Biomedical subjects

R E Mitchel

Publications and source records attributed to R E Mitchel.

At least 37 records · Page 2Linked to original sources

PubMed1994

Radiation-induced adaptive response for protection against micronucleus formation and neoplastic transformation in C3H 10T1/2 mouse embryo cells.

We have monitored the end points of cellular survival, micronucleus formation and neoplastic transformation frequency to assess adaptation to ionizing radiation in the C3H 10T1/2 mouse embryo cell system. Plateau-phase cells were pre-exposed to an adapting dose of 0.1 to 1.5 Gy low-dose-rate gamma radiation 3.5 h prior to an acute challenge dose of 4 Gy. No adapting dose improved clonogenic survival detectably, whether the cells were plated immediately after the acute exposure or held in plateau phase for 3.5 h before plating. However, all chronic adapting doses resulted in both a reduction in micronucleus frequency in binucleate cells and about a twofold reduction in neoplastic transformation frequency per viable cell when cells were subsequently exposed to the 4-Gy challenge dose. Our data suggest that a low-dose-rate pre-exposure to ionizing radiation induces an adaptive response in C3H 10T1/2 cells, and that this response enhances DNA double-strand break repair when cells are subsequently exposed to a second radiation dose. This enhanced repair appears to be error-free since these adapted cells are also less susceptible to radiation-induced neoplastic transformation.

Adaptation, Physiological↗

PubMed1993

Effects of taxol and taxol/hyperthermia treatments on the functional polarization of cytotoxic T lymphocytes.

Immunofluorescence staining, electron microscopy, and (51Cr) cytolytic release assays are used to investigate the effects of taxol and taxol/hyperthermia treatments on the microtubule organization and cytolytic activity of cytotoxic T lymphocytes (CTLs). A 4 h treatment of CTLs with 1 microM taxol results in an extensive reorganization of the microtubule system to form one to a few large microtubule bundles that extend from the centrosome. The Golgi apparatus is not disrupted by this treatment and remains associated with the microtubule organizing centre (MTOC). This microtubule reorganization has no effect on the ability of CTLs to orient their MTOC towards a bound target cell, nor on their cytolytic activity. In control CTLs, not treated with taxol, a mild hyperthermia treatment (42 degrees C, 30 min) results in an aggregation of the pericentriolar material, a loss of MTOC orientation, an inhibition of cytolytic activity, and a disorganization of the microtubule system [Knox et al.: Exp. Cell Res. 194:275-283, 1991]. In contrast, in taxol-treated CTLs the stabilized microtubule bundles are unaffected by such hyperthermia treatment; however, the other effects of hyperthermia appear identical in control and taxol-treated CTLs. These results indicate that a dynamic, radially arranged microtubule array is not required for the functional polarization of CTLs and suggest that a component of the pericentriolar material may play a key role in effecting MTOC orientation.

Animals↗

PubMed1993

Vitamin E is a complete tumor promoter in mouse skin.

The dorsal skins of 6-8 week old female SENCAR mice were initiated with a single application of 10 nmol of 7,12-dimethylbenz[a]anthracene (DMBA) and subsequently promoted twice/week with topical applications of vitamin E (dl-alpha-tocopherol, 80 mumol/treatment). Vitamin E from two separate commercial suppliers was tested. For comparison, a group of similar mice, also initiated with DMBA, was promoted twice/week with the known tumor promoter 12-O-tetradecanoylphorbol-13- acetate (TPA, 2 micrograms/treatment). Papillomas appeared 39 and 50 days respectively after promotion began with vitamin E from the two different sources, as compared with 32 days in the group receiving TPA promoted. Hundred per cent of TPA-promoted animals and 92-96% of the vitamin E-promoted mice developed tumors. A maximum of 15 papillomas/animal appeared in the TPA-promoted mice. The two vitamin E preparations were somewhat less effective than TPA and showed different relative potencies, producing about seven and 12 papillomas per animal respectively. Unlike TPA, vitamin E showed very little ability to produce an inflammatory response in skin. To test whether initiated cells that did not appear as papillomas after vitamin E promotion were still viable, and had proceeded past stage I of promotion (conversion), the group that developed 12 papillomas/animal from vitamin E promotion was further promoted with mezerein, a stage II promoter. In this group, the papilloma frequency then increased to approximately 17/animal. The animals were followed over the course of their lifespan and monitored for skin carcinomas. In the TPA-promoted group 64% of the mice developed carcinomas, while the two vitamin E-promoted groups showed 48 and 60% incidence respectively. These results indicate that topically applied vitamin E acts as a complete tumor promoter in DMBA-initiated mouse skin, with an efficiency approaching that of TPA. Since vitamin E is a powerful antioxidant, they also suggest that reduction of cellular oxidant levels may trigger the tumor promotional process, and it may therefore be prudent to avoid repetitive or prolonged topical exposure of human skin to antioxidants like vitamin E.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1993

