PubMed Health⌕ Search

Biomedical subjects

I B Gimenez-Conti

Publications and source records attributed to I B Gimenez-Conti.

30 records · Page 2Linked to original sources
PubMed1993

The hamster cheek pouch carcinogenesis model.

The Syrian golden hamster cheek pouch carcinogenesis model is probably the best-known animal system that closely compares to events involved in the development of premalignant and malignant human oral cancers. Furthermore, it is one of the most well-characterized models for squamous cell carcinomas (SCCs). However, stages of carcinogenesis (initiation, promotion, and progression) have not been well-defined in this system. Basic understanding of the mechanism(s) of carcinogenesis in this organ is instrumental for the development of new strategies for chemoprevention and early chemointervention. To understand the important early events that occur in the hamster cheek pouch carcinogenesis model, we compared it to the mouse skin model, where a number of critical events have been well characterized. We determined that approximately 60% of the hamster cheek pouch SCCs have a mutation in codon 61 of the Ha-ras gene. We also established a two-stage carcinogenesis protocol in this model using a single dose of dimethylbenz(alpha)anthracene (DMBA) and multiple doses of benzoyl peroxide for 45 weeks. Twenty-five percent of tumors developed with this protocol had the same mutation in codon 61 of the Ha-ras gene, confirming that this mutation, as in the mouse skin model, is initiation-related. We examined the sequential expression of hyperplasia, micronucleated cells, ornithine decarboxylase (ODC) activity, polyamine levels, transglutaminase I activity, epidermal growth factor receptor (EGF-R) levels, keratins, gamma-glutamyltranspeptidase (GGT), transforming growth factor-beta 1 (TGF-beta 1), leukoplakia, and carcinomas induced during carcinogenesis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Low frequency of codon 61 Ha-ras mutations and lack of keratin 13 expression in 7,12-dimethylbenz[a]-anthracene-induced hamster skin tumors.

Alterations in the pattern of keratin expression are a common feature of skin-tumor development. In this study, we investigated whether the loss of epidermal keratin 1 (K1) and its replacement by mucosal keratin 13 (K13) is unique to mouse skin tumors induced by 7,12-dimethylbenz[a]anthracene (DMBA) and 12-O-tetradecanoylphorbol-13-acetate (TPA), since it has been reported that human epidermal tumors do not exhibit aberrant expression of K13. With that purpose, we analyzed the keratin profiles of 16 DMBA-induced hamster skin tumors using monospecific antibodies against K1 and K13. Although all the tumors expressed K1, they also showed an overall tendency towards loss of this keratin; furthermore, none of the tumors expressed K13. Previous studies have suggested that the induction of K13 in mouse skin is related to the mutation of the Ha-ras gene by the initiating agent DMBA, a mutation consistently found in murine DMBA/TPA-induced tumors and rarely found in human skin tumors. Therefore, we also evaluated the tumors for the presence of codon-61 mutations by direct sequencing of DNA extracted from paraffin-embedded tissue sections. Only three tumors showed an A-->T transversion in the second nucleotide of Ha-ras codon 61. However, presence of the mutation did not correlate with K1 staining. Although hamster skin tumors were induced by the same initiator as were mouse skin tumors, hamster skin tumors did not show the same keratin profile. Moreover, their immunohistochemical expression of K1 and K13 and their codon 61 sequences resembled that of their human counterparts. These results suggest that the aberrant expression of K13 may be unique to murine skin. Furthermore, although codon 61 Ha-ras mutation appears to be related to keratin alterations in the mouse model, this mutation is not sufficient to produce the same biochemical changes in other species.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1993

Morphological changes of the nucleolar organizer regions induced by 7,12-dimethylbenz[a]anthracene in the hamster cheek pouch.

The staining of nucleolar organizer regions (NORs), which equate to rDNA transcription, was applied to chemically induced-lesions of the hamster cheek pouch. The cheek pouches of 16 male golden Syrian hamsters were treated three times a week with 0.5% 7,12-dimethylbenz[a]anthracene (DMBA) in mineral oil for 16 weeks. The percentage of gathered-type NORs with high activity nucleoli increased in the pouch epithelium during DMBA treatment and reached the highest values in malignant tumors. The percentage of dispersed-type NORs also increased in the malignant lesions. However, the absolute number of NORs was not affected by DMBA treatment. These results suggest that DMBA induces modification of NOR activity at the early stages of carcinogenesis and shows the potential of this model for studying NOR alterations in neoplasia.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1992

Dissociation of sensitivities to tumor promotion and progression in outbred and inbred SENCAR mice.

