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Translational Therapeutics

Cytochrome P450 1B1 polymorphism drives cancer cell stemness and patient outcome in head-and-neck carcinoma




Cytochrome P450 1B1 (CYP1B1) is mostly expressed in tumours and displays unusual properties. Its two polymorphic forms were differently associated with anticancer drug sensitivity. We decipher here the role of this polymorphism in anticancer drug efficacy in vitro, in vivo and in the clinical setting.


From head-and-neck squamous cell carcinoma cell lines not expressing CYP1B1, we generated isogenic derivatives expressing the two forms. Proliferation, invasiveness, stem cell characteristics, sensitivity to anticancer agents and transcriptome were analysed. Tumour growth and chemosensitivity were studied in vivo. A prospective clinical trial on 121 patients with advanced head-and-neck cancers was conducted, and a validation-retrospective study was conducted.


Cell lines expressing the variant form displayed high rates of in vitro proliferation and invasiveness, stemness features and resistance to DNA-damaging agents. In vivo, tumours expressing the variant CYP1B1 had higher growth rates and were markedly drug-resistant. In the clinical study, overall survival was significantly associated with the genotypes, wild-type patients presenting a longer median survival (13.5 months) than the variant patients (6.3 months) (p = 0.0166).


This frequent CYP1B1 polymorphism is crucial for cancer cell proliferation, migration, resistance to chemotherapy and stemness properties, and strongly influences head-and-neck cancer patients’ survival.

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Fig. 1: Proliferation and migration capacity of the isogenic CYP1B1 cell lines.
Fig. 2: Cytotoxicity profiles of the isogenic cell lines.
Fig. 3: In vivo tumour growth of isogenic cell lines in immunodeficient mice.
Fig. 4: Drug-induced tumour growth inhibition of CAL27-derived isogenic cell lines in immunodeficient mice.
Fig. 5: Stemness characteristics of CAL27 CYP1B1-WT and CYP1B1-VAR cells.
Fig. 6: Survival curves of advanced head-and-neck cancer patients treated with chemotherapy plus cetuximab.


  1. 1.

    Laroche-Clary, A., Le Morvan, V., Yamori, T. & Robert, J. Cytochrome P450 1B1 gene polymorphisms as predictors of anticancer drug activity: studies with in vitro models. Mol. Cancer Ther. 9, 3315–3321 (2010).

    CAS  Article  Google Scholar 

  2. 2.

    Beuten., J., Gelfond, J. A., Byrne, J. J., Balic, I., Crandall, A. C., Johnson-Pais, T. L. et al. CYP1B1 variants are associated with prostate cancer in non-Hispanic and Hispanic Caucasians. Carcinogenesis 29, 1751–1757 (2008).

    CAS  Article  Google Scholar 

  3. 3.

    Chen, B., Qiu, L. X., Li, Y., Xu, W., Wang, X. L., Zhao, W. H. et al. The CYP1B1 Leu432Val polymorphism contributes to lung cancer risk: evidence from 6501 subjects. Lung Cancer 70, 247–252 (2010).

    Article  Google Scholar 

  4. 4.

    Salinas-Sanchez, A. S., Donate-Moreno, M. J., Lopez-Garrido, M. P., Gimenez-Bachs, J. M. & Escribano, J. Role of CYP1B1 gene polymorphisms in bladder cancer susceptibility. J. Urol. 187, 700–706 (2012).

    CAS  Article  Google Scholar 

  5. 5.

    Sissung, T. M., Danesi, R., Price, D. K., Steinberg, S. M., de Wit, R., Zahid, M. et al. Association of the CYP1B1*3 allele with survival in 8 patients with prostate cancer receiving docetaxel. Mol. Cancer Ther. 7, 19–26 (2008).

    CAS  Article  Google Scholar 

  6. 6.

    Le Morvan, V., Litière, S., Laroche-Clary, A., Ait-Ouferoukh, S., Bellott, R., Messina, C. et al. Identification of SNPs associated with response of breast cancer patients to neoadjuvant chemotherapy in the EORTC-10994 randomized phase III trial. Pharmacogenomics J. 15, 63–68 (2015).

    Article  Google Scholar 

  7. 7.

    Murray, G. I., Taylor, M. C., McFadyen, M. C., McKay, J. A., Greenlee, W. F., Burke, M. D. et al. Tumor-specific expression of cytochrome P450 CYP1B1. Cancer Res. 57, 3026–3031 (1997).

    CAS  PubMed  Google Scholar 

  8. 8.

