DMBA Treatment Research Articles

DNA damage induced by 7,12-dimethylbenz[a]anthracene in the liver and the mammary gland of rats exposed to polycyclic aromatic hydrocarbon enzyme inducers during perinatal life

The long-lasting modulating effect induced by the prenatal or neonatal exposure to phenobarbital (PB) and aroclor on the genotoxic activity of 7,12-dimethylbenz[a]anthracene (DMBA) in female Sprague-Dawley rats was studied. The effect was measured as DNA damage evaluated in the liver and in the mammary gland of 55-day-old animals, 4 and 24 h after an i.g. injection of 80 mg/kg of DMBA. PB was given per os, i.g. or in drinking water to pregnant females and by i.g. only to neonates or in adult progeny. Aroclor was injected i.g. in prenatal and in neonatal life, and a second dose was given in adult life. Under these experimental conditions it was shown that DNA damage kinetics caused by DMBA are modulated by exposure to PB and, to a minor extent, by aroclor. The amount and persistence of DNA damage were highest when PB was administered to neonates. An average 2-fold increase in the elution constants (K) of DNA in the liver and the mammary gland was observed 4 h after DMBA treatment, as compared to uninduced animals. Repeated enzyme induction by PB seems to reduce DMBA genotoxicity, as shown by a decrease in DNA damage and persistence in the liver and mammary gland. The inducibility of the monooxygenase enzyme system in perinatal life favouring metabolic activation or inactivation of polycyclic aromatic hydrocarbons might be critical in determining individual susceptibility of adult progeny to chemical carcinogenesis by DMBA.

Micronuclei in mouse skin cells following in vivo exposure to benzo[a]pyrene, 7,12‐dimethylbenz[a]anthracene, chrysene, pyrene and urethane

Detection of micronuclei (MN) in skin cells from HRA/Skh hairless mice treated with chemical or physical agents may prove informative in qualitative and quantitative studies of skin carcinogenesis. MN induction and cell survival were estimated in cytokinesis-blocked keratinocytes, cultured for 4 days in vitro, after a single topical dose of various organic compounds. Treatment with 2.56 micrograms (10 nmol) 7,12-dimethylbenz[a] anthracene (DMBA) resulted in maximal MN induction in cells removed from skin 12-24 hr after topical administration (79-88 MN/1,000 cells compared with 10-16 MN/1,000 cells in acetone-treated controls). Even in cells removed only 1 hr after DMBA treatment, a significant increase in MN was evident. However, to allow sufficient time for metabolic activation, a sampling time for of 24 hr was adopted for all test substances. Dose-dependent increases in MN were observed with DMBA, benzo[a]pyrene, chrysene, and urethane. Increased numbers of micronucleated cells were detected at the lowest doses administered in the present study (0.128, 0.5, 50, and 50 micrograms, respectively). Although reduced cell recovery occurred following exposure of mice to acetone, pyrene, and other chemicals, there was no evidence that cytotoxicity contributed to MN scored in keratinocytes. Moreover, the probable noncarcinogen, pyrene, failed to induce MN at doses from 2.5 micrograms to 2.5 mg/mouse. These results show that it is possible to assess chemical exposure in skin by measuring cell survival and skin genotoxicity by measuring MN induction in cultured keratinocytes. The available data suggest that MN induction may be a useful indicator of the carcinogenic potential of chemicals applied to the skin.

Lack of ecotropic virus involvement in induction of lymphomas in DBA/2J mice by 7,12-dimethylbenz(a)anthracene

DBA/2 mice carry a single endogenous ecotropic murine leukemia provirus, Emv-3, that is replication defective because of a single nucleotide substitution in codon 3 of p15gag. However, when weanling DBA/2 mice are treated percutaneously with 7,12-dimethylbenz(a)anthracene (DMBA), ecotropic virus replication is induced in almost all of the treated mice. Previous studies have shown that this induction results from DMBA-induced reverse mutations in codon 3 that allow efficient virus replication. In addition to ecotropic virus replication, DMBA also induces lymphomas in 100% of the treated mice. These results have raised the possibility that ecotropic virus replication is causally associated with the development of lymphomas in DBA/2 mice, perhaps via the insertional activation or mutation of cellular proto-oncogenes. To test this possibility, we compared lymphoma incidence after percutaneous DMBA treatment in DBA/2J-dv/dv mice, which carry two copies of Emv-3, with lymphoma incidence in DBA/2J-d+18J/d+18J mice, which lost both copies of Emv-3 by homologous recombination involving the long terminal repeat sequences. The results of this study conclusively demonstrated that Emv-3 is not causally associated with the development of DMBA-induced lymphomas in DBA/2J mice. Interestingly, histopathological and molecular analyses of the lymphomas indicated that the majority of the lymphomas in both strains of mice were of the B-cell lineage. This was unanticipated, since the majority of chemically induced lymphomas in other inbred strains are thymic lymphomas, presumably of the T-cell lineage. Thus, DBA/2 mice appear to present a unique model system for the investigation of chemically induced B-cell lymphomas in mice.

