ABSTRACT
SGI-110 is a second-generation hypomethylating prodrug whose active metabolite is the well-characterized drug decitabine. This novel compound is an oligonucleotide consisting of decitabine linked through a phosphodiester bond to the endogenous nucleoside deoxyguanosine. The dinucleotide configuration provides protection from drug clearance by deamination, while maintaining at least equivalent effects on gene-specific and global hypomethylation both in vitro and in animal model systems. This agent is currently being tested in phase I and II clinical trials in humans and has been demonstrated to be safe and well tolerated as a single agent, with evidence of promising activity in heavily pretreated (including currently FDA approved hypomethylating drugs) myelodysplastic syndrome and acute myeloid leukemia patients. Ongoing trials are also open in platinum-resistant ovarian cancer and hepatocellular carcinoma.
ABSTRACT
BACKGROUND: Upregulation of PIM kinase expression has been reported in many malignancies, suggesting that inhibition of PIM kinase activity may be an attractive therapeutic strategy. We hypothesised that inhibition of PIM kinase activity with SGI-1776, a novel small molecule inhibitor of PIM kinase activity, would reduce the viability of renal cell carcinoma (RCC) cells and enhance the activity of sunitinib. METHODS: Immunoblotting, qRT-PCR, and gene expression arrays were carried out to identify genes modulated by SGI-1776 treatment. The anticancer activity of SGI-1776 and sunitinib was determined by viability and apoptosis assays and in tumour xenografts in vivo. RESULTS: Treatment with SGI-1776 led to a decrease in phosphorylated and total c-Myc levels, which resulted in the modulation of c-Myc target genes. SGI-1776 in combination with sunitinib induced a further reduction in c-Myc levels, which was associated with enhanced anticancer activity. siRNA-mediated knockdown of c-Myc demonstrated that its expression has a key role in regulating the sensitivity to the combination of SGI-1776 and sunitinib. Importantly, the combination significantly reduced tumour burden in two RCC xenograft models compared with single-agent therapy and was very well tolerated. CONCLUSION: These data indicate that targeting PIM kinase signalling is a promising treatment strategy for RCC.
Subject(s)
Antineoplastic Agents/pharmacology , Carcinoma, Renal Cell/pathology , Imidazoles/pharmacology , Indoles/pharmacology , Kidney Neoplasms/pathology , Protein Kinase Inhibitors/pharmacology , Proto-Oncogene Proteins c-pim-1/antagonists & inhibitors , Pyridazines/pharmacology , Pyrroles/pharmacology , Animals , Carcinoma, Renal Cell/enzymology , Cell Line, Tumor , Female , Humans , Immunohistochemistry , Kidney Neoplasms/enzymology , Mice , Mice, Inbred BALB C , Mice, Nude , Phosphorylation , Proto-Oncogene Proteins c-myc/metabolism , Real-Time Polymerase Chain Reaction , SunitinibABSTRACT
Previously we showed that a mismatch repair (MMR)-deficient cell line, HCT116 (hMLH1 mut), unlike a MMR wild-type cell line, SW480, was more resistant to the therapeutic methylating agent, temozolomide (TMZ), because the MMR complex fails to recognize TMZ-induced O6-methylguanine DNA adduct mispairings with thymine that arise after replication. TMZ also produces N7-methylguanine and N3-methyladenine adducts that are processed efficiently by the base excision repair (BER) system. After removal of the methylated base by methylpurine glycosylase, which creates the abasic or apurinic-apyrimidinic (AP) site, the phosphodiester bond is hydrolyzed immediately by AP endonuclease, initiating the repair of the AP site. Methoxyamine (MX) reacts with the abasic site and prevents AP endonuclease cleavage, disrupting DNA repair. MX potentiated the cytotoxic effect of TMZ with a dose modification factor (DMF) of 2.3+/-0.12 in SW480 and 3.1+/-0.16 in HCT116. When combined with O6-benzylguanine (BG), MX and TMZ dramatically increased TMZ cytotoxicity (65.8-fold) in SW480, whereas no additive effect was seen in HCT116. This suggests that N7-methylguanine and N3-methyladenine adducts are cytotoxic lesions in MMR-deficient and wild-type cells when BER is interrupted. Because poly(ADP-ribose) polymerase (PARP) aids in processing of DNA strand breaks induced during MMR and BER, we asked whether PARP inhibitors would also affect BER-mediated cell killing. We found that PARP inhibitors PD128763, 3-aminobenzimide, and 6-aminonicotinamide increased the sensitivity to TMZ in both HCT116 MMR-deficient cells and SW480 MMR wild-type cells. In HCT116 cells, PD128763 remarkably decreased resistance to TMZ, with a DMF of 4.7+/-0.2. However, the combination of PD128763, BG, and TMZ had no greater effect, indicating that persistent O6-methylguanine had no effect on cytotoxicity. In SW480, the DMF for TMZ cytotoxicity was 3.1+/-0.12 with addition of PD128763 and 36 with addition of PD128763 and BG. Synergy analysis by median effect plots indicated a high degree of synergy between TMZ and MX or PD128763. In contrast, 1,3-bis(2-chloroethyl)-1-nitrosourea combined with either MX or PD128763 showed little if any potentiation observed in the absence of BG in either cell line, suggesting that BER pathway has little impact on cytotoxic processing of 1,3-bis(2-chloroethyl)-1-nitrosourea-induced adducts. These studies indicate that targeting BER with MX or PARP inhibitors enhances the cytotoxicity of methylating agents, even in MMR-deficient cells.
