Alter Cancer Susceptibility Research Articles

The biology of mammalian multi-copper ferroxidases.

The mammalian multicopper ferroxidases (MCFs) ceruloplasmin (CP), hephaestin (HEPH) and zyklopen (ZP) comprise a family of conserved enzymes that are essential for body iron homeostasis. Each of these enzymes contains six biosynthetically incorporated copper atoms which act as intermediate electron acceptors, and the oxidation of iron is associated with the four electron reduction of dioxygen to generate two water molecules. CP occurs in both a secreted and GPI-linked (membrane-bound) form, while HEPH and ZP each contain a single C-terminal transmembrane domain. These enzymes function to ensure the efficient oxidation of iron so that it can be effectively released from tissues via the iron export protein ferroportin and subsequently bound to the iron carrier protein transferrin in the blood. CP is particularly important in facilitating iron release from the liver and central nervous system, HEPH is the major MCF in the small intestine and is critical for dietary iron absorption, and ZP is important for normal hair development. CP and HEPH (and possibly ZP) function in multiple tissues. These proteins also play other (non-iron-related) physiological roles, but many of these are ill-defined. In addition to disrupting iron homeostasis, MCF dysfunction perturbs neurological and immune function, alters cancer susceptibility, and causes hair loss, but, despite their importance, how MCFs co-ordinately maintain body iron homeostasis and perform other functions remains incompletely understood.

Oral microbiome associated with lymph node metastasis in oral squamous cell carcinoma

Oral microbiota can alter cancer susceptibility and progression by modulating metabolism and inflammation. We assessed the association between the oral microbiome and lymph node (LN) metastasis in oral squamous cell carcinoma (OSCC). We collected a total of 54 saliva samples from patients with OSCC before surgery. LN metastasis was assessed based on postoperative pathological examination. We used QIIME2, linear discriminant analysis effect size (LEfSe), and PICRUSt2 methods to analyze microbial dysbiosis. A random forest classifier was used to assess whether the oral microbiome could predict LN metastasis. Among the 54 OSCC samples, 20 had LN metastasis, and 34 had no evidence of metastasis. There was a significant difference in β-diversity between the metastasis and no metastasis groups. Through LEfSe analysis, the metastasis group was enriched in the genera Prevotella, Stomatobaculum, Bifidobacterium, Peptostreptococcaceae, Shuttleworthia and Finegoldia. Pathways related to signal peptidase II were predominant in the no metastasis group. The RF model showed a modestly high accuracy for predicting metastasis. Differences in microbial community composition and functions were observed in the oral microbiome of patients with OSCC with and without LN metastasis. However, the finding that specific taxa may be associated with LN metastasis should be verified in a further prospective study.

Tissue-specific microRNA expression alters cancer susceptibility conferred by a TP53 noncoding variant

A noncoding polymorphism (rs78378222) in TP53, carried by scores of millions of people, was previously associated with moderate risk of brain tumors and other neoplasms. We find a positive association between this variant and soft tissue sarcoma. In sharp contrast, it is protective against breast cancer. We generated a mouse line carrying this variant and found that it accelerates spontaneous tumorigenesis and glioma development, but strikingly, delays mammary tumorigenesis. The variant creates a miR-382-5p targeting site and compromises a miR-325-3p site. Their differential expression results in p53 downregulation in the brain, but p53 upregulation in the mammary gland of polymorphic mice compared to that of wild-type littermates. Thus, this variant is at odds with Li-Fraumeni Syndrome mutants in breast cancer predisposition yet consistent in glioma predisposition. Our findings elucidate an underlying mechanism of cancer susceptibility that is conferred by genetic variation and yet altered by microRNA expression.

Genetic Ablation of Rbm38 Promotes Lymphomagenesis in the Context of Mutant p53 by Downregulating PTEN.