DNA repair in Chlamydomonas reinhardtii induced by heat shock and gamma radiation.

Saccharomyces cerevisiae and Chlamydomonas reinhardtii respond to a sublethal exposure of ionizing radiation by increasing their resistance to killing by a second exposure. We demonstrate here that the two lower eukaryotes apparently achieve this by different mechanisms. We have shown that induced radioresistance in yeast results from increased capacity for recombinational repair, which we believe to occur in G2-phase haploid cells by recombination between homologous chromosomes. This is not possible in G1-phase haploid cells, which lack a second copy of DNA. Haploid C. reinhardtii cells, however, show induced resistance when irradiated asynchronously or in the G1 phase of the cell cycle. We have shown previously that the development of radiation resistance in yeast is proportional to the magnitude of the inducing dose and clearly demonstrates an oxygen effect. There was no oxygen effect for induced radiation resistance in C. reinhardtii cells, but induction remained proportional to dose. In yeast we have reported that both increased radioresistance and thermotolerance are inducible by a heat shock. Here, C. reinhardtii showed induced thermotolerance but no induced radioresistance in response to a heat stress. We have also determined previously that the induced recombinational DNA repair system in yeast recognizes alkylation lesions and therefore confers increased resistance to mutation by MNNG. In these experiments, C. reinhardtii induced for radioresistance were not more resistant to MNNG mutagenesis. These data indicate that haploid C. reinhardtii has a unique DSB repair mechanism. We propose that one possible mechanism may involve chloroplast DNA in a cooperative chloroplast/nuclear recombinational repair process.

Animals↗

PubMed1992

Modification of tumor promotion in the mouse skin by exposure to an alternating magnetic field.

Some epidemiological studies have suggested that exposure to an alternating magnetic field may increase the incidence of some cancers. Our earlier study of carcinogenesis in mouse skin, indicated that exposure to a magnetic field (MF) alone did not promote the growth of tumors. In the present experiment, the ability of a MF to act as a tumor copromoter was investigated. The dorsal skins of female SENCAR mice (6-7-weeks-old) were treated with 10 nmol of 7,12-dimethylbenzanthracene (DMBA) to initiate the carcinogenic process and then tumor development was promoted, for 23 weeks, by weekly applications of 4.9 nmol (0.3 microgram) of 12-0-tetradecanoylphorbal-13-acetate (TPA). One group of 48 mice were exposed to a 60-Hz magnetic field of 2 mT (20 Gauss) for 6 h/day 5 days/week, while a similar group (48 mice) were sham exposed. After week 12, the percentage of mice with tumors and the mean number of tumors per mouse, were higher for the group exposed to MF. At week 18, for example, where the differences between field and sham groups were statistically significant, the percentage of mice with tumors were, respectively, 25% and 8% (P less than 0.05, Fisher exact) and, the mean yield of tumors 1.9 +/- 0.69 and 0.65 +/- 0.46 (mean +/- S.E.M.) (P less than 0.05, Wilcoxon). At week 23 these differences were no longer statistically significant.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1992

A method for hyperthermic treatment of mouse skin.

The Sencar mouse skin system is a recognized model for tumour initiation, promotion and progression. The current interest in the effect of hyperthermia on this multi-stage tumorigenesis model prompted the need for a technique to accurately heat a section of dorsal skin of a large number of mice for 30 min per heat treatment. In the technique described, experimental groups of 25 female Sencar mice were treated at 7-8 weeks of age under general methoxyflurane anaesthesia. Treatment consisted of the application of initiating and/or promoting agents with or without hyperthermia. For hyperthermic skin treatments, each group of mice was placed onto a platform in a water bath so that the dorsal skin of the mice was in contact with 44 degrees C temperature controlled water.