Subject(s)
Antineoplastic Agents, Alkylating/pharmacology , Colonic Neoplasms/drug therapy , DNA Repair/drug effects , Carmustine/pharmacology , Cell Cycle/drug effects , Colonic Neoplasms/genetics , Dacarbazine/analogs & derivatives , Dacarbazine/pharmacology , Drug Synergism , Humans , Hydroxylamines/pharmacology , Isoquinolines/pharmacology , Poly(ADP-ribose) Polymerase Inhibitors , TemozolomideABSTRACT
PURPOSE AND METHODS: The lack of a functional DNA mismatch repair (MMR) pathway has been recognized as a common characteristic of several different types of human cancers due to mutation affecting one of the MMR genes or due to promoter methylation gene silencing. These MMR-deficient cancers are frequently resistant to alkylating agent chemotherapy such as DNA-methylating or platinum-containing compounds. To correlate drug resistance with MMR status in a large panel of human tumor cell lines, we evaluated by Western blot the cellular levels of the two MMR proteins most commonly mutated in human cancers, MLH1 and MSH2, in the NCI human tumor cell line panel. This panel consists of 60 cell lines distributed among nine different neoplastic diseases. RESULTS: We found that in most of these cell lines both MLH1 and MSH2 were expressed, although at variable levels. Five cell lines (leukemia CCRF-CEM, colon HCT 116 and KM12 and ovarian cancers SK-OV-3 and IGROV-1) showed complete deficiency in MLH1 protein. MSH2 protein was detected in all 57 cell lines studied. Absence of MLH1 protein was always linked to resistance to the methylating chemotherapeutic agent temozolomide. This resistance was independent of cellular levels of O6-alkylguanine DNA alkyltransferase. Based on data available for review in the NCI COMPARE database, cellular levels of MLH1 and MSH2 did not correlate significantly with sensitivity to any standard anticancer drug or with any characterized molecular target already tested against the same panel of cell lines. CONCLUSION: Based on evaluation of 60 tumor cell lines in the NCI anticancer drug screen, MLH1 deficiency was more common than MSH2 deficiency and was always associated with a high degree of temozolomide resistance. These data will enable correlations with other drug sensitivities and molecular targets in the COMPARE database to evaluate linked processes in tumor drug resistance.