Protective effect of chorionic gonadotropin on DMBA-induced mammary carcinogenesis

The effect of the placental hormone chorionic gonadotropin (hCG) on 7,12-dimethylbenz(a)anthracene (DMBA)-induced mammary tumours was studied in young virgin Sprague-Dawley rats. This hormone when administered at a dose of 100 IU day-1 does not induce toxic effects, measured as alterations in body weight or weight of endocrine organs, and has a reversible effect on oestrous cycle. The lack of toxicity and the fact that hCG treatment terminated prior to administration of the chemical carcinogen DMBA protects the mammary gland from malignant transformation, led us to test the effect of hCG treatment on DMBA-initiated mammary tumours. Fifty day-old virgin Sprague-Dawley rats received intragastrically 8 mg DMBA per 100 g body weight and were divided into two groups: group I animals were treated with DMBA only and group II received DMBA at age 50 and in addition, a daily intraperitoneal injection of 100 IU hCG for days 21-81 post carcinogen administration. Tumorigenic response was evaluated by biweekly palpation of all animals and by complete autopsy 24 weeks after DMBA treatment. Group I animals developed an incidence of 100% of both tumours and adenocarcinomas. Group II animals developed a significantly lower incidence of tumours and adenocarcinomas, 51.5% and 45.5% respectively. In both groups lesions developed more frequently in thoracic than in abdominal mammary glands. It is postulated that hCG treatment, probably through stimulation of ovarian oestrogen and progesterone synthesis, induces differentiation of mammary epithelium that although affected by the carcinogen can still be rescued from malignant transformation.

Oncogene alterations in in vitro transformed rat tracheal epithelial cells

10 derivations of rat tracheal epithelial (RTE) cells, including normal cells, normal primary cultures, 7 tumorigenic cell lines and 1 nontumorigenic cell line transformed in vitro by treatment with 7,12-dimethyl-benz[ a]anthracene (DMBA), benzo[ a]pyrene (BP) and/or 12- O-tetradecanoylphorbol-13-acetate (TPA) were examined for oncogene alterations. No abnormalities of Ha- ras were seen that were suggestive of amplification, rearrangement or the presence of RFLPs. Analysis of specific-point mutations in Ha- ras using Pst I digestion (codon 12, GGA to GCA) or Ha- ras and Ki- ras using Xba I (codon 61, CAA to CTA) were negative. In one cell line derived by DMBA treatment, changes in the c- myc restriction digest pattern were seen after incubation with Bam HI and Hind III. Northern analysis revealed consistent differences between normal and transformed cells when probed with Ha- ras; c- myc expression was of low intensity, and the expression of Ki- ras could not be detected. Transfection of RTE cell DNAs into NIH/3T3 cells did not result in the appearance of morphologic transformants. The studies suggest that Ha- ras or Ki- ras codon 61 A to T transversions (CAA to CTA) are not associated with the immoral/tumorigenic phenotype in RTE cells transformed by DMBA or TPA, and are in contrast to results reported in some other biological systems.

