Metabolism of anticancer drugs affects their antitumor effects. This study has investigated the associations of gene expression of enzymes metabolizing anticancer drugs with therapy response and survival of breast carcinoma patients. Gene expression of 13 aldo-keto reductases (AKRs), carbonyl reductase 1, and 10 cytochromes P450 (CYPs) was assessed using quantitative real-time polymerase chain reaction in tumors and paired adjacent nonneoplastic tissues from 68 posttreatment breast carcinoma patients. Eleven candidate genes were then evaluated in an independent series of 50 pretreatment patients. Protein expression of the most significant genes was confirmed by immunoblotting. AKR1A1 was significantly overexpressed and AKR1C1-4, KCNAB1, CYP2C19, CYP3A4, and CYP3A5 downregulated in tumors compared with control nonneoplastic tissues after correction for multiple testing. Significant association of CYP2B6 transcript levels in tumors with expression of hormonal receptors was found in the posttreatment set and replicated in the pretreatment set of patients. Significantly higher intratumoral levels of AKR1C1, AKR1C2, or CYP2W1 were found in responders to neoadjuvant chemotherapy compared with nonresponders. Patients with high AKR7A3 or CYP2B6 levels in the pretreatment set had significantly longer disease-free survival than patients with low levels. Protein products of AKR1C1, AKR1C2, AKR7A3, CYP3A4, and carbonyl reductase (CBR1) were found in tumors and those of AKR1C1, AKR7A3, and CBR1 correlated with their transcript levels. Small interfering RNA-directed knockdown of AKR1C2 or vector-mediated upregulation of CYP3A4 in MDA-MB-231 model cell line had no effect on cell proliferation after paclitaxel treatment in vitro. Prognostic and predictive roles of drug-metabolizing enzymes strikingly differ between posttreatment and pretreatment breast carcinoma patients. Mechanisms of action of AKR1C2, AKR7A3, CYP2B6, CYP3A4, and CBR1 should continue to be further followed in breast carcinoma patients and models.

From the Toxicogenomics Unit VELAB Ltd Zlin Czech Republic

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Ferlay J, Shin HR, Bray F, et al. GLOBOCAN 2008, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 10. Lyon, France: International Agency for Research on Cancer; 2010.

Baguley BC. Multiple drug resistance mechanisms in cancer. Mol Biotechnol 2010; 46:308–316. PubMed

Johansson I, Ingelman-Sundberg M. Genetic polymorphism and toxicology with emphasis on cytochrome P450. Toxicol Sci 2011; 120:1–13. PubMed

Roy P, Yu LJ, Crespi CL, et al. Development of a substrate-activity based approach to identify the major human liver P-450 catalysts of cyclophosphamide and ifosfamide activation based on cDNA-expressed activities and liver microsomal P-450 profiles. Drug Metab Dispos 1999; 27:655–666. PubMed

Chang TK, Weber GF, Crespi CL, et al. Differential activation of cyclophosphamide and ifosphamide by cytochromes P-450 2B and 3A in human liver microsomes. Cancer Res 1993; 53:5629–5637. PubMed

Rodriguez-Antona C, Gomez A, Karlgren M, et al. Molecular genetics and epigenetics of the cytochrome P450 gene family and its relevance for cancer risk and treatment. Hum Genet 2010; 127:1–17. PubMed

Rahman A, Korzekwa KR, Grogan J, et al. Selective biotransformation of taxol to 6α-hydroxytaxol by human cytochrome P450 2C8. Cancer Res 1994; 54:5543–5546. PubMed

Shou M, Korzekwa KR, Krausz KW, et al. Role of human cytochrome P450 3A4 and 3A5 in the metabolism of Taxotere and its derivatives: enzyme specificity, interindividual distribution and metabolic contribution in human liver. Pharmacogenetics 1998; 8:391–401. PubMed

Komatsu T, Yamazaki H, Shimada N, et al. Roles of cytochromes P450 1A2, 2A6, and 2C8 in 5-fluorouracil formation from tegafur, an anticancer prodrug, in human liver microsomes. Drug Metab Dispos 2000; 28:1457–1463. PubMed

