hyperplasia and wide intercellular spaces but the population of gap junctions remains unchanged.

Effects of 12-O-tetradecanoylphorbol-13-acetate on the incorporation of labelled precursors into RNA, DNA and protein in epidermis, dermis and subcutis from precancerous mouse skin with reference to enhanced tumorigenesis.

The effects of a single application of 1.8 nmol 12-O-tetradecanoylphorbol-13-acetate (TPA) on precursor incorporation into RNA, DNA and protein in the epidermis, dermis and subcutis from 3-methylcholanthrene (MCA) injected precancerous mouse skin were studied at various time points between 3 and 96 h. In the precancerous tissues, the rates of incorporation of [3H]uridine into RNA did not alter appreciably from those in the control tissues; while the rates of [3H]methylthymidine incorporation into DNA were elevated with peaks appearing between 6 and 12 h, at 24 h and at 72 h in epidermis, dermis and subcutis. The rate of incorporation of [14C]leucine into protein was markedly elevated in all the three tissues which showed 3-4 sharp peaks. The maximum stimulation ranged between 14 and 20 times that of the control. A single application of TPA to the precancerous mouse skin induced early stimulation of precursor incorporation into all the three macromolecules in epidermis, dermis and subcutis. The increased stimulation was maintained for 36-72 h. The patterns of incorporation of [3H]methylthymidine into DNA gave rise to 2-3 peaks of elevated uptake in each tissue up to 36-48 h. A lowered rate of DNA synthesis between 48 and 60 h was followed by a peak at 72 h. In each group, epidermal mitotic activity correlated well with spurts of precursor incorporation into cellular DNA. The observations indicate that TPA recruits more cells into the DNA synthetic phase and accelerates selective growth of preneoplastic cells during tumor progression.

Dose response effect of retinyl acetate on DMBA induced carcinogenesis in the hamster cheek pouch.

The role of vitamin A in the induction and growth of chemically induced tumors is still controversial. While in some studies vitamin A influenced inhibition of chemically induced tumors was observed, other studies report on an enhancing influence of this substance on tumor induction. The cheek pouch epithelium of sixty five Syrian golden hamsters was exposed to DMBA and different doses of retinyl acetate, singly and in combination with DMBA. The findings suggest that 1) a significant delay occurs in tumor induction in animals treated with DMBA in combination with a higher concentration (2.0%) of retinyl acetate, 2) retinyl acetate at lower concentration (0.5 and 1.0%) does not inhibit or delay tumor induction.

Influence of excess of retinoid on DMBA carcinogenesis.

This paper reports a study on the influence of excess of vitamin A palmitate on the induction and maintenance of oral tumors. Sixty four weanling Syrian hamsters were divided in four groups and painted three times a week, either with 0.5% DMBA or 15% vitamin A palmitate, singly or in combination. A possible mild immune response evoked by vitamin A palmitate is considered responsible for the delayed induction of the tumors. In all animals exposed to carcinogen + vitamin A the induced tumors were small in size and histologically verified as well differentiated epidermoid carcinomas. In control group with sole vitamin A palmitate application the cheek pouch showed considerably increased keratinization.

12-0-Tetradecanoylphorbol-13-acetate-induced amplification of mesenchymal tumorigenesis in the mouse skin.

Skin tumors were induced in 6-week-old female Swiss albino mice by a single subcutaneous (SC) injection of 20-methylcholanthrene (MCA) in the right scapular region and the animals were then divided into four groups. Mice in group I did not receive further treatment. Six weeks after MCA injection, those in groups II and III received twice weekly applications of 0.1 ml acetone and 1.8 nmol 12-0-tetradecanoylphorbol-13-acetate (TPA) in 0.1 ml acetone, respectively, at the site of MCA injection until tumor development. Group IV animals were divided into four subsets and administered two, four, six, or eight TPA applications commencing 6 weeks after carcinogen injection. The effect of TPA pretreatment on MCA-induced tumorigenesis was studied in animals in group V. In mice treated with MCA alone, the most predominant mesenchymal tumor type is fibrosarcoma with induction of some rhabdomyosarcomas. Mixed mesenchymal tumors consisting of fibrosarcoma, rhabdomyosarcoma, or hibernoma were observed in only 12% of the animals. The number of animals bearing mixed mesenchymal tumors such as fibrosarcoma, rhabdomyosarcoma, hibernoma, and/or liposarcoma increased to 46% in mice receiving MCA + TPA until tumor development. Interestingly, liposarcomas were not found at all in animals treated with MCA alone. The data indicates that TPA application to precancerous mouse skin enhances mesenchymal tumorigenesis.