Anesthesia, General↗

PubMed1992

Chronic exposure to ionizing radiation as a tumor promoter in mouse skin.

We have tested chronic exposure to 90Y beta radiation for its action as a complete tumor promoter, a stage I tumor promoter, or a stage II tumor promoter in SENCAR mouse skin. In skin initiated with a single application of 7,12,dimethylbenz[a]anthracene (DMBA, 10 nmol), chronic exposure to beta radiation as a complete promoter (0.5 Gy, twice/week, 13 weeks) produced no tumors and, when added to a complete chemical promoter (TPA), reduced tumor frequency about 30%. A similar result was observed when beta radiation was tested as a stage II promoter. DMBA-initiated mice that received chemical (12-O-tetradecanoylphorbol-13-acetate, TPA) stage I promotion followed by 13 weeks of beta-radiation exposure (0.5 Gy, twice/week) as stage II promotion produced essentially no tumors, and combining the same chronic beta-radiation exposure with chemical (mezerein) stage II promotion reduced tumor frequency about 20% when compared to a similar group that was not irradiated. Chronic beta-radiation exposure was tested two ways as a stage I tumor promoter in initiated skin that was subsequently treated with mezerein as a stage II promoter. Stage I promotion was shown to proceed with the passage of time, indicating this process occurs naturally in the absence of chemical or physical stimulation. Hyperthermia, previously shown to be a potent inhibitor of chemically stimulated stage I promotion, had no effect on the natural process, indicating at least some differences in mechanism between the two processes. The natural process was, in fact, inhibited by chemical tumor promoters, but not by radiation. In addition to the increase resulting from this natural process, tumor frequency was further increased slightly but significantly (12-15%, P less than or equal to 0.05) when chronic radiation exposure was given as a stage I promoter (0.5 Gy, twice/week, 13 weeks) subsequent to initiation, in spite of the expected 20% reduction resulting from this dose. Exposure of initiated animals to radiation (0.5 or 1.0 Gy, twice/week, 2 weeks) in addition to TPA as stage I promotion produced a similar increase in tumor frequency (P less than 0.02). At higher radiation doses, however, tumor frequency was reduced compared to unirradiated controls. In a third test as a stage I promoter, beta radiation (0.5 Gy twice/week, 4 weeks) was given prior to initiation with N-methyl-N'-nitro-N-nitrosoguanidine in animals subsequently promoted by TPA (twice/week, 13 weeks), and again the radiation slightly but significantly (P less than 0.03) increased tumor frequency compared to the unirradiated control group.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1991

Cancer promotion in a mouse-skin model by a 60-Hz magnetic field: II. Tumor development and immune response.

This paper describes preliminary findings on the influence of 60-Hz (2-mT) magnetic fields on tumor promotion and co-promotion in the skins of mice. The effect of magnetic fields on natural killer (NK) cell activity in spleen and blood was also examined. Groups of 32 juvenile female mice were exposed to the magnetic field as described in part I. The dorsal skin of all animals was treated with a subthreshold dose of the carcinogen 7,12-dimethyl-benz(a)anthracene (DMBA). One week after the treatment, two groups were sham exposed (group A) or field exposed at 2 mT (group B) 6 h/day for 21 weeks, to test whether the field would act as a tumor promoter. No tumors developed in these two groups of mice. To test whether the magnetic field would modify tumor development by directly affecting tumor growth or by suppressing immune surveillance, two additional groups of mice were treated weekly with the tumor promoter 12-0-tetradecanoylphorbol-13-acetate (TPA) and then either sham exposed (group C) or field exposed (group D). The time to appearance of tumors was shorter (but not statistically so) in the group exposed to magnetic fields and TPA. Some differences in NK cell activity and spleen size were observed between the sham- and field-exposed groups.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1991

Effects of hyperthermia on microtubule organization and cytolytic activity of murine cytotoxic T lymphocytes.