The sensitivity of outbred SENCAR mice and inbred SENCAR (SSIN) mice to multistage carcinogenesis was studied. Tumors were induced using either 7,12-dimethylbenz[a]anthracene or N-methyl-N'-nitro-N-nitrosoguanidine as initiators and 12-O-tetradecanoylphorbol-13-acetate or benzoyl peroxide as promoting agents. Although the number of papillomas per mouse was higher in SSIN than in outbred SENCAR mice, the number of carcinomas observed in the SSIN strain was significantly lower regardless of the initiator or promoter used. It was also observed that the expression of markers of premalignant progression (i.e., dysplasia, expression of keratin K13, and loss of keratin K1 expression) was markedly suppressed in SSIN papillomas. After 50 wk of promotion with 12-O-tetradecanoylphorbol-13-acetate, the pattern of expression of K13 and K1 in SSIN mice was comparable to the pattern observed in outbred SENCAR mice after 10 to 20 wk of promotion with 12-O-tetradecanoylphorbol-13-acetate. It was also observed that 67% of the tumors induced in SSIN mice by initiation with 7,12-dimethylbenz[a]anthracene exhibited a mutation in codon 61 of the Ha-ras-1 gene. This latter finding suggests that the differences observed in tumor progression between the inbred strain and the outbred stock are not related to a genetic alteration in the Ha-ras-1 gene but rather to an independent event that we have postulated to involve a putative suppressor gene. The data reported here suggest that the putative gene(s) that confers susceptibility to tumor promotion was segregated from the gene(s) involved in tumor progression during selection and inbreeding of the SENCAR mouse stock.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1992

Activating mutation of the Ha-ras gene in chemically induced tumors of the hamster cheek pouch.

The presence of an activating mutation in the Ha-ras gene in hamster cheek pouch tumors induced by 7,12-dimethylbenz[a]anthracene (DMBA) complete carcinogenesis was investigated. The normal sequence of a fragment of genomic DNA encompassing codon 61 of the Ha-ras gene was amplified by the polymerase chain reaction using primers designed for a highly conserved region of the mouse Ha-ras-1 gene. The sequence of the amplified fragment was determined by a direct sequencing technique and exhibited 83.3% and 87.5% homology with the corresponding human and mouse sequences, respectively. At the amino acid level, the sequence was identical among the three species. Paraffin sections of 11 squamous cell carcinomas of the cheek pouch were used to detect mutated Ha-ras alleles. DNA sequencing of the tumors showed that six of 11 tumors presented an A----T transversion in the second position of codon 61, resulting in an amino acid change from glycine to leucine. As has been demonstrated in other systems, we have shown a specific mutation of the Ha-ras gene in chemically induced tumors of the hamster cheek pouch, further supporting the role of this oncogene in chemical carcinogenesis.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1992

An animal model for oral cancer.

Human head and neck squamous cell carcinogenesis (SCC) is a common malignancy that appears to be related to continuous exposure to putative carcinogens or promoters such as tobacco and alcohol. To understand the mechanisms of the development of head and neck cancer and to test the efficiency of new therapeutic approaches, the characterization of an animal model system is necessary. The check-pouch carcinogenesis model in Syrian golden hamsters is probably the best known animal system that is most closely comparable with the development of premalignant and malignant lesions in human oral cancer. Furthermore, it is one of the most well-characterized animal system models for SCC. Our first approach to understanding the cellular and molecular changes that occur in the hamster cheek-pouch carcinogenesis process was to compare this model to the mouse-skin system, in which a number of critical events have been well characterized. We examined the sequential expression of hyperplasia, micronucleated cells, ornithine decarboxylase activity, polyamine levels, transglutaminase I activity, epidermal growth factor receptor levels, expression of several keratins, gamma-glutamyl transpeptidase, and nucleolar organizer regions. We suggest that these markers can be used to understand mechanisms of carcinogenesis and, in addition, can serve as alternative shorter end points in studies of chemoprevention. We also present preliminary molecular studies in the experimental oral model. We obtained a partial sequence of exon 2 of the Ha-ras gene and detected an A182----T transversion in codon 61 in hamster cheek-pouch SCC induced by 7,12-dimethylbenz(a)anthracene.