Tissue‐specific activating mutations of Ha‐ and Ki‐ras oncogenes in skin, lung, and liver tumors induced in mice following transplacental exposure to DMBA

Transplacental carcinogenesis represents a good model in which to study the involvement of tissue-specific oncogene activation in carcinogenesis because a single exposure to a carcinogen induces tumors at various sites. We tested tumors of the skin, liver, and lung produced in mice after transplacental 7,12-dimethylbenz[a]-anthracene (DMBA) exposure for possible activation of ras genes. XbaI restriction fragment-length polymorphism analysis has shown that exposure to DMBA in utero may result in appearance of A----T transversion at the second position of codon 61 of Ha-ras oncogene in skin and liver tumors but not in lung tumors. Moreover, DNA samples isolated from spontaneous and DMBA-induced lung and liver tumors were analyzed for mutations at the same position of Ki-ras oncogene using differential hybridization with specific oligonucleotides. Among five spontaneous lung tumors, three cases of A----G transition, and one case of A----T transversion were found, whereas four of ten lung tumors of DMBA-treated animals were positive for A----T mutation. No Ki-ras mutation was detected in one spontaneous and four DMBA-induced hepatomas. In two cases, we revealed Ki-ras A----T mutation in the lung tumor and Ha-ras mutation in the liver tumor taken from the same animal. These results indicate first that DMBA treatment may induce A----T mutation at the second position of codon 61 both in Ha-ras and in Ki-ras and, second, that the role of different activated oncogenes in carcinogenesis may differ, depending on the tissue in which the tumor develops.

Immunophenotypic and cell cycle analysis of lymph node cells from dimethylbenzanthracene-treated mice

The chemical carcinogen 7, 12-dimethylbenz-(a)anthracene (DMBA) depletes Langerhans cells from murine epidermis. Application of contact sensitizers to DMBA-treated skin induces specific immunological tolerance due to a DMBA-resistant epidermal antigen presenting cell (APC) migrating to local lymph nodes where it presents antigen in a way which activates suppressor cells. As alterations in local lymph node lymphocytes may enhance the ability of the DMBA-resistant APC to activate suppressor cells, these cells were examined in DMBA-treated mice. Lymph nodes in DMBA-treated mice had normal morphology but were larger and contained increased numbers of lymphocytes. Cell cycle analysis revealed that these lymphocytes did not arise from division within the lymph node, suggesting alterations in homing properties of lymphocytes. Contact sensitizer applied to DMBA-treated skin did not increase lymphocyte division, possibly due to suppressor cell inhibition of the development of effector lymphocytes. DMBA treatment had no effect on B cells or Ia expression, but decreased levels of the T lymphocyte cell surface molecule Thy-1, and increased L3T4 and Lyt-2 as quantitated by flow cytofluorimetry. These changes could influence the development of immune responses as these T cell molecules are receptors involved in lymphocyte interactions.

Herpes simplex virus enhances the 7,12-dimethylbenz [a]anthracene (DMBA)-induced carcinogenesis and amplification and overexpression of c-erb-B-1 proto-oncogene in hamster buccal pouch epithelium

We studied the effects of herpes simplex virus type 1 (HSV-1) inoculation and topical 7, 12-dimethylbenz [a]anthracene (DMBA) application, alone or in combination, on the carcinogenesis and on the amplification and expression of various cellular proto-oncogenes in hamster buccal pouch tissue. Topical DMBA treatment produced tumor formation in pouches, but HSV-1 inoculation alone caused no neoplastic changes. In pouch tissues receiving both DMBA application and HSV-1 inoculation, the development of initial leukoplakia and tumor has hastened and enhanced in comparison with those receiving DMBA alone. Topical DMBA application to pouch tissue induced an amplification and an increase in the expression of cellular erb-B-1 (c-erb-B-1) proto-oncogene in the epithelial tissue, whereas repeated infection with HSV-1 alone did not. Topical DMBA combined with HSV-1 inoculation, however, resulted in greater amplification and expression of c-erb-B-1 proto-oncogene in the pouch epithelial tissue compared to the DMBA alone. These data indicate that HSV-1 inoculation significantly increases the carcinogenic activity of DMBA, in part, by probably enhancing DMBA-induced amplification and expression of c-erb-B-1 proto-oncogene in hamster buccal pouch tissue.