When murine cytotoxic T lymphocytes (CTL) are heated at 42 degrees C for 30 min their ability to lyse their target cells (TC) is severely impaired. When the CTL are allowed to recover at 37 degrees C, a partial recovery of cytolytic activity that peaks within 6 h is observed. A dye exclusion assay demonstrated that such a heat shock does not affect the viability of the CTL and direct microscopic observations established that their ability to bind to TC is not impaired. Therefore, the step or steps inhibited by hyperthermia are subsequent to TC recognition and binding. Kupfer et al. ((1983) Proc. Natl. Acad. Sci. USA 80, 7224-7228) demonstrated that upon binding to an appropriate TC, a rapid orientation of the Golgi apparatus and the microtubule organizing center (MTOC) occurred within the CTL so that the two organelles face the TC. This orientation is a prerequisite for efficient TC lysis. We have shown by immunofluorescence and confocal microscopy, using a monoclonal antibody to tubulin and a rabbit autoimmune serum that binds a centriole-associated protein, that the organization of the MTOC-microtubule array is disrupted by hyperthermia. EM suggests that this disorganization of the microtubules may result from an aggregation of the pericentriolar material. The recovery of cytolytic activity is coincident with the reorganization of the microtubules about the MTOC. These findings suggest that the initial inhibitory effect of hyperthermia on CTL function results from the disruption of microtubule organization.

Animals↗

PubMed1991

DNA lesions that signal the induction of radioresistance and DNA repair in yeast.

DNA recombinational repair, and an increase in its capacity induced by DNA damage, is believed to be the major mechanism that confers resistance to killing by ionizing radiation in yeast. We have examined the nature of the DNA lesions generated by ionizing radiation that induce this mechanism, using two different end points: resistance to cell killing and ability of the error-free recombinational repair system to compete for other DNA lesions and thereby suppress chemical mutation. Under the various conditions examined in this study, the "maximum" inducible radiation resistance was increased approximately 1.5- to 3-fold and suppression of mutation about 10-fold. DNA lesions produced by low-LET gamma rays at doses greater than about 20 Gy given in oxygen were shown to be more efficient, per unit dose, at inducing radioresistance to killing than were lesions produced by neutrons (high-LET radiation). This suggests that DNA single-strand breaks are more important lesions in the induction of radioresistance than DNA double-strand breaks. Oxygen-modified lesions produced by gamma rays (low-LET radiation) were particularly efficient as induction signals. DNA damage due to hydroxyl radicals (OH.) derived from the radiolytic decomposition of H2O produced lesions that strongly induced this DNA repair mechanism. Similarly, OH. derived from aqueous electrons (e-aq) in the presence of N2O also efficiently induced the response. Cells induced to radioresistance to killing with high-LET radiation did not suppress N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-generated mutations as well as cells induced with low-LET radiation, supporting the conclusion that the type of DNA damage produced by low-LET radiation is a better inducer of recombinational repair. Surprisingly, however, cells induced with gamma radiation in the presence of N2O that became radioresistant to killing were unable to suppress MNNG mutations. This result indicates that OH. generated via e-aq (in N2O) may produce unusual DNA lesions which retard normal repair and render the system unavailable to compete for MNNG-generated lesions. We suggest that the repairability of these unique lesions is restricted by either their chemical nature or topological accessibility. Attempted repair of these lesions has lethal consequences and accounts for N2O radiosensitization of repair-competent but not incompetent cells. We conclude that induction of radioresistance in yeast by ionizing radiation responds variably to different DNA lesions, and these affect the availability of the induced recombinational repair system to deal with subsequent damage.

Cell Survival↗

PubMed1990

The involvement of topoisomerases and DNA polymerase I in the mechanism of induced thermal and radiation resistance in yeast.