Subject(s)
DNA Repair , DNA-Binding Proteins , Dacarbazine/analogs & derivatives , Drug Resistance, Neoplasm/genetics , Neoplasm Proteins/metabolism , Proto-Oncogene Proteins/metabolism , Tumor Cells, Cultured/metabolism , Adaptor Proteins, Signal Transducing , Antineoplastic Agents, Alkylating/pharmacology , Blotting, Western , Carrier Proteins , Dacarbazine/pharmacology , Drug Screening Assays, Antitumor , Humans , MutL Protein Homolog 1 , MutS Homolog 2 Protein , Neoplasm Proteins/deficiency , Nuclear Proteins , Proto-Oncogene Proteins/deficiency , Software , Temozolomide , Tumor Cells, Cultured/drug effectsABSTRACT
We investigated the ability of alachlor to cause DNA damage by measuring single-strand breaks (SSB) in DNA, after metabolic activation by freshly isolated rat hepatocytes. Incubation of different concentrations of alachlor with rat hepatocytes led to numerous metabolites. The majority, isolated and identified by GC-MS analysis, were products arising from reactions catalyzed by the P-450 monooxygenase system, arylamidase and flavin mixed-function oxidase/cytochrome P-450 monooxygenase. The results, using freshly isolated rat hepatocytes, showed that in these conditions several potentially DNA damaging metabolites were produced; this experimental condition was used to assess DNA damage induced by the mixture of alachlor and its metabolites. The alkaline elution technique showed that at 200 microM and more clearly at 400 microM there were some small fragments that eluted in the first fraction. This fragmentation was probably due to alachlor cytotoxicity. In addition to the small DNA fragments eluting in the first fraction there were other larger DNA fragments. These DNA-SSB were most evident at the alachlor concentration of 400 microM, but also at 200 microM and 100 microM, whereas at 10 microM the DNA elution rate appeared comparable to that of controls. The results suggest that some unstable and DNA-reactive metabolites might interact with DNA causing SSB and such interaction might be important in relation to the mechanism of alachlor-induced DNA damage. However, it may not be possible to clarify whether SSB are the result of direct DNA interaction of the compound or of secondary cellular processes after chemical treatment.
Subject(s)
Acetamides/toxicity , DNA Damage , Herbicides/toxicity , Liver/drug effects , Acetamides/metabolism , Animals , Biotransformation , Cell Survival/drug effects , Cells, Cultured , DNA, Single-Stranded/metabolism , Gas Chromatography-Mass Spectrometry , Herbicides/metabolism , Liver/metabolism , Male , Mice , RatsABSTRACT
Using L1210 and a subline resistance to chloroethylnitrosoureas (L1210/BCNU), we found that the resistance to 1-(2-chloroethyl)-1-nitrosourea (CNU) or to diethyl-1-3-(2-chloroethyl)-3-nitrosoureido ethyl phosphonate (fotemustine) can be reversed by a pretreatment with O6-methyl Guanine (O6-mGua) or temozolomide. In L1210/BCNU but not in L1210 the pretreatment with O6mGua caused an increased peak level of CNU-induced DNA-interstrand crosslinks. We then evaluated whether the resistance to BCNU could be counteracted in vivo by i.p. O6mGua treatment of L1210/BCNU bearing mice. The results were negative due to the fact that O6mGua, which was not toxic when given alone, caused a high toxicity when associated with BCNU.
Subject(s)
Antineoplastic Agents/pharmacology , Carmustine/pharmacology , Dacarbazine/analogs & derivatives , Ethylnitrosourea/analogs & derivatives , Guanine/analogs & derivatives , Leukemia L1210/drug therapy , Nitrosourea Compounds/pharmacology , Organophosphorus Compounds/pharmacology , Animals , Carmustine/therapeutic use , Cell Cycle/drug effects , Cell Division/drug effects , Dacarbazine/pharmacology , Dacarbazine/therapeutic use , Drug Evaluation, Preclinical , Drug Resistance , Ethylnitrosourea/pharmacology , Ethylnitrosourea/therapeutic use , Guanine/pharmacology , Guanine/therapeutic use , Kinetics , Mice , Mice, Inbred DBA , Nitrosourea Compounds/therapeutic use , Organophosphorus Compounds/therapeutic use , TemozolomideABSTRACT
We have previously shown that human cancer cells deficient in DNA mismatch repair (MMR) are resistant to the chemotherapeutic methylating agent temozolomide (TMZ) and can be sensitized by the base excision repair (BER) blocking agent methoxyamine (MX) [21]. To further characterize BER-mediated repair responses to methylating agent-induced DNA damage, we have now evaluated the effect of MX on TMZ-induced DNA single strand breaks (SSB) by alkaline elution and DNA double strand breaks (DSB) by pulsed field gel electrophoresis in SW480 (O6-alkylguanine-DNA-alkyltransferase [AGT]+, MMR wild type) and HCT116 (AGT+, MMR deficient) colon cancer cells. SSB were evident in both cell lines after a 2-h exposure to equitoxic doses of temozolomide. MX significantly increased the number of TMZ-induced DNA-SSB in both cell lines. In contrast to SSB, TMZ-induced DNA-DSB were dependent on MMR status and were time-dependent. Levels of 50 kb double stranded DNA fragments in MMR proficient cells were increased after TMZ alone or in combination with O6-benzylguanine or MX, whereas, in MMR deficient HCT116 cells, only TMZ plus MX produced significant levels of DNA-DSB. Levels of AP endonuclease, XRCC1 and polymerase beta were present in both cell lines and were not significantly altered after MX and TMZ. However, cleavage of a 30-mer double strand substrate by SW480 and HCT116 crude cell extracts was inhibited by MX plus TMZ. Thus, MX potentiation of TMZ cytotoxicity may be explained by the persistence of apurinic/apyrimidinic (AP) sites not further processed due to the presence of MX. Furthermore, in MMR-deficient, TMZ-resistant HCT116 colon cancer cells, MX potentiates TMZ cytotoxicity through formation of large DS-DNA fragmentation and subsequent apoptotic signalling.