Effect of DMBA on oral cancer development in hamsters with latent HSV-1 infections in trigeminal ganglia

This study examined the effect of 7, 12-dimethyl-benz(a)anthracene (DMBA), polycyclic aromatic hydrocarbon carcinogen, on pathologic changes in buccal pouch mucosa of hamsters with latent herpes simplex virus type 1 (HSV-1) infections in their trigeminal ganglia. Of the pouches receiving DMBA treatment, the average number of tumors per pouch was significantly higher ( p < 0.01) in pouches of animals with HSV-1 infections as compared to those that received DMBA only. Of the pouches receiving DMBA, the average cumulative tumor diameter (the sum of the tumor diameters in each group divided by the number of pouches in that group) was notably greater in animals with latent HSV-1 than in animals that had not been infected; however, this difference was not significant. The DMBA-treated hamsters that had latent infections also displayed a higher severity and prevalence of histopathologic changes in their pouch mucosa. This study indicates that latent HSV-1 and DMBA show synergism in the development of tumors and precancerous histopathologic changes in hamster buccal pouch mucosa. An unexpected finding was that among the HSV-1 infected animals, 95% of the ganglia from animals treated with DMBA showed latent HSV-1 virus on explanation culture, whereas only 10% of the ganglia from infected animals that received mineral oil, rather than DMBA, contained latent virus.

Acceleration of papilloma growth in mice fed high‐fat diets during promotion of two‐stage skin carcinogenesis

The effect of feeding a high-fat diet during the promotion phase of skin tumorigenesis was assessed in SENCAR mice. Tumors were initiated on the backs of mice by application of 10 nmol 7,12-dimethylbenz[a]anthracene (DMBA); the tumors were then promoted beginning one week later with twice weekly treatments of 2 micrograms 12-O-tetradecanoylphorbol-13-acetate (TPA) each in 0.2 ml acetone. Control diet containing 5% corn oil was fed from four weeks before until one week after DMBA treatment in all groups. A high-fat diet (24.6% corn oil) was fed from one week after DMBA treatment; the other groups continued on the control diet. Mice were fed ad libitum, and those given the high-fat diet consumed more calories early in the study than the controls did. Treatment with TPA increased calorie consumption throughout the study. Body weights were elevated in mice fed high-fat diets and reduced by TPA treatment. The average number of papillomas per mouse did not differ between the low- and high-fat groups, but papillomas grew more rapidly on the mice fed a high-fat diet than those fed a low-fat diet. The feeding of a high-fat diet following DMBA treatment, in the absence of TPA administration, did not result in promotion of skin tumors. Therefore, a high-fat diet acted as a copromoter of skin tumors in SENCAR mice.

Chemoprevention of experimental mammary carcinogenesis by the synthetic organoselenium compound, benzylselenocyanate, in rats

The effect of the dietary organoselenium compound, benzylselenocyanate (BSC) along with its sulphur analogue, benzylthiocynanate (BTC) and sodium selenite (Na2SeO3), on 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary carcinogenesis was examined in female Sprague-Dawley rats during the initiation phase of carcinogenesis. Semipurified diets containing 25 p.p.m. of BSC and 25 p.p.m. BTC, and 4 p.p.m. Selenium as Na2SeO3 in drinking water were given to 5-week-old rats for 3 weeks starting 2 weeks before, during and until 1 week after carcinogen treatment. At 7 weeks of age animals were given a single dose of DMBA (10 mg) in 1 ml olive oil by oral intubation. One week after DMBA treatment, the groups receiving BSC- and BTC-supplemented diets were transferred to the unsupplemented standard diets and the group of rats receiving Na2SeO3 in drinking water was transferred to regular tap water for the duration of the experiment. The results indicate that the rats receiving BSC in their diet showed a highly significant inhibition of tumor incidence and tumor multiplicity as well as a prolonged latency period when compared to the group fed the control diet. Neither BTC nor Na2SeO3 had any effect on the subsequent development of mammary tumors. These results indicate that dietary BSC inhibits mammary tumor incidence during the initiation phase of carcinogenesis and is a considerably more potent inhibitor than its sulphur analogue BTC and inorganic selenium. This is the first report that demonstrates the inhibition of mammary carcinogenesis by a synthetic organoselenium compound.

Prevention of mammary carcinogenesis in rats by pregnancy: effect of full-term and interrupted pregnancy.