Either an ionizing radiation exposure or a heat shock is capable of inducing both thermal tolerance and radiation resistance in yeast. Yeast mutants, deficient in topoisomerase I, in topoisomerase II, or in DNA polymerase I, were used to investigate the mechanism of these inducible resistances. The absence of either or both topoisomerase activities did not prevent induction of either heat or radiation resistance. However, if both topoisomerase I and II activities were absent, the sensitivity of yeast to become thermally tolerant (in response to a heat stress) was markedly increased. The absence of only topoisomerase I activity (top1) resulted in the constitutive expression of increased radiation resistance equivalent to that induced by a heat shock in wild-type cells, and the topoisomerase I-deficient cells were not further inducible by heat. This heat-inducible component of radiation resistance (or its equivalent constitutive expression in top1 cells) was, in turn, only a portion of the full response inducible by radiation. The absence of polymerase I activity had no detectable effect on either response. Our results indicate that the actual systems that confer resistance to heat or radiation are independent of either topoisomerase activity or DNA polymerase function, but suggest that topoisomerases may have a regulatory role during the signaling of these mechanisms. The results of our experiments imply that maintenance of correct DNA topology prevents induction of the heat-shock response, and that heat-shock induction of a component of the full radiation resistance in yeast may be the consequence of topoisomerase I inactivation.

Adaptation, Physiological↗

PubMed1990

Beta-radiation-induced resistance to MNNG initiation of papilloma but not carcinoma formation in mouse skin.

We have shown previously that the risk of tumor initiation, promotion, and progression in animals initiated with alkylating agents can be drastically altered by hyperthermia treatments. We show here that ionizing radiation can also alter the risk of tumor initiation by alkylating agents. Using a two-step skin tumorigenesis protocol in female SENCAR mice (initiation by MNNG, promotion with TPA), we exposed the dorsal skin of the mice to various doses of 90Sr/90Y beta radiation near the time of initiation. The radiation produced a dose-dependent reduction in the number of papillomas which appeared after TPA promotion, with about a 20% reduction in animals receiving 0.5 Gy surface dose just before initiation, about 50% reduction after 2.5 Gy, and greater than 80% at doses above 5 Gy. A dose of 2.5 Gy in animals initiated with DMBA produced no significant reduction. One skin hyperthermia treatment (44 degrees C, 30 min) along with radiation in MNNG-initiated animals partially blocked the protective effect of radiation and increased the papilloma frequency. Radiation (2.5 Gy) given either 6 days before or after MNNG initiation was less effective but still reduced papilloma frequency about 20%. In sharp contrast to the marked reduction in papilloma formation, these same animals showed no change in carcinoma frequency with any of the doses or schedules of beta radiation. MNNG initiation alone produced three types of initiated cells. One type, produced in low yield, was promotion-independent with a high probability of progression to a carcinoma and appeared unaffected by the radiation. A second type, produced in intermediate yield, was promotion-dependent and also had a high progression probability, but was likewise unaffected by the radiation. The third and most abundant type was promotion-dependent with a very low progression probability. Radiation exposure resulted in a decrease in the risk of an MNNG initiation event which led only to the third type of cell. The data therefore indicate that the risk of some, but not all, tumor-initiating events caused by alkylating agents can be reduced by an exposure to ionizing radiation.

Animals↗

PubMed1988

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)

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1987

Inducible DNA-repair systems in yeast: competition for lesions.

DNA lesions may be recognized and repaired by more than one DNA-repair process. If two repair systems with different error frequencies have overlapping lesion specificity and one or both is inducible, the resulting variable competition for the lesions can change the biological consequences of these lesions. This concept was demonstrated by observing mutation in yeast cells (Saccharomyces cerevisiae) exposed to combinations of mutagens under conditions which influenced the induction of error-free recombinational repair or error-prone repair. Total mutation frequency was reduced in a manner proportional to the dose of 60Co-gamma- or 254 nm UV radiation delivered prior to or subsequent to an MNNG exposure. Suppression was greater per unit radiation dose in cells gamma-irradiated in O2 as compared to N2. A rad3 (excision-repair) mutant gave results similar to wild-type but mutation in a rad52 (rec-) mutant exposed to MNNG was not suppressed by radiation. Protein-synthesis inhibition with heat shock or cycloheximide indicated that it was the mutation due to MNNG and not that due to radiation which had changed. These results indicate that MNNG lesions are recognized by both the recombinational repair system and the inducible error-prone system, but that gamma-radiation induction of error-free recombinational repair resulted in increased competition for the lesions, thereby reducing mutation. Similarly, gamma-radiation exposure resulted in a radiation dose-dependent reduction in mutation due to MNU, EMS, ENU and 8-MOP + UVA, but no reduction in mutation due to MMS. These results suggest that the number of mutational MMS lesions recognizable by the recombinational repair system must be very small relative to those produced by the other agents. MNNG induction of the inducible error-prone systems however, did not alter mutation frequencies due to ENU or MMS exposure but, in contrast to radiation, increased the mutagenic effectiveness of EMS. These experiments demonstrate that in this lower eukaryote, mutagen exposure does not necessarily result in a fixed risk of mutation, but that the risk can be markedly influenced by a variety of external stimuli including heat shock or exposure to other mutagens.