Subject(s)
Colonic Neoplasms/genetics , DNA Damage , DNA, Neoplasm/drug effects , Dacarbazine/pharmacology , Hydroxylamines/pharmacology , Apoptosis/drug effects , Blotting, Western , Colonic Neoplasms/pathology , DNA Repair , Dacarbazine/analogs & derivatives , Drug Synergism , Electrophoresis, Gel, Pulsed-Field , Humans , In Situ Nick-End Labeling , Temozolomide , Tumor Cells, CulturedABSTRACT
In mammalian cells, under aerobic conditions, ionizing radiations and radiomimetic chemical agents can induce an enzymatic activity involved in DNA repair, O6-alkylguanine-DNA alkyltransferase (O6-AT). This catalytic protein is active against alkyl-radical-induced DNA damages. This induction was proposed to be linked to the formation of hydroxyl radicals. The possible involvement of O6-AT in the defense mechanism of the cell against aerobic radiation damage was investigated by comparing the X-ray sensitivity of two Chinese hamster ovary (CHO) cell lines, the first deficient (CHO mex-) and the second proficient by transfection of O6-AT (CHO mex+). The colony-forming ability after X-irradiation was appreciably reduced in CHO mex- in comparison to CHO mex+ cells. Nevertheless, pretreatment of proficient cells with O6-methylguanine, a specific inhibitor of O6-AT, reduced the DNA repair activity but did not modify the degree of sensitivity to X-rays of the CHO mex+ cells. Since the glutathione concentrations as well as the DNA damage amounts induced by X-irradiation were comparable in the variously treated cell lines, these results suggest that the observed induction of O6-AT by ionizing radiation in aerobic conditions could be a generalized rather than a specific response to damage by radicals.
Subject(s)
CHO Cells/radiation effects , DNA Damage , Methyltransferases/physiology , Radiation Tolerance , Alkylation , Animals , CHO Cells/enzymology , Cricetinae , Cricetulus , DNA Repair , Free Radical Scavengers , Glutathione/analysis , Methyltransferases/deficiency , Methyltransferases/metabolism , O(6)-Methylguanine-DNA MethyltransferaseABSTRACT
Twenty-five patients suffering from chronic renal insufficiency and who had been undergoing dialysis for a long time were tested on the Wechsler-Bellevue Intelligence Scale with a control of twenty-five subjects of the same age with a comparable educational and social background. The results of the statistical analysis of the difference between the two groups demonstrates a significant psychological deterioration in the group of uremic patients which was particularly noticeable in the performance test. The rate of deterioration was not significantly related to the duration of the dialysis. At the same time electromyography (motor conduction velocity of the S.P.E. nerve) show in 40% of the uremic patients evidence of neuropathy.