In this study, the role of parity in conferring protection of the mammary gland against chemical carcinogenesis induced by 7,12-dimethylbenz(a)anthracene (DMBA) was investigated. Experiments were also carried out to determine if an 'interrupted' pregnancy was capable of reducing the incidence of mammary tumour induction. Since it has been suggested that morphological development or the proliferative pattern of the mammary gland at the time of carcinogen administration may be involved in reducing the susceptibility of the mammary gland to chemical carcinogenesis, experiments were designed to elucidate the possible influence of these two factors. Sprague-Dawley female rats were mated and were either allowed to complete pregnancy and parturition or were subjected to Caesarian section on day 5, 10 or 15 of the pregnancy. When DMBA was administered i.v. to animals which had been allowed to complete a full-term pregnancy, only 14% developed tumours, compared to 70% in age-matched nulliparous controls. Termination of the pregnancy on days 5, 10 or 15 was as effective in reducing tumour incidence as full-term gestation and parturition, but still resulted in partial and statistically significant inhibition, compared to age-matched nulliparous controls. There was no significant difference in 3H-thymidine labelling index (LI) at the time of DMBA treatment in the parous rats compared to age-matched nulliparous controls. We also observed no significant differences in the morphological development of the mammary gland in parous and nulliparous rats of the same age. These results indicate that the protective mechanism may not lie in the mammary gland per se, but may indeed be a host factor, such as hormonal or immunological changes occurring in the host as a result of the pregnancy.

Arteriosclerotic plaque development is ‘promoted’ by polynuclear aromatic hydrocarbons

In previous work we found that weekly injections of the polynuclear aromatic hydrocarbon (PAH) carcinogen 7,12-dimethylbenz[a]anthracene (DMBA) induced spontaneous aortic plaques in cockerels to grow to a larger size and at a faster rate than plaques in control animals. To determine whether plaque-stimulating ability is related to carcinogenic potency or mutagenicity we have now tested a variety of agents, including PAH carcinogens, non-PAH carcinogens and weakly carcinogenic PAHs. Cockerels were injected weekly (from 4-20 weeks of age) with one of the following compounds: benzo[a]pyrene (B[a]P), benzo[e]pyrene (B[e]P), dibenz[a,h]anthracene (AH), dibenz[a,c]anthracene (AC), 3-methylcholanthrene (MCA), acetylaminofluorene (AAF), N-methyl-N,N'-nitro-nitrosoguanidine (MNNG) or anthracene (ANT). Plaques were present in the abdominal aortas of all animals. Plaque volumes were 8-14 times greater in AC-, B[a]P-, B[e]P-, MCA- and AH-treated cockerels than in controls. Plaques were slightly larger in the AAF-treated group than in control animals, and in the ANT- and MNNG-treated groups were indistinguishable in size from plaques in control animals. The largest plaque volumes were in AH-treated cockerels and were comparable in size to those elicited by DMBA treatment. The accelerated development of plaques is consistent with a 'promotional' role for these agents. There was a poor correlation between mutagenicity or carcinogenicity and plaque 'promotion', which may reflect a role for different metabolites in these processes.

The influence of a hyperthermia treatment on chemically induced tumor initiation and progression in mouse skin.

A single hyperthermia treatment given near the time of initiation with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or 7,12-dimethylbenz[a]anthracene (DMBA) increased the number of initiated cells in the skin of SENCAR mice subjected to a two-stage tumorigenesis protocol. In animals subsequently promoted with 12-O-tetradecanoylphorbol-13-acetate (TPA), a 44 degrees C, 30-min hyperthermia treatment given just before, just after or 24 h before MNNG initiation increased the average papilloma frequency by 40-50%. In the groups of animals that received a hyperthermia treatment just before MNNG initiation, tumor latency was reduced by 40-60%. Treatment with MNNG in the absence of hyperthermia produced two classes of initiated cells. One type, formed in low yield, was independent of TPA promotion and formed tumors with a high probability of progression to malignancy. The other type was promotion dependent, and formed in relatively high yield but produced tumors with a probability of progression to carcinomas approximately 10-fold less than promotion-independent initiated cells. A single hyperthermia treatment given just before or just after MNNG initiation increased the yield of both promotion-dependent and promotion-independent initiated cells, and consequently increased the yield of carcinomas. In animals given a single hyperthermia treatment 24 h prior to initiation (to induce thermotolerant skin cells), MNNG exposure resulted in an increased yield of promotion-dependent initiated cells but no change in the yield of promotion-independent initiated cells. Hyperthermia treatment of DMBA-initiated skin increased the yield of initiated cells (promotion-dependent) only when given just after exposure to the initiator. The extra initiated cells produced by hyperthermia treatment of MNNG or DMBA exposed skin had the same probability of progression to carcinomas as initiated cells produced by the same initiation in the absence of hyperthermia. As noted previously for DMBA-initiated mice, hyperthermia given at the time of each application of TPA promoter also suppressed the formation of papillomas initiated by MNNG. Only the promotion and progression of promotion-dependent initiated cells, and not of promotion-independent cells, was suppressed. The results show that a single hyperthermia treatment near the time of exposure to an alkylating agent increased the number of both promotion-dependent and promotion-independent initiated cells and, as a consequence, increased the risk of carcinogenesis associated with that exposure.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of 7,12-dimethylbenz[a]anthracene, benzo[a]pyrene and cyclosporin A on murine cytomegalovirus infection: Studies of resistance mechanisms