Mutant p53 exerts gain-of-function effects that drive metastatic progression and therapeutic resistance, but the basis for these effects remain obscure. The RNA binding protein RBM38 limits translation of mutant p53 and is often altered in tumors harboring it. Here we show how loss of Rbm38 significantly alters cancer susceptibility in mutant p53 knock-in mice by shortening lifespan, altering tumor incidence, and promoting T-cell lymphomagenesis. Loss of Rbm38 enhanced mutant p53 expression and decreased expression of the tumor suppressor Pten, a key regulator of T-cell development. Furthermore, Rbm38 was required for Pten expression via stabilization of Pten mRNA through an AU-rich element in its 3'UTR. Our results suggest that Rbm38 controls T-cell lymphomagenesis by jointly modulating mutant p53 and Pten, with possible therapeutic implications for treating T-cell malignancies.Significance: An RNA-binding protein controls T-cell lymphomagenesis by jointly modulating mutant p53 and PTEN, with possible therapeutic implications for treating T-cell malignancies. Cancer Res; 78(6); 1511-21. ©2018 AACR.

The role of the microbiome in cancer development and therapy.

Answer questions and earn CME/CNE The human body harbors enormous numbers of microbiota that influence cancer susceptibility, in part through their prodigious metabolic capacity and their profound influence on immune cell function. Microbial pathogens drive tumorigenesis in 15% to 20% of cancer cases. Even larger numbers of malignancies are associated with an altered composition of commensal microbiota (dysbiosis) based on microbiome studies using metagenomic sequencing. Although association studies cannot distinguish whether changes in microbiota are causes or effects of cancer, a causative role is supported by rigorously controlled preclinical studies using gnotobiotic mouse models colonized with one or more specific bacteria. These studies demonstrate that microbiota can alter cancer susceptibility and progression by diverse mechanisms, such as modulating inflammation, inducing DNA damage, and producing metabolites involved in oncogenesis or tumor suppression. Evidence is emerging that microbiota can be manipulated for improving cancer treatment. By incorporating probiotics as adjuvants for checkpoint immunotherapy or by designing small molecules that target microbial enzymes, microbiota can be harnessed to improve cancer care. CA Cancer J Clin 2017;67:326-344. © 2017 American Cancer Society.

A systematic review and meta-analysis of the association between OGG1 Ser326Cys polymorphism and cancers.

The oxyguanine glycosylase 1 (OGG1) gene has an important role in DNA repair, and the polymorphism of the gene may alter cancer susceptibility. This study aims to examine the association between the OGG1 Ser326Cys polymorphism and cancer risk based on meta-analysis. Relevant studies were identified through a search of PubMed and Weipu databases, and a total of 109 studies including 111 comparisons containing 34,041 cases and 42,730 controls were enrolled. Overall, significant association was observed between OGG1 Ser326Cys polymorphism and cancer risk in all genetic models except for heterozygote model (Cys/Cys+Cys/Ser vs Ser/Ser: OR 1.071, 95% CI 1.019-1.125; Cys/Cys vs Cys/Ser+Ser/Ser: OR 1.159, 95% CI 1.076-1.248; Cys/Cys vs Ser/Ser: OR 1.202, 95% CI 1.105-1.308). In stratified analysis by cancer type, significantly increased cancer risk was observed in digestive system cancer, head and neck cancer and lung cancer. For gynecologic cancer, significantly increased cancer risk was also observed in homozygote model (OR 1.974, 95% CI 1.254-3.107). In addition, in stratified analysis by ethnicities, increased cancer risk was found in Asians (Cys/Cys vs Cys/Ser+Ser/Ser: OR 1.195, 95% CI 1.088-1.313; Cys/Cys+Cys/Ser vs Ser/Ser: OR 1.115, 95% CI 1.045-1.190; Cys/Cys vs Ser/Ser: OR 1.273, 95% CI 1.149-1.410). The OGG1 Ser326Cys polymorphism may be a risk factor for cancers of lung, digestive system and head and neck.

Abstract C61: Genetic alterations in the TP53 genomic region of African American and Caucasian colorectal cancers