DNA Damage↗

PubMed1987

Tumorigenesis and carcinogenesis in mouse skin treated with hyperthermia during stage I or stage II of tumor promotion.

The action of hyperthermia treatments on tumor promotion separated into a two-stage protocol has been investigated. 7,12-Dimethylbenz[a]anthracene (DMBA) initiated dorsal skin of female SENCAR mice was promoted with either H2O2 or 12-O-tetradecanoylphorbol-13-acetate (TPA) (4 applications, 2 times/week) as the first stage of promotion, followed by promotion with mezerein (28 applications, 2 times/week) as the second stage. Hyperthermia (44 degrees C, 30 min) treatment of the skin at the time of stage II promotion only (just before each mezerein application) suppressed 100% of papillomas when H2O2 was used as a first-stage promoter and 96% when TPA was used as the first stage, as compared to unheated control animals. The same hyperthermia treatment given only at stage I of promotion had similar results. Hyperthermia treatments just before stage I TPA promotion (4 treatments only) followed by mezerein as the second stage reduced papilloma formation by 92%. When H2O2 was used as the first stage promoter and again mezerein as the second, papilloma frequency was reduced by 74%, as compared to unheated controls. This antipromotion activity of hyperthermia could not be linked to an inhibition of skin protease activity. Although papilloma frequency was markedly suppressed by hyperthermia during stage I promotion only, carcinoma formation was not. A similar number of carcinomas appeared in the groups of mice receiving hyperthermia with either H2O2 or TPA as first-stage promoters, as in comparable groups receiving no hyperthermia. In contrast, when hyperthermia treatments were given during stage II promotion with mezerein (using either H2O2 or TPA as stage I promoters), carcinomas (as well as papillomas) were markedly reduced. The results suggest that DMBA initiation creates two types of promotion-dependent cells, a majority with relatively low progression probability and a minority with relatively high progression probability. The former require both stage I and II promotion while the latter require only stage II promotion to form tumors. Hyperthermia treatments given during stage II promotion protected against promotion and progression of both types of initiated cells, but similar treatments only during stage I did not protect against promotion and progression of the latter. Although promotion was required for expression, relative progression probability appeared linked to initiation and not promotion events. These findings suggest that hyperthermia treatment of persons exposed to tumor-promoting agents may reduce the risk of induced tumorigenesis.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1986

Hyperthermia and phorbol ester tumor promotion in mouse skin.

In a two-stage skin tumorigenesis protocol [7,12-dimethylbenz[a]anthracene (DMBA) initiation followed by twice weekly 12-O-tetradecanoylphorbol-13-acetate (TPA) promotion], SENCAR mice developed an average of approximately 8.5 papillomas per animal. Hyperthermia treatments of the initiated skin (44 degrees C, 30 min) immediately before or after each TPA application (for 90 days) reduced papilloma frequency 80-90%. Animals whose initiated skin was made thermo-tolerant at the time of TPA application (by hyperthermia treatment 24 h prior to each application of promoter) showed slightly less protection (approximately 70% reduction in frequency). Multiple 44 degrees C hyperthermia treatments alone (27 X, twice a week) had no promoting activity in DMBA-initiated skin. The usual responses of skin to TPA promotion, including an increase in dark cells, epidermal thickening, reddening and erosion were all suppressed in animals treated with hyperthermia near the time of TPA application. The effect of hyperthermia on tumorigenesis was at the promotion stage and the survival of initiated cells was not affected, since the normal number of papillomas was produced when TPA promotion was delayed until after the multiple (27 X, twice a week) hyperthermia treatments were completed. Hyperthermia treatments (44 degrees C, 30 min, twice weekly for 90 days) given near the time of TPA application also suppressed the incidence of carcinomas appearing within 300 days. About 40% of the DMBA-initiated, TPA-promoted animals developed a carcinoma, compared with only approximately 10% of a similar group which received hyperthermia treatments near each TPA application. Papillomas appearing in spite of hyperthermia treatments during promotion were not more likely to progress into carcinomas than those appearing in unheated animals. Such hyperthermia treatments given to animals bearing pre-existing papillomas did not markedly alter the subsequent development of carcinomas compared with unheated controls. The results demonstrated that 44 degrees C hyperthermia applied near the time of TPA promotion acted as a powerful antipromoter and suppressed the appearance of both papillomas and carcinomas, apparently by acting at an early stage of promotion.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1985