Subject(s)
Electromyography , Kidney Failure, Chronic/psychology , Renal Dialysis , Adult , Female , Humans , Intellectual Disability/diagnosis , Intelligence Tests , Kidney Failure, Chronic/therapy , Male , Middle Aged , Neural ConductionSubject(s)
Aminobutyrates/therapeutic use , Glutamine/therapeutic use , Intellectual Disability/drug therapy , Mental Processes/drug effects , Adolescent , Aminobutyrates/pharmacology , Brain/drug effects , Child , Drug Combinations , Electroencephalography , Evaluation Studies as Topic , Glutamine/pharmacology , Humans , Memory/drug effects , Psychological TestsSubject(s)
Anxiety/drug therapy , Depression/drug therapy , Hypochondriasis/drug therapy , Mental Disorders/drug therapy , Morpholines/therapeutic use , Sleep Initiation and Maintenance Disorders/drug therapy , Tranquilizing Agents/therapeutic use , Adolescent , Adult , Aged , Female , Humans , Male , Middle AgedSubject(s)
Anemia, Macrocytic/pathology , Duodenum/pathology , Intestinal Mucosa/pathology , Jejunum/pathology , Mast Cells , Adult , Aged , Female , Humans , Male , Middle AgedABSTRACT
Escherichia coli ada ogt mutants, which are totally deficient in O6-methylguanine-DNA methyltransferases, have an increased spontaneous mutation rate. This phenotype is particularly evident in starving cells and suggests the generation of an endogenous DNA alkylating agent under this growth condition. We have found that in wild-type cells, the level of the inducible Ada protein is 20-fold higher in stationary-phase and starving cells than in rapidly growing cells, thus enhancing the defense of these cells against DNA damage. The increased level of Ada in stationary cells is dependent on RpoS, a stationary-phase-specific sigma subunit of RNA polymerase. We have also identified a potential source of the mutagenic agent. Nitrosation of amides and related compounds can generate directly acting methylating agents and can be catalyzed by bacteria] enzymes. E. coli moa mutants, which are defective in the synthesis of a molybdopterin cofactor required by several reductases, are deficient in nitrosation activity. It is reported here that a moa mutant shows reduced generation of a mutagenic methylating agent from methylamine (or methylurea) and nitrite added to agar plates. Moreover, a moa mutation eliminates much of the spontaneous mutagenesis in ada ogt mutants. These observations indicate that the major endogenous mutagen is not S-adenosylmethionine but arises by bacterially catalyzed nitrosation.
Subject(s)
Bacterial Proteins/metabolism , DNA, Bacterial/metabolism , Escherichia coli Proteins , Escherichia coli/metabolism , Methyltransferases/metabolism , Nitrogen/metabolism , DNA Damage , DNA Methylation , Escherichia coli/genetics , Methyltransferases/genetics , Mutation , O(6)-Methylguanine-DNA Methyltransferase , Transcription FactorsABSTRACT
Temozolomide is a new anticancer agent which in the early clinical investigation has shown promising antitumor activity. It decomposes spontaneously to the active metabolite of DTIC (MTIC). Temozolomide is more cytotoxic against L1210 than against a subline L1210/BCNU, resistant to chloroethylnitrosoureas. Using [methyl-3H] temozolomide we found that after 1 h exposure the amount of O6-methylguanine (O6mGua) was twice as high in L1210 than in L1210/BCNU whereas the amount of N7 mGua was approximately the same in the two cell lines. O6-alkylguanine DNA alkyltransferase (AT) levels were higher in L1210/BCNU than in L1210, supporting the view that the resistance to methyltriazenes is probably related to the efficient repair of O6mGua in L1210/BCNU. Exposure of L1210/BCNU cells to 0.4 mM O6mGua for 24 h resulted in a depletion of AT and in a higher temozolomide-induced cytotoxicity. In the sensitive cell line L1210, temozolomide activity was not potentiated by O6mGua pretreatment. Moreover, in L1210/BCNU, O6mGua increased DNA single-strand breaks caused by temozolomide, suggesting that O6-guanine alkylation induces an excision repair mechanism in cells depleted in AT.
Subject(s)
Antineoplastic Agents/pharmacology , DNA Damage/drug effects , DNA, Neoplasm/drug effects , Dacarbazine/analogs & derivatives , Guanine/analogs & derivatives , Leukemia L1210/drug therapy , Alkalies , Alkylation , Animals , Cell Division/drug effects , Dacarbazine/pharmacology , Drug Resistance/physiology , Drug Screening Assays, Antitumor , Guanine/pharmacology , Methyltransferases/metabolism , Mice , Nitrosourea Compounds/pharmacology , O(6)-Methylguanine-DNA Methyltransferase , Temozolomide , Tumor Cells, CulturedABSTRACT
Previous studies have indicated that excision repair genes, such as ERCC1, or early response genes, such as c-fos, may play a significant role in regulating cellular responses to cisplatin (CDDP) by mediating DNA synthesis and repair pathways. This present study aimed to determine whether altered gene expression mediated CDDP resistance expressed in two human tumour sublines following their in vitro exposure to fractionated X-irradiation, not to the drug itself. These sublines, designated SuSa/DXR10 and SKOV-3/DXR10, established respectively from a testicular teratoma cell line (SuSa) or an ovarian carcinoma cell line (SKOV-3), expressed stable 3.1- and 2-fold levels of CDDP resistance, as judged by clonogenic assay. Both sublines expressed c-fos, c-myc and thymidylate synthase (TS) RNA constitutively, but at comparable levels to their parental counterparts. Whilst the ovarian carcinoma cells inherently expressed markedly higher levels (30- to 50-fold) of the excision repair gene ERCC1 than the teratoma cells, only the teratoma DXR10 subline showed an increased level of expression of ERCC1 mRNA relative to their parental cells. Expression of the ERCC3/XPB gene encoding a repair helicase, however, was similar in all the lines tested. The results suggest that CDDP resistance may be mediated by different mechanisms in these DXR10 sublines from those previously identified in drug-selected CDDP-resistant human ovarian A2780/DDP cells.