Abstract Purpose: In the US, colorectal cancer (CRC) is the second leading cause of cancer-related deaths and the third most common cancer in both men and women. CRC affects African Americans (AAs) disproportionately, relative to non-Hispanic Caucasians (CAs). Data from our prior studies indicate that the p53 codon 72 polymorphism is disproportionately higher in AAs compared to CAs [Clin Cancer Res; 15(14); 2406–-2416. 2009]. It is also evident that genetic human diversity is due not only to single nucleotide polymorphisms (SNPs), but also to structural variants or retrotransposons. Retrotransposons induce mutations, near or within genes by several mechanisms: insertions, deletions, duplications, copy number variants, inversions, or translocations, all of which expand human diversity and possibly alter cancer susceptibility. Thus, our objectives are to establish genomic profiles of the p53 tumor suppressor (TP53) region of CRCs of AA and CA patients to identify race/ethnic-specific alterations that are associated with ancestry, and to develop a DNA sequencing protocol to delineate AA and CA patient profiles using this complex genomic region (∼19 Kb). Methods: Genomic DNA was extracted from 4 frozen tissue samples (2 CRCs and their corresponding normal pairs) and 4 (2 CRCs/normal pairs) formalin-fixed, paraffin-embedded (FFPE) tissues. For the DNA of two CRC patients (one tumor/normal pair each of AA and CA patients), the complete TP53 gene (∼19 Kb, including exons and introns) was amplified by PCR in two halves, followed by Illumina next-generation sequencing and comparison to the human genome 19 reference sequence. For each patient, paired normal (benign/control) and tumor tissue DNAs were compared. Results: TheTP53 gene was amplified from DNA extracted from all 4 frozen CRC tissues, but not from the 4 FFPE tissues. Sequence comparisons of normal versus tumor DNA revealed 80 single nucleotide polymorphisms (SNPs). Most (96%, n=77) located within introns. Of these, 34 were shared by AA and CA patients. In contrast, 8 SNPs were detected only in a CA patient, and 13 others were present only in an AA patient. Two SNPs found in our AA patient were absent in our CA patient and in the publicly available HapMap database of CAs, suggesting that these SNPs reflect African ancestry. Also, 22 SNPs were exclusively present in the CA tumor, whereas only 3 SNPs were unique to the AA tumor. These findings deserve further investigation. Conclusions: We developed PCR protocols that should allow us to conduct a comprehensive mutational profiling of coding and non-coding regions of the TP53 genomic region in DNA isolated from frozen tissues. Comparison of genetic alteration profiles of TP53 in AAs and CAs will aid in determining the race/ethnicity of CRC patients. These studies were supported by a Charles Barkley Foundation grant though the UAB MHRC and a pre-pilot project of the UAB/TU/MSM Partnership grant, U54-CA 118948. Citation Format: Jesus F. Salazar-Gonzalez, Maria G. Salazar, Balananda Dhurjati Kumar Putcha, Edward E. Partridge, Mona N. Fouad, Manne Upender. Genetic alterations in the TP53 genomic region of African American and Caucasian colorectal cancers. [abstract]. In: Proceedings of the Sixth AACR Conference: The Science of Cancer Health Disparities; Dec 6–9, 2013; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2014;23(11 Suppl):Abstract nr C61. doi:10.1158/1538-7755.DISP13-C61

An updated meta-analysis on association between angiotensin I-converting enzyme gene insertion/deletion polymorphism and cancer risk

The Alu repetitive sequence insertion/deletion (I/D, rs4646994) polymorphism in the angiotensin I-converting enzyme (ACE) gene may alter cancer susceptibility, but results of current studies are inconclusive. To derive a more precise estimation of the relationship between the ACE I/D polymorphism and cancer risk, we performed an updated meta-analysis of all eligible studies. All studies published up to July 2013 concerning the association between the ACE I/D polymorphism and cancer risk were identified by systematically searching PubMed, EMBASE, Wanfang, CNKI, and Cqvip databases. The odds ratios (ORs) with 95 % confidence intervals (CIs) were pooled using the fixed/random-effects model in Review Manager 5.1 and STATA 12.0. A total of 46 case-control studies including 7,025 cases and 34,911 controls were identified and evaluated. Overall, we did not observe a direct association between the ACE I/D polymorphism and general cancer risk (DD + DI vs. II OR = 0.95, 95 %CI = 0.84-1.07, P = 0.40). In the subgroup analysis by cancer type, a significant increased susceptibility of prostate cancer was found for variant homozygotes (DD vs. II + ID OR = 2.15, 95 %CI = 1.01-4.55, P = 0.05). Additionally, no significant association was observed in other subgroup analyses according to ethnicity, control source, sample size and quality control of genotyping. In summary, our results suggested that the ACE I/D polymorphism might not be a common risk factor for overall cancer susceptibility, but might contribute to the susceptibility of prostate cancer. More studies with larger sample sizes are required in the future.