Triggering of DNA strand breaks by 45 degrees C hyperthermia and its influence on the repair of gamma-radiation damage in human white blood cells.

Human peripheral white blood cells, freshly isolated from normal individual donors, were exposed to hyperthermia. Heat-generated DNA strand break damage and white blood cell capacity to repair radiation-induced breaks were determined by a fluorometric alkaline unwinding assay. Strand breaks could be readily detected when white blood cells were incubated in a physiological salt solution at temperatures between 41 degrees and 46 degrees C, for times up to 90 min. The time course of strand break induction at 45 degrees C was characterized by a short initial lag, followed by a period of rapid break induction and subsequently a lower rate. Evidence is presented which suggests that the induction of DNA damage involved a "triggering" mechanism; a short treatment at 45 degrees C (10 to 20 min) initiated a cellular event which led to a rapid increase in the number of strand breaks during subsequent incubation of 37 degrees C. Continuous incubation at 45 degrees C produced less DNA damage than an initial period at 45 degrees C followed by incubation at 37 degrees C. This apparent "triggering phenomenon" was not due to a triggering of the respiratory burst in phagocytic cells, since no O2- could be detected; in fact, a 30-min treatment at 45 degrees C largely blocked the capacity of the cells to respond normally to a soluble stimulator of the respiratory burst. Unlike gamma-ray-induced breaks, 45 degrees C hyperthermia-induced breaks did not rejoin during subsequent incubation for up to 1 h at 37 degrees C. Additionally, 45 degrees C hyperthermia treatment progressively inhibited the ability of the cells to repair subsequent gamma-ray-induced breaks (4 Gy). This inhibition occurred during the period in which 45 degrees C heat rapidly induced strand breaks. Hyperthermia (41 degrees C), which did not trigger strand breaks, did not cause detectable inhibition of this repair capacity. There was no indication that hyperthermia sensitized cells to radiation-induced strand breaks.

Cell Survival↗

PubMed1985

Sensitivity of hyperthermia-treated human cells to killing by ultraviolet or gamma radiation.

Human xeroderma pigmentosum (XP) or Fanconi anemia (FA) fibroblasts displayed shouldered 45 degrees C heat survival curves not significantly different from normal fibroblasts, a result similar to that previously found for ataxia telangiectasia (AT) cells, indicating heat resistance is not linked to either uv or low-LET ionizing radiation resistance. Hyperthermia (45 degrees C) sensitized normal and XP fibroblasts to killing by gamma radiation but failed to sensitize the cells to the lethal effects of 254 nm uv radiation. Thermal inhibition of repair of ionizing radiation lesions but not uv-induced lesions appears to contribute synergistically to cell death. The thermal enhancement ratio (TER) for the synergistic interaction of hyperthermia (45 degrees C, 30 min) and gamma radiation was significantly lower in one FA and two strains (TER = 1.7-1.8) than that reported previously for three normal strains (TER = 2.5-3.0). These XP and FA strains may be more gamma sensitive than normal human fibroblasts. Since hyperthermia treatment only slightly increases the gamma-radiation sensitivity of ataxia telangiectasia (AT) fibroblasts compared to normal strains, it is possible that the degree of thermal enhancement attainable reflects the genetically inherent ionizing radiation repair capacity of the cells. The data indicate that both repair inhibition and particular lesion types are required for lethal synergism between heat and radiation. We therefore postulate that the transient thermal inhibition of repair results in the conversion of gamma-induced lesions to irrepairable lethal damage, while uv-type damage can remain unaltered during this period.

Cell Survival↗