Sim SC, Kacevska M, Ingelman-Sundberg M. Pharmacogenomics of drug-metabolizing enzymes: a recent update on clinical implications and endogenous effects. Pharmacogenomics J 2013; 13:1–11. PubMed

Iscan M, Klaavuniemi T, Coban T, et al. The expression of cytochrome P450 enzymes in human breast tumours and normal breast tissue. Breast Cancer Res Treat 2001; 70:47–54. PubMed

Modugno F, Knoll C, Kanbour-Shakir A, et al. A potential role for the estrogen-metabolizing cytochrome P450 enzymes in human breast carcinogenesis. Breast Cancer Res Treat 2003; 82:191–197. PubMed

Murray GI, Patimalla S, Stewar KN, et al. Profiling the expression of cytochrome P450 in breast cancer. Histopathology 2010; 57:202–211. PubMed

Karlgren M, Gomez A, Stark K, et al. Tumor-specific expression of the novel cytochrome P450 enzyme, CYP2W1. Biochem Biophys Res Commun 2006; 341:451–458. PubMed

Edler D, Stenstedt K, Ohrling K, et al. The expression of the novel CYP2W1 enzyme is an independent prognostic factor in colorectal cancer-a pilot study. Eur J Cancer 2009; 45:705–712. PubMed

Barski OA, Tipparaju SM, Bhatnagar A. The aldo-keto reductase superfamily and its role in drug metabolism and detoxification. Drug Metab Rev 2008; 40:553–624. PubMed PMC

Wermuth B, Platts KL, Seidel A, et al. Carbonyl reductase provides the enzymatic basis of quinone detoxication in man. Biochem Pharmacol 1986; 35:1277–1282. PubMed

Thorn CF, Oshiro C, Marsh S, et al. Adriamycin pathways: pharmacodynamics and adverse effects. Pharmacogenet Genomics 2011; 21:440–446. PubMed PMC

Hlaváč V, Brynychová V, Václavíková R, et al. The expression profile of ABC transporter genes in breast carcinoma. Pharmacogenomics 2013; 14:515–529. PubMed

Hubackova M, Vaclavikova R, Ehrlichova M, et al. Association of superoxide dismutases and NAD(P)H oxidoreductases with prognosis of patients with breast carcinomas. Int J Cancer 2012; 130:338–348. PubMed

Therasse P, Arbuck SG, Eisenhauer EA, et al. New guidelines to evaluate the response to treatment in solid tumors. European Organization for Research and Treatment of Cancer, National Cancer Institute of the United States, National Cancer Institute of Canada. J Natl Cancer Inst 2000; 92:205–216. PubMed

Bustin SA, Benes V, Garson JA, et al. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 2009; 55:611–622. PubMed

Anzenbacher P, Souček P, Gut I, et al. Presence and activity of cytochrome P-450 isoforms in minipig liver microsomes: comparison with human liver samples. Drug Metab. Dispos 1998; 26:56–59. PubMed

Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J Royal Statist Soc B 1995; 57:289–300.

Bièche I, Girault I, Urbain E, et al. Relationship between intratumoral expression of genes coding for xenobiotic-metabolizing enzymes and benefit from adjuvant tamoxifen in estrogen receptor alpha-positive postmenopausal breast carcinoma. Breast Cancer Res 2004; 6:R252–R263. PubMed PMC

Tozlu S, Girault I, Vacher S, et al. Identification of novel genes that co-cluster with estrogen receptor alpha in breast tumor biopsy specimens, using a large-scale real-time reverse transcription-PCR approach. Endocr Relat Cancer 2006; 13:1109–1120. PubMed

Lo R, Burgoon L, Macpherson L, et al. Estrogen receptor-dependent regulation of CYP2B6 in human breast cancer cells. Biochim Biophys Acta 2010; 1799:469–479. PubMed PMC