Abstract 881: Polymorphisms of PUMA, a BH3-only pro-apoptotic gene, modify risk of squamous cell carcinoma of head and neck associated with HPV16 infection

Abstract PUMA (p53 upregulated modulator of apoptosis) and Noxa (Latin for damage), as well as their common target MCL-1 (myeloid cell leukemia 1), respond to human papillomavirus 16 (HPV16) oncoprotein E6-mediated p53 degradation or E7-mediated pRb inactivation, and therefore may alter cancer susceptibility through the PUMA (Noxa)-MCL1-apoptosis mechanism. In this case-control study of 380 cases and 335 cancer-free matched controls, we evaluated HPV16 seropositivity and two single nucleotide polymorphisms (SNPs) in the promoter region for each gene (rs3810294 and rs2032809 for PUMA, rs4558496 and rs9957673 for Noxa, rs9803935 and rs3738485 for MCL1) to examine their possible associations with risk of squamous cell carcinoma of head and neck (SCCHN). Of the 6 SNPs, both two SNPs of PUMA were significantly associated with 1.5-fold increased SCCHN risk in a dominant model. When we include the HPV16 seropositivity in the model, the risk of SCCHN was significantly increased in an allele-dose effect manner (Ptrend< 0.0001 for each SNP, respectively). Notably, in the site-stratified analysis, oropharyngeal cancer risk was dramatically increased, particularly among minor alleles carriers having the HPV16 L1 seropositivity with OR (95%CI) of 13.8 (5.4-35.5) for rs3810294 and 8.1 (4.3-15.0) for rs2032809 (both Ptrend < 0.0001). These data suggest some synergistic effects between PUMA variants and HPV16 seropositivity on susceptibility to SCCHN, especially in those cases with HPV-related oropharyngeal cancer. Additionally, the modifying effect of PUMA variants on oropharyngeal cancer associated with the HPV16 L1 seropositivity was more predominant in never-smokers, never-drinkers and young subjects. Our results suggest that PUMA promoter polymorphisms, synergistically interacting with HPV16 L1 seropositivity, modify risk of HPV-related oropharyngeal cancer. Further molecular and epidemiologic studies with larger samples are needed to validate our findings. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 881. doi:10.1158/1538-7445.AM2011-881

Interactions between cigarette smoking and selected polymorphisms in xenobiotic metabolizing enzymes in risk for colorectal cancer: A case‐only analysis

The metabolism of xenobiotics is complex and involves multiple steps and multiple enzymes. Genetic variation in the genes encoding these enzymes as well as the level of exposure to the substrates of these enzymes could alter metabolism and clearance of potential carcinogens and thus alter cancer susceptibility. This study examined interaction effect between smoking and two single nucleotide polymorphisms (SNPs)-CYP1A1 c.1384A>G (p.Ile462Val) and EPHX1 c.337T>C (p.Tyr113His)-in modulating colorectal cancer (CRC) risk. The SNPs were selected a priori based on functional significance. In a case-only analysis, unconditional logistic regression was used to examine the associations between smoking and each SNP and between the two SNPs in 786 patients with nonfamilial CRC. There was significant multiplicative interaction for CRC risk between smoking and EPHX1 c.337T>C (odds ratio [OR] = 1.37, 95% confidence interval [CI] = 1.03-1.81, P = 0.03), particularly among smokers with a history of greater than 20 pack-years of smoking (OR = 1.52, 95% CI = 1.07-2.16, P = 0.02). In addition, there was gene-gene interaction between EPHX1 c.337T>C and CYP1A1 c.1384A>G (OR = 1.61, 95% CI = 1.02-2.55, P = 0.04). Smokers with any variant allele of EPHX1 were at increased risk for CRC, as were individuals with any variant allele of CYP1A1 together with any variant allele of EPHX1. Thus, the study of gene-environment and gene-gene interactions may help to identify high-risk subgroups that can be targeted for intensive smoking cessation and CRC screening interventions.