Subject(s)
Cisplatin/pharmacology , DNA Repair/genetics , Ovarian Neoplasms/drug therapy , Ovarian Neoplasms/genetics , Teratoma/drug therapy , Teratoma/genetics , Testicular Neoplasms/drug therapy , Testicular Neoplasms/genetics , DNA Damage , Drug Resistance/genetics , Female , Gene Expression/radiation effects , Genes, fos , Genes, myc , Humans , Male , Ovarian Neoplasms/enzymology , RNA, Neoplasm/biosynthesis , RNA, Neoplasm/metabolism , Teratoma/enzymology , Testicular Neoplasms/enzymology , Thymidylate Synthase/genetics , Tumor Cells, Cultured/radiation effectsABSTRACT
Some observations have been made on inferior olive specimens in the adult cat, with aldehydes fixation techniques through vascular perfusion suitable for electron microscropy. The results made it possible to recognise five types of vesicular structures present in different nerve endings (differences either in type or in number). A possible topographic organization of synaptic connections, between the afferent endings and the dendritic structures of the inferior olive has been considered and it seems that on every large dendrite, nearer the cell body, there is a development of numerous connection. As such connections give a great deal of information to the dendrites with much much interaction it seems possible that there is modulation in the activation of the postsynaptic element. On the other hand, in the periphery of the dendritic branch, many thin branches seem to make contact with few and large endings; this leads us to believe that the regulations of the information is not based only on the afferences but mainly on distribution, origin and number of the postsynaptic elements. A study of the interdendritic connections and of tubulo-vesicular structures showed that their diameter and dense content have a similar morphology with the big granulated vesicles, visible in the terminals, or along the fibres or, frequently, in the Golgi apparatus.
Subject(s)
Cats/anatomy & histology , Olivary Nucleus/ultrastructure , Animals , Microscopy, Electron , Nerve Endings/ultrastructure , Synapses/ultrastructureABSTRACT
NIH-3T3 fibroblasts were co-transfected with pSV2neo and pSG5tag by the calcium-phosphate precipitation method. The stable integration of the tag gene sequence and its transcription was verified by Southern and Northern blot analysis. 3-Methyladenine glycosylase activity in pSG5tag transfected 3T3 cells was approximately 400 times higher than in cells transfected with the control plasmid pSG5 or in the parental cells and was inhibited by 3-methyladenine. Bacterial tag gene can thus be expressed in mammalian cells and the encoded enzyme is functionally active. These transfected cells could serve as an important tool to investigate the importance of the repair of N3-adenine as a mechanism of protection against the mutagenicity and cytotoxicity of alkylating agents.
Subject(s)
DNA Glycosylases , Escherichia coli/genetics , N-Glycosyl Hydrolases/biosynthesis , Transfection , 3T3 Cells/metabolism , Animals , Cell Division , DNA Repair , Mice , N-Glycosyl Hydrolases/genetics , RNA, Messenger/analysis , Recombinant Proteins/biosynthesisABSTRACT
Treatment of C57Bl or BALB/C mice with human interferon-alpha A/D (HuIFN-alpha A/D) significantly increased hepatic levels of the DNA repair enzyme O6-alkylguanine DNA alkyltransferase (AT). The maximum induction was seen 24 h after a single dose of 50-100 micrograms/kg IFN-alpha A/D. No induction was observed in rat liver hepatocytes cultured in vitro. Liver AT was also induced by poly(I:C), which is a potent IFN inducer. By increasing AT levels, IFN could protect against the potentially mutagenic alkylation at guanine O6 position caused by some carcinogens. Moreover this finding suggests a link between immune response and the DNA repair system, possibly acting in concert to defend the body from potentially toxic compounds.