    Murray, G. I., Melvin, W. T., Greenlee, W. F. & Burke, M. D. Regulation, function, and tissue-specific expression of cytochrome P450 CYP1B1. Annu. Rev. Pharmacol. Toxicol. 41, 297–316 (2001).

    CAS  Article  Google Scholar 

  9. 9.

    Gribben, J. G., Ryan, D. P., Boyajian, R., Urban, R. G., Hedley, M. L., Beach, K. et al. Unexpected association between induction of immunity to the universal tumor antigen CYP1B1 and response to next therapy. Clin. Cancer Res. 11, 4430–4436 (2005).

    CAS  Article  Google Scholar 

  10. 10.

    Cheever, M. A., Allison, J. P., Ferris, A. S., Finn, O. J., Hastings, B. M., Hecht, T. T. et al. The prioritization of cancer antigens: a National Cancer Institute pilot project for the acceleration of translational research. Clin. Cancer Res. 15, 5323–5337 (2009).

    Article  Google Scholar 

  11. 11.

    McFadyen, M. C., McLeod, H. L., Jackson, F. C., Melvin, W. T., Doehmer, J. & Murray, G. I. Cytochrome P450 CYP1B1 protein expression: a novel mechanism of anticancer drug resistance. Biochem. Pharmacol. 62, 207–212 (2001).

    CAS  Article  Google Scholar 

  12. 12.

    Huang, R. S., Duan, S., Kistner, E. O., Bleibel, W. K., Delaney, S. M., Fackenthal, D. L. et al. Genetic variants contributing to daunorubicin-induced cytotoxicity. Cancer Res. 68, 3161–3168 (2008).

    CAS  Article  Google Scholar 

  13. 13.

    Gioanni, J., Fischel, J. L., Lambert, J. C., Demard, F., Mazeau, C., Zanghellini, E. et al. Two new human tumor cell lines derived from squamous cell carcinomas of the tongue: establishment, characterization and response to cytotoxic treatment. Eur. J. Cancer Clin. Oncol. 24, 1445–1455 (1988).

    CAS  Article  Google Scholar 

  14. 14.

    Dull, T., Zufferey, R., Kelly, M., Mandel, R. J., Nguyen, M., Trono, D. et al. A third-generation lentivirus vector with a conditional packaging system. J. Virol. 72, 8463–8471 (1998).

    CAS  Article  Google Scholar 

  15. 15.

    Trapnell, C., Roberts, A., Goff, L., Pertea, G., Kim, D., Kelley, D. R. et al. Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks. Nat. Protoc. 7, 562–578 (2012).

    CAS  Article  Google Scholar 

  16. 16.

    Hu, Y. & Smyth, G. K. ELDA: extreme limiting dilution analysis for comparing depleted and enriched populations in stem cell and other assays. J. Immunol. Methods 347, 70–78 (2009).

    CAS  Article  Google Scholar 

  17. 17.

    Barretina, J., Caponigro, G., Stransky, N., Venkatesan, K., Margolin, A. A., Kim, S. et al. The cancer cell line encyclopedia enables predictive modelling of anticancer drug sensitivity. Nature 483, 603–607 (2012).

    CAS  Article  Google Scholar 

  18. 18.

    Kohn, K. W., Zeeberg, B. M., Reinhold, W. C. & Pommier, Y. Gene expression correlations in human cancer cell lines define molecular interaction networks for epithelial phenotype. PLoS ONE 9, e99269 (2014).

    Article  Google Scholar 

  19. 19.

    Reinhold, W. C., Varma, S., Sousa, F., Sunshine, M., Abaan, O. D., Davis, S. R. et al. NCI-60 whole exome sequencing and pharmacological CellMiner analyses. PLoS ONE 9, e101670 (2014).

    Article  Google Scholar 

  20. 20.

    Kalluri, R. & Weinberg, R. A. The basics of epithelial-mesenchymal transition. J. Clin. Invest. 119, 1420–1428 (2009).

    CAS  Article  Google Scholar 

  21. 21.

    Zhao, Y., Sorenson, C. M. & Sheibani, N. Cytochrome P450 1B1 and primary congenital glaucoma. J. Ophthalmic Vis. Res. 10, 60–67 (2015).

    Article  Google Scholar 

  22. 22.

    Yu, P. J., Chen, W. G., Feng, Q. L., Chen, W., Jiang, M. J. & Li, Z. Q. Association between CYP1B1 gene polymorphisms and risk factors and susceptibility to laryngeal cancer. Med. Sci. Monit. 21, 239–245 (2015).