Genomic DNA Methylation among Women in a Multiethnic New York City Birth Cohort

One plausible mechanism for the environment to alter cancer susceptibility is through DNA methylation. Alterations in DNA methylation can lead to genomic instability and altered gene transcription. Genomic DNA methylation levels have been inversely associated with age, suggesting that factors throughout life may be associated with declines in DNA methylation. Using information from a multiethnic New York City birth cohort (born between 1959 and 1963), we examined whether genomic DNA methylation, measured in peripheral blood mononuclear cells, was associated with smoking exposure and other epidemiologic risk factors across the life course. Information on prenatal and childhood exposures was collected prospectively through 1971, and information on adult exposures and blood specimens were collected in adulthood from 2001 to 2007. Methylation levels of leukocyte DNA were determined using a [(3)H]-methyl acceptance assay where higher values of disintegrations per minute per microgram DNA indicate less DNA methylation. Genomic methylation of leukocyte DNA differed by ethnicity (66% of Blacks, 48% of Whites, and 29% of Hispanics were above the median level of disintegrations per minute per microgram DNA; P = 0.03). In multivariable modeling, DNA methylation was statistically significantly associated with maternal smoking during pregnancy, longer birth length, later age at menarche, nulliparity, and later age at first birth. These data, if replicated in larger samples, suggest that risk factors across the life course may be associated with DNA methylation in adulthood. Larger studies and studies that measure within-individual changes in DNA methylation over time are a necessary next step.

Common genetic variants in pre-microRNAs were associated with increased risk of breast cancer in Chinese women

Small, noncoding RNA molecules, called microRNAs (miRNAs), are thought to function as either tumor suppressors or oncogenes. Common single-nucleotide polymorphisms (SNPs) in miRNAs may change their property through altering miRNA expression and/or maturation, and thus they may have an effect on thousands of target mRNAs, resulting in diverse functional consequences. However, it remains largely unknown whether miRNA SNPs may alter cancer susceptibility. We evaluated the associations of selected four SNPs (rs2910164, rs2292832, rs11614913, and rs3746444) in pre-miRNAs (hsa-mir-146a, hsa-mir-149, hsa-mir-196a2, and hsa-mir-499) with breast cancer risk in a case-control study of 1,009 breast cancer cases and 1,093 cancer-free controls in a population of Chinese women and we found that hsa-mir-196a2 rs11614913:T>C and hsa-mir-499 rs3746444:A>G variant genotypes were associated with significantly increased risks of breast cancer (odds ratio [OR], 1.23; 95% confidence interval [CI], 1.02-1.48 for rs11614913:T>C; and OR, 1.25; 95% CI, 1.02-1.51 for rs3746444:A>G in a dominant genetic model) in a dose-effect manner (P for trend was 0.010 and 0.037, respectively). These findings suggest, for the first time, that common SNPs in miRNAs may contribute to breast cancer susceptibility. Further functional characterization of miRNA SNPs and their influences on target mRNAs may provide underlying mechanisms for the observed associations and disease etiology.

Lung cancer risk and variation in MGMT activity and sequence

O 6-Alkylguanine–DNA alkyltransferase (MGMT) repairs DNA adducts that result from alkylation at the O 6 position of guanine. These lesions are mutagenic and toxic and can be produced by a variety of agents including the tobacco-specific nitrosamines, carcinogens present in cigarette smoke. Here, we review some of our work in the context of inter-individual differences in MGMT expression and their potential influence on lung cancer risk. In humans there are marked inter-individual differences in not only levels of DNA damage in the lung (N7-methylguanine) that can arise from exposure to methylating agents but also in MGMT activity in lung tissues. In the presence of such exposure, this variability in MGMT activity may alter cancer susceptibility, particularly as animal models have demonstrated that the complete absence of MGMT activity predisposes to alkylating-agent induced cancer while overexpression is protective. Recent studies have uncovered a series of polymorphisms that affect protein activity or are associated with differences in expression levels. The associations between these (and other) polymorphisms and cancer risk are inconsistent, possibly because of small sample sizes and inter-study differences in lung cancer histology. We have recently analysed a consecutive series of case–control studies and found evidence that lung cancer risk was lower in subjects with the R178 allele.

