Top

Published in:

01-05-2013 | Original Article

Minor role of pregnane-x-receptor for acquired multidrug resistance in head and neck squamous cell carcinoma in vitro

Authors: Juan Pablo Rigalli, Tasmin Reuter, Christel Herold-Mende, Gerhard Dyckhoff, Walter Emil Haefeli, Johanna Weiss, Dirk Theile

Published in: Cancer Chemotherapy and Pharmacology | Issue 5/2013

Abstract

Purpose

Acquired multidrug resistance (MDR) has been linked to overexpression of drug-metabolising and transporting proteins mediated by pregnane-x-receptor (PXR). The aim of this work was to establish the relevance of PXR for MDR in head and neck squamous cell carcinoma (HNSCC).

Methods

Using eight HNSCC cell lines, we determined the efficacy of paclitaxel, cisplatin and 5-fluorouracil (5-FU) via proliferation assays and determined the expression and activity of PXR through quantitative real-time polymerase chain reaction, western blotting and luciferase-based reporter gene assay. PXR knockdown approaches using shRNA-encoding vectors were applied to estimate the role of PXR for native MDR.

Results

Drug resistance ranged between 5.2 and 620 nM for paclitaxel, varied between 4.5 and 58 μM for cisplatin, and varied between 1.1 and 5,467 μM for 5-FU. Lack of PXR mRNA expression was mostly accompanied by the absence of mRNA expression of cytochrome P450 3A4 (CYP3A4) and P-glycoprotein (P-gp, ABCB1) expression. Neither mRNA nor protein expression of PXR correlated with drug resistance. However, PXR activity tended to correlate with IC₅₀ values of paclitaxel (p = 0.08). Knockdown of PXR in one of the cell lines had a slight but not significant impact on paclitaxel efficacy compared to scrambled sequence control. Surprisingly, only in two cell lines, PXR activity was increased by the well-known inductor rifampicin.

Conclusion

This study suggests a malfunctioning of PXR and thus a minor relevance for iatrogenic chemotherapy resistance in HNSCC.

Fanucchi M, Khuri FR (2004) Chemotherapy for recurrent or metastatic squamous cell carcinoma of the head and neck. Semin Oncol 31:809–815PubMedCrossRef

Gottesman MM, Fojo T, Bates SE (2002) Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer 2:48–58PubMedCrossRef

Theile D, Ketabi-Kiyanvash N, Herold-Mende C, Dyckhoff G, Efferth T, Bertholet V et al (2011) Evaluation of drug transporters’ significance for multidrug resistance in head and neck squamous cell carcinoma. Head Neck 33:959–968PubMedCrossRef

Blumberg B, Sabbagh W Jr, Juguilon H, Bolado J Jr, van Meter CM, Ong ES et al (1998) SXR, a novel steroid and xenobiotic-sensing nuclear receptor. Genes Dev 12:3195–3205PubMedCrossRef

Synold TW, Dussault I, Forman BM (2001) The orphan nuclear receptor SXR coordinately regulates drug metabolism and efflux. Nat Med 7:584–590PubMedCrossRef

Beijnen JH, Schellens JH (2004) Drug interactions in oncology. Lancet Oncol 5:489–496PubMedCrossRef

Desai PB, Nallani SC, Sane RS, Moore LB, Goodwin BJ, Buckley DJ et al (2002) Induction of cytochrome P450 3A4 in primary human hepatocytes and activation of the human pregnane X receptor by tamoxifen and 4-hydroxytamoxifen. Drug Metab Dispos 30:608–612PubMedCrossRef

Huang R, Murry DJ, Kolwankar D, Hall SD, Foster DR (2006) Vincristine transcriptional regulation of efflux drug transporters in carcinoma cell lines. Biochem Pharmacol 71:1695–1704PubMedCrossRef

Masuyama H, Hiramatsu Y, Kodama J, Kudo T (2003) Expression and potential roles of pregnane X receptor in endometrial cancer. J Clin Endocrinol Metab 88:4446–4454PubMedCrossRef

10.

Conde I, Lobo MV, Zamora J, Pérez J, González FJ, Alba E et al (2008) Human pregnane X receptor is expressed in breast carcinomas, potential heterodimers formation between hPXR and RXR-alpha. BMC Cancer 8:174–188PubMedCrossRef

11.

Takeyama D, Miki Y, Fujishima F, Suzuki T, Akahira J, Hata S (2010) Steroid and xenobiotic receptor in human esophageal squamous cell carcinoma: a potent prognostic factor. Cancer Sci 101:543–549PubMedCrossRef

12.

Rioja J, Bandrés E, Rosell Costa D, Rincón A, López I, Zudaire Bergera JJ et al (2011) Association of steroid and xenobiotic receptor (SXR) and multidrug resistance 1 (MDR1) gene expression with survival among patients with invasive bladder carcinoma. BJU Int 107:1833–1838PubMedCrossRef

13.

Ninck S, Reisser C, Dyckhoff G, Helmke B, Bauer H, Herold-Mende C (2003) Expression profiles of angiogenic growth factors in squamous cell carcinomas of the head and neck. Int J Cancer 106:34–44PubMedCrossRef

14.