Schmidt R, Baumann F, Knüpfer H, et al. CYP3A4, CYP2C9 and CYP2B6 expression and ifosfamide turnover in breast cancer tissue microsomes. Br J Cancer 2004; 90:911–916. PubMed PMC

Reyaul K, Thumar JK, Lundgren DH, et al. Differential protein expression profiles in estrogen receptor–positive and –negative breast cancer tissues using label-free quantitative proteomics. Genes Cancer 2010; 1:251–271. PubMed PMC

Dunnwald LK, Rossing MA, Li CI. Hormone receptor status, tumor characteristics, and prognosis: a prospective cohort of breast cancer patients. Breast Cancer Res 2007; 9:R6. PubMed PMC

Lewis MJ, Wiebe JP, Heathcote JG. Expression of progesterone metabolizing enzyme genes (AKR1C1, AKR1C2, AKR1C3, SRD5A1, SRD5A2) is altered in human breast carcinoma. BMC Cancer 2004; 4:27. PubMed PMC

Ji Q, Aoyama C, Nien YD, et al. Selective loss of AKR1C1 and AKR1C2 in breast cancer and their potential effect on progesterone signaling. Cancer Res 2004; 64:7610–7617. PubMed

Veitch ZW, Guo B, Hembruff SL, et al. Induction of 1C aldoketoreductases and other drug dose-dependent genes upon acquisition of anthracycline resistance. Pharmacogenet Genomics 2009; 19:477–488. PubMed

Wu L, Hu X, Tang H, et al. Valid application of Western blotting. Mol Biol Rep 2014; 41:3517–3520. PubMed

Miyoshi Y, Ando A, Takamura Y, et al. Prediction of response to docetaxel by CYP3A4 mRNA expression in breast cancer tissues. Int J Cancer 2002; 97:129–132. PubMed

Miyoshi Y, Taguchi T, Kim SJ, et al. Prediction of response to docetaxel by immunohistochemical analysis of CYP3A4 expression in human breast cancers. Breast Cancer 2005; 12:11–15. PubMed

Kapucuoglu N, Coban T, Raunio H, et al. Immunohistochemical demonstration of the expression of CYP2E1 in human breast tumour and non-tumour tissues. Cancer Lett 2003; 196:153–159. PubMed

Kim SM, Acharya P, Engel JC, et al. Liver cytochrome P450 3A ubiquitination in vivo by gp78/autocrine motility factor receptor and C terminus of Hsp70-interacting protein (CHIP) E3 ubiquitin ligases: physiological and pharmacological relevance. J Biol Chem 2010; 285:35866–35877. PubMed PMC

Olson RD, Mushlin PS, Brenner DE, et al. Adriamycin cardiotoxicity may be caused by its metabolite, adriamycinol. Proc Natl Acad Sci USA 1988; 85:3585–3589. PubMed PMC

Lal S, Sandanaraj E, Wong ZW, et al. CBR1 and CBR3 pharmacogenetics and their influence on adriamycin disposition in Asian breast cancer patients. Cancer Sci 2008; 99:2045–2054. PubMed PMC

López de Cerain A, Marín A, Idoate MA, et al. Carbonyl reductase and NADPH cytochrome P450 reductase activities in human tumoral versus normal tissues. Eur J Cancer 1999; 35:320–324. PubMed

Murakami A, Yakabe K, Yoshidomi K, et al. Decreased carbonyl reductase 1 expression promotes malignant behaviours by induction of epithelial mesenchymal transition and its clinical significance. Cancer Lett 2012; 323:69–76. PubMed

Murakami A, Fukushima C, Yoshidomi K, et al. Suppression of carbonyl reductase expression enhances malignant behaviour in uterine cervical squamous cell carcinoma: carbonyl reductase predicts prognosis and lymph node metastasis. Cancer Lett 2011; 311:77–84. PubMed

Use of Germline Genetic Variability for Prediction of Chemoresistance and Prognosis of Breast Cancer Patients

Molecular profile of 5-fluorouracil pathway genes in colorectal carcinoma