Rapid analysis of XRCC1 polymorphisms using real-time polymerase chain reaction

DNA repair plays a critical role in protecting the genome from carcinogens or ionizing radiation. Three coding polymorphisms at codons 194, 280, and 399 in X-ray cross-complementing group 1 (XRCC1) DNA repair gene have been identified that may affect DNA repair and alter cancer susceptibility. In order to study their role in molecular-epidemiology studies we developed a single-step procedure for genotyping these polymorphisms using real-time polymerase chain reaction (rt-PCR) and subsequent melting curve analysis. Genotypes of 622 unrelated Caucasians without prior history of cancer were determined by real-time PCR and compared to genotypes obtained by restriction fragment length polymorphism PCR. In the population studied, the allele frequency of the XRCC1 26304 site (C-->T) of codon 194 in exon 6 was 0.065, the allele frequency of the XRCC1 27466 site (G-->A) of codon 280 in exon 9 was 0.048 and of the XRCC1 28152 site (G-->A) of codon 399 in exon 10 was 0.35. There was no disagreement between the two methods. These findings confirm the real-time fluorescence PCR method as a rapid and reliable assay for the analysis of large numbers of samples.

XPA, haplotypes, and risk of basal and squamous cell carcinoma

Nucleotide excision repair (NER) is instrumental in removing DNA lesions caused by ultraviolet (UV) radiation, the dominant risk factor for keratinocyte carcinoma, including basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). We evaluated whether BCC or SCC risk was influenced by the A23G single nucleotide polymorphism (SNP) in Xeroderma pigmentosum group A (XPA), which codes for an essential protein in NER. We also investigated whether haplotypes of XPA, determined by seven haplotype-tagging SNPs, better define susceptibility to keratinocyte carcinoma. Incident cases of BCC and SCC from New Hampshire were identified through dermatologists and pathology laboratories. Population-based controls were frequency-matched to cases by gender and age. Cases of BCC (886) and of SCC (682) were compared with controls (796). Models controlled for age, gender, pigmentation factors and severe sunburns and were restricted to Caucasians. Using GG as the reference, the A allele was less frequent among cases of BCC (OR(AG) = 0.82, 95% CI (0.66, 1.01); OR(AA)= 0.74, 95% CI (0.53, 1.03); trend test P = 0.03) and SCC (OR(AG) = 0.85, 95% CI (0.67, 1.07); OR(AA) = 0.74, 95% CI (0.52, 1.05); trend test P = 0.05) than controls. Risk from > or =3 severe sunburns was elevated for those with the GG genotype only, and this interaction was nearly significant for BCC (P = 0.07). XPA genotype also modified a relationship between SCC and the amount of pigmentation (P = 0.02). Using a haplotype analysis identifying seven common XPA haplotypes indicated that the A23G polymorphism alone captured the differences in susceptibility to keratinocyte carcinoma. The common G allele of the A23G polymorphism was associated with an increased risk of BCC and SCC and this polymorphism appeared to be the determining polymorphism in XPA that alters cancer susceptibility.

Polymorphism of the Pi class glutathione S-transferase in normal populations and cancer patients.

Deficiencies of the glutathione transferase isoenzymes GSTM1-1 and GSTT1-1 have been shown to be risk modifiers in a number of different cancers but there have been no similar studies with GSTP1-1, the only member of the Pi class of glutathione S-transferases expressed in humans. Over-expression of GSTP1-1 in tumours suggests that it may be a significant factor in acquired resistance to certain anticancer drugs. We previously identified a cDNA clone with two amino acid substitutions (I105V, A114V). This clone suggests that the GSTP1 gene is polymorphic and it is possible that the different genotypes may be associated with altered cancer risk or drug resistance. In the present study, we report methods for genotyping individuals at codons 105 and 114 of GSTP1 and demonstrate that these two loci are polymorphic in several different racial groups. We also detected significant linkage disequilibrium between these two loci. To determine if either of the alleles at these two loci were associated with altered cancer susceptibility, we genotyped individuals with colorectal cancer or lung cancer. A total of 131 colorectal and 184 lung cancer patients were compared with 199 control individuals. Overall, there were no significant associations between the GSTP1 polymorphisms and either form of cancer.

Molecular genetics of the human cytochrome P450-dependent monooxygenases.

In this work, the role of genetic as well as environmental factors in determining cytochrome P450 isozyme levels in man have been studied. Simple DNA based assays for the identification of individuals nulled at the CYP2D6 locus are described and have been applied to investigate whether this gene defect is associated with altered cancer susceptibility. In contrast to literature reports, in no cancer type were poor metabolizers underrepresented, indeed in several cancers the mutant allele frequency was increased. A model using human tumours grown as xenografts is described that should help elucidate the factors which regulate P450 levels in man.