Animals↗
PubMed1990

Changes in keratin expression during 7,12-dimethylbenz[a]anthracene-induced hamster cheek pouch carcinogenesis.

This study was undertaken to explore the expression of keratins in the hamster cheek pouch carcinogenesis model, using monospecific keratin antibodies and a technique that allows immunoblotting analysis of tissues embedded in paraffin. Changes in keratin expression were correlated with histopathological changes and with the expression of the enzyme gamma-glutamyl transpeptidase. The right cheek pouch of 20 male golden Syrian hamsters was treated with 0.5% 7,12-dimethylbenz[a]anthracene for 16 weeks. As previously described by other laboratories, this treatment resulted in hyperplastic and dysplastic lesions and benign and malignant tumors. The keratins assayed in this study were K14 (Mr 55,000), K1 (Mr 67,000), and K13 (Mr 47,000). The normal hamster cheek pouch epithelium expressed K14 in the basal layer and K13 in the suprabasal and differentiated layers, whereas K1 was not detected by either immunohistochemistry or immunoblotting. Concomitant with 7,12-dimethylbenz[a]anthracene-induced hyperplasia, there were some topographical alterations in the distribution of K14. In this case, K14 was no longer restricted to the basal layer but was also expressed in differentiated cells. The same pattern was also observed in dysplastic lesions and in squamous cell carcinoma. Furthermore, expression of the K13 differentiation-associated keratin was preserved in this hyperplastic epithelium during all the stages of carcinogenesis, including either anaplastic or differentiated areas. In contrast, after 2 weeks of 7,12-dimethylbenz[a]anthracene treatment, K1 expression started as a weak and patchy pattern in suprabasal cells, becoming stronger and more homogeneous at 8 and 16 weeks of treatment. However, K1 was almost absent in squamous cell carcinoma, where only small very well differentiated areas were stained. We also observed gamma-glutamyl transpeptidase-positive foci in earlier stages of carcinogenesis, concomitant with the expression of the K1 keratin. However, it was not possible to find a perfect topographical correspondence between the two events. Alterations in the pattern of keratin expression appear to be a common feature during the development of squamous cell carcinoma in different systems and could be an excellent tool to study carcinogenesis and chemoprevention.

9,10-Dimethyl-1,2-benzanthracene↗
PubMed1984

Estrogen and progesterone regulation of proliferation, migration, and loss in different target cells of rabbit uterine epithelium.

We have explored the possibility that estrogens and progesterone could have different target uterine cell populations according to their cell cycle stage and localization in glands vs. lumen. Experiments were carried out in which rabbits were injected with [3H]thymidine for 3 days to label nuclei of dividing cells, then either 17 beta-estradiol, progesterone, or vehicle were administered. 17 beta-Estradiol induced a decrease in the percentage of cells with labeled nuclei or labeling index of either luminal or glandular epithelium. Since this steroid has been shown to have a significant proliferative effect on glands, the data suggest that its effect is exerted on unlabeled quiescent cells, which are then recruited into the cell cycle. Progesterone, on the other hand, was found to induce a significant increase in labeling index of both luminal and glandular epithelium. Therefore, it is concluded that dividing cells are a target for this hormone. Analysis of the number of nuclear grains according to cell location in luminal vs. glandular epithelia and the effect of hormone administration confirmed that each ovarian hormone acts on different target cell populations. Short and long term administration of estrogens resulted in a larger internal circumference of the uterus due to an increase in the number of luminal cells, whereas the number of glands and glandular cells per section did not appear to change. These findings, in combination with previous research, suggest that endometrial gland cells migrate towards the lumen and estrogen administration decreases the rate of cell loss in the luminal epithelium. The concept of cell migration is supported by experiments in which single administration of [3H]thymidine to rabbits was followed by determination at different times of the geographical distribution of cells with labeled nuclei. There was observed, as a function of time, a decrease in the number of labeled cells in the bottom of the glands with a concomitant increase in the same parameter in the upper part of the glands and luminal epithelia. Estradiol administration changed these kinetics.

Animals↗