Freier K, Hofele C, Gross M, Devens F, Dyckhoff G, Plinkert P et al (2010) Cytogenetic characterization of head and neck squamous cell carcinoma cell lines as model systems for the functional analyses of tumor-associated genes. J Oral Pathol Med 39:382–389PubMed

15.

Peters T, Lindenmaier H, Haefeli WE, Weiss J (2006) Interaction of the mitotic kinesin Eg5 inhibitor monastrol with P-glycoprotein. Naunyn Schmiedebergs Arch Pharmacol 372:291–299PubMedCrossRef

16.

Patel RD, Hollingshead BD, Omiecinski CJ, Perdew GH (2007) Aryl-hydrocarbon receptor activation regulates constitutive androstane receptor levels in murine and human liver. Hepatology 46:209–218PubMedCrossRef

17.

Theile D, Grebhardt S, Haefeli WE, Weiss J (2009) Involvement of drug transporters in the synergistic action of FOLFOX combination chemotherapy. Biochem Pharmacol 78:1366–1373PubMedCrossRef

18.

König SK, Herzog M, Theile D, Zembruski N, Haefeli WE, Weiss J (2010) Impact of drug transporters on cellular resistance towards saquinavir and darunavir. J Antimicrob Chemother 65:2319–2328PubMedCrossRef

19.

Albermann N, Schmitz-Winnenthal FH, Z’graggen K, Volk C, Hoffmann MM, Haefeli WE et al (2005) Expression of the drug transporters MDR1/ABCB1, MRP1/ABCC1, MRP2/ABCC2, BCRP/ABCG2, and PXR in peripheral blood mononuclear cells and their relationship with the expression in intestine and liver. Biochem Pharmacol 70:949–958PubMedCrossRef

20.

Jones SA, Moore LB, Shenk JL, Wisely GB, Hamilton GA, McKee DD et al (2000) The pregnane X receptor: a promiscuous xenobiotic receptor that has diverged during evolution. Mol Endocrinol 14:27–39PubMedCrossRef

21.

Cerveny L, Svecova L, Anzenbacherova E, Vrzal R, Staud F, Dvorak Z et al (2007) Valproic acid induces CYP3A4 and MDR1 gene expression by activation of constitutive androstane receptor and pregnane X receptor pathways. Drug Metab Dispos 35:1032–1041PubMedCrossRef

22.

Gu X, Ke S, Liu D, Sheng T, Thomas PE, Rabson AB et al (2006) Role of NF-kappaB in regulation of PXR-mediated gene expression: a mechanism for the suppression of cytochrome P-450 3A4 by proinflammatory agents. J Biol Chem 281:17882–17889PubMedCrossRef

23.

Weiss J, Theile D, Spalwisz A, Burhenne J, Riedel KD, Haefeli WE (2013) Influence of sildenafil and tadalafil on the enzyme- and transporter-inducing effects of bosentan and ambrisentan in LS180 cells. Biochem Pharmacol (in press)

24.

Masuyama H, Nakatsukasa H, Takamoto N, Hiramatsu Y (2007) Down-regulation of pregnane X receptor contributes to cell growth inhibition and apoptosis by anticancer agents in endometrial cancer cells. Mol Pharmacol 72:1045–1053PubMedCrossRef

25.

Scotto KW (2003) Transcriptional regulation of ABC drug transporters. Oncogene 22:7496–7511PubMedCrossRef

26.

Costea DE, Tsinkalovsky O, Vintermyr OK, Johannessen AC, Mackenzie IC (2006) Cancer stem cells—new and potentially important targets for the therapy of oral squamous cell carcinoma. Oral Dis 12:443–454PubMedCrossRef

27.

Diaz LA Jr, Williams RT, Wu J, Kinde I, Hecht JR, Berlin J et al (2012) The molecular evolution of acquired resistance to targeted EGFR blockade in colorectal cancers. Nature 486:537–540PubMed

28.

Blagosklonny MV (2002) STI-571 must select for drug-resistant cells but ‘no cell breathes fire out of its nostrils like a dragon’. Leukemia 16:570–572PubMedCrossRef

29.

Zuo C, Zhang H, Spencer HJ, Vural E, Suen JY, Schichman SA et al (2009) Increased microsatellite instability and epigenetic inactivation of the hMLH1 gene in head and neck squamous cell carcinoma. Otolaryngol Head Neck Surg 141:484–490PubMedCrossRef

30.

Bennett KL, Romigh T, Eng C (2009) AP-2alpha induces epigenetic silencing of tumor suppressive genes and microsatellite instability in head and neck squamous cell carcinoma. PLoS ONE 4:6931CrossRef

31.

Mensah-Osman EJ, Thomas DG, Tabb MM, Larios JM, Hughes DP, Giordano TJ et al (2007) Expression levels and activation of a PXR variant are directly related to drug resistance in osteosarcoma cell lines. Cancer 109:957–965PubMedCrossRef

32.

Takagi S, Nakajima M, Mohri T, Yokoi T (2008) Post-transcriptional regulation of human pregnane X receptor by micro-RNA affects the expression of cytochrome P450 3A4. J Biol Chem 283:9674–9680PubMedCrossRef

33.

Wang H, Venkatesh M, Li H, Goetz R, Mukherjee S, Biswas A et al (2011) Pregnane X receptor activation induces FGF19-dependent tumor aggressiveness in humans and mice. J Clin Invest 121:3220–3232PubMedCrossRef

34