    CAS  Article  Google Scholar 

  23. 23.

    De Iuliis, F., Salerno, G., Taglieri, L. & Scarpa, S. Are pharmacogenomic biomarkers an effective tool to predict taxane toxicity and outcome in breast cancer patients? Literature review. Cancer Chemother. Pharmacol. 76, 679–690 (2015).

    Article  Google Scholar 

  24. 24.

    Shimada, T., Watanabe, J., Inoue, K., Guengerich, F. P. & Gillam, E. M. Specificity of 17beta-oestradiol and benzo[a]pyrene oxidation by polymorphic human cytochrome P4501B1 variants substituted at residues 48, 119 and 432. Xenobiotica 31, 163–176 (2001).

    CAS  Article  Google Scholar 

  25. 25.

    Hanna, I. H., Dawling, S., Roodi, N., Guengerich, F. P. & Parl, F. F. Cytochrome P450 1B1 (CYP1B1) pharmacogenetics: association of polymorphisms with functional differences in estrogen hydroxylation activity. Cancer Res. 60, 3440–3444 (2000).

    CAS  PubMed  Google Scholar 

  26. 26.

    Aklillu, E., Øvrebø, S., Botnen, I. V., Otter, C. & Ingelman-Sundberg, M. Characterization of common CYP1B1 variants with different capacity for benzo[a]pyrene-7,8-dihydrodiol epoxide formation from benzo[a]pyrene. Cancer Res. 65, 5105–5111 (2005).

    CAS  Article  Google Scholar 

  27. 27.

    Wang, A., Savas, U., Stout, C. D. & Johnson, E. F. Structural characterization of the complex between alpha-naphthoflavone and human cytochrome P450 1B1. J. Biol. Chem. 286, 5736–5743 (2011).

    CAS  Article  Google Scholar 

  28. 28.

    Kwon, Y. J., Baek, H. S., Ye, D. J., Shin, S., Kim, D. & Chun, Y. J. CYP1B1 enhances cell proliferation and metastasis through induction of EMT and activation of Wnt/β-catenin signaling via Sp1 upregulation. PLoS ONE 11, e0151598 (2016).

    Article  Google Scholar 

  29. 29.

    Hunter, K. Host genetics influence tumour metastasis. Nat. Rev. Cancer 6, 141–146 (2006).

    CAS  Article  Google Scholar 

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We are grateful to Dr Laurence Bresson-Bepoldin and Dr Frédéric Delom for their kind support during the preparation of the paper. We are indebted to Prof Jean-Pierre Delord for his implication in the clinical validation study.

Author information




V.L.M. and J.R. designed the experiments and evaluated and interpreted the results. V.L.M. and A.La generated the isogenic cell lines. V.L.M. conducted most of the experiments and supervised those realised by younger scientists, M.C. and L.B.B. C.A. and D.F. realised part of the in vitro experiments. E.R. conducted all in vivo experiments. A.P. conducted the original prospective clinical trial that was analysed and validated by S.M.P. A.M. and A.Lu identified the patients and samples of the retrospective clinical validation study. V.L.M. and J.R. conceived and designed the project, planned the experiments, prepared the figures and wrote the paper with the help of D.F.

Corresponding author

Correspondence to Jacques Robert.

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Ethics approval and consent to participate

CAL27 and CAL33 HNSCC cell lines were kindly provided by Dr J.L. Fischel (Centre Antoine Lacassagne, Nice, France, where they were initially obtained). They were authenticated by Azur Génétique (Nice, France) as identical to the corresponding DSMZ cell lines (report reference AGLC-14-00225). The study was approved by the local ethical committee (Comité de Protection des Personnes de Bordeaux), and written informed consent was provided by all the participants after a full explanation of the study was given to them. This study was performed in accordance with the Declaration of Helsinki and was registered under ref. NCT-01827956.

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Not applicable.

Data availability

Raw results of the RNA-seq are available on the Annotare platform (ArrayExpress accession #E-MTAB-8512).

Competing interests

The authors declare no competing interests.

Funding information

This work was made possible through grants from the Ligue Nationale contre le Cancer (comité des Landes) and from GEFLUC-Aquitaine.

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Morvan, V.L., Richard, É., Cadars, M. et al. Cytochrome P450 1B1 polymorphism drives cancer cell stemness and patient outcome in head-and-neck carcinoma. Br J Cancer 123, 772–784 (2020).

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