Susceptibility to murine cytomegalovirus (MCMV) was enhanced by treating B 6C 3F 1 and CD-1 mice subcutaneously with 100 mg 7,12-dimethyl-benz[a]anthracene (DMBA)/kg fractionated over a 2 week period prior to sub-lethal infection. Virus-augmented natural killer cell (NKC) activity was depressed in B 6C 3F 1 mice treated with 100 mg DMBA/kg, while serum interferon (IFN) levels were unaffected. Treatment with 50 mg DMBA/kg had no effect on susceptibility to virus or virus-augmented NKC activity. Susceptibility to MCMV was not affected by treating mice with 400 mg benzo[a]pyrene (B[a]P)/kg using the same exposure regimen. Virus-augmented NKC activity was suppressed in B[a]P)-treated mice, but the magnitude of the suppression (18%) was much less than that for DMBA-treated mice (39%). Susceptibility to MCMV, virus-augmented NKC and IFN induction were not affected in mice treated intraperitoneally with 50 mg cyclosporin A (CSA)/kg/day for 5 days and infected on the 5th day of treatment. In contrast, enhanced susceptibility to MCMV and depressed NKC activity were observed in mice treated by the same exposure regimen on days 1–5 post infection. Susceptibility was not affected by CSA given on days 5–9 post infection. The data are useful not only because they show that DMBA and appropriately-timed CSA treatments suppress virus augmented NKC and enhance susceptibility to MCMV, but also because they help to define the relative importance of certain immune responses in defending against the infection, thus improving the usefulness of MCMV as a host resistance model for immunotoxicity testing. The data suggest that chemicals which depress NKC are likely to enhance susceptibility to MCMV, and conversely that effects on NKC should be suspected when chemical exposure enhances susceptibility to MCMV.

Early neoplastic commitment of hamster buccal pouch epithelium exposed biweekly to 7,12-dimethylbenz[a]anthracene.

Experiments were performed to determine the rate at which hamster buccal pouch epithelium (HBPE) is committed to neoplastic development during a regimen of biweekly topical applications of 7,12-dimethylbenz[a]anthracene (DMBA) in mineral oil, administered for 1, 2, 3, 5 or 10 weeks. Evaluated indices of neoplastic commitment were: (i) primary tumor formation, (ii) passageability of HBPE cells in surface culture, (iii) anchorage-independent growth in soft agar and (iv) induction of the transformed phenotype in NIH3T3 cells following transfection with HBPE DNA. Groups of 12-15 hamsters, exposed to DMBA for 1, 2 and 3 weeks, developed buccal pouch tumor incidences of 13%, 42% and 71% respectively within 44 weeks. Buccal pouches of ten control hamsters treated for three weeks with mineral oil did not develop buccal pouch neoplasms during an observation period of 44 weeks. Whereas cultured HBPE cells obtained following three weeks of in vivo DMBA exposure were negative in assays for anchorage-independent growth, passageability in surface culture and induction of NIH3T3 transformants following DNA transfection, similarly cultured cells obtained following five and ten weeks of in vivo exposure were positive, or marginally positive, in each of these assays. These results indicate that (i) the regimen of biweekly DMBA applications commits HBPE to neoplastic development within three weeks and that (ii) subsequent cellular or molecular changes occurring during greater than or equal to 2 succeeding weeks of DMBA treatment are necessary to manifest the transformation associated phenotypes of continuous passageability, anchorage-independent growth, and induction of NIH3T3 transformants following DNA transfection.

The effect of indomethacin on tumor regression in DMBA-induced epithelial neoplasia of hamster cheek pouch mucosa

Tumors were allowed to develop in the cheek pouches in two groups of hamsters following topical application three times per week of a 0.5% solution of 7, 12-dimethylbenz(α)anthracene (DMBA) in liquid paraffin administered for 10 weeks. Two control groups received no DMBA treatment. After carcinogen application, the animals in one DMBA-treated group and those in one of the control groups were given indomethacin daily; this was administered through the oral route for an additional 10 weeks. At this time, all of the animals were killed and their cheek pouches, the draining cervical lymph nodes, and other organs were removed for examination. The number and size of tumors in each pouch were recorded, and each pouch was weighed. Indomethacin, in the dosage administered, was not found to cause regression of established tumors. The effects of prostaglandins, indomethacin, and other prostaglandin-synthesis inhibitors in tumor tissue are discussed. It is suggested that more knowledge is needed about the role of prostaglandins, and the related products of AA metabolism, in squamous cell cancer before indomethacin is used therapeutically.

Low Level X-Radiation Effects on Carcinogenesis by 7,12-Dimethylbenz(a)anthracene in Syrian Hamster Cheek Pouch Epithelium: Acute vs Fractionated Radiation Dose Studies

Studies examined the effects of acute and fractionated low to moderate level X-ray exposures on hamster cheek pouch carcinogenesis in vivo by 7,12-dimethylbenz(a)anthracene (DMBA). Animals were grouped by treatment as follows: acute doses of 0.85-3.40 Gy X rays; 17 once weekly doses of 0.01-0.20 Gy X rays (fractionated radiation); topical DMBA for 10 weeks; DMBA plus fractionated radiation starting together; DMBA plus acute radiation in Week 1 or 10 of DMBA treatments; and sham irradiation, DMBA vehicle, or anesthesia controls. After 44 weeks, hamsters were sacrificed, and their cheek pouches were excised, serially sectioned, and examined by light microscopy for histopathology. No histologic changes were observed in radiation-only hamsters. Carcinoma incidences in DMBA-only groups ranged from 45 to 60%. Carcinoma incidences were greater in groups receiving DMBA plus fractionated radiation than in groups receiving either acute radiation + DMBA or DMBA alone. Carcinoma incidences in acute radiation plus DMBA groups were lower than those in DMBA-only groups. These results suggest complex interactions between radiation and DMBA, perhaps with radiogenic cell killing being a principal factor in acute radiation + DMBA groups, and reciprocal additive or synergistic effects of radiation and DMBA on cancer induction and manifestation in fractionated radiation + DMBA groups.

ＤＭＢＡ処理により誘発されたラット舌の組織学的変化に伴う味覚応答に関する研究

ＤＭＢＡ処理により誘発されたラット舌の組織学的変化に伴う味覚応答に関する研究

Prolactin cells of female rats treated with the carcinogen 7,12-dimethylbenz(A) anthracene (DMBA) in vivo. An ultrastructural study.

Pituitary prolactin (PRL) cell microadenomata developed in females of a strain of Sprague-Dawley rats (SD1) following intragastric treatment with the carcinogenic hydrocarbon 7,12-dimethylbenz(a) anthracene (DMBA). Similar treatment of another strain of Sprague-Dawley rats (SD2) resulted in mammary tumour development but no PRL cell microadenomata. In SD1 strain rats morphologically distinct populations of PRL cells appeared after DMBA treatment, one composed of cells characterised by abundant, organised but very dilated RER and with large hormone storage granules, 500-600 nm in diameter (P1). The other cell type had electron-dense cytoplasm, narrow organised arrays of RER and moderately large, pleomorphic granules (P2). Both cell types appeared active with large Golgi and prominent nucleoli. P2 cells were most numerous 2 months after DMBA treatment but had virtually disappeared at 6 months and microadenomata were common at 8 months. PRL cells of SD2 rats were uniform in morphology, characterised by only moderate accumulations of RER, pleomorphic hormone storage granules, large Golgi and prominent nucleoli, and showed no close resemblance to either P1 or P2 cells of SD1 strain rats. It is possible that the morphological variations which developed in SD1 PRL cells may represent changes in responsiveness to factors controlling PRL cell secretion and proliferation and which may be pertinent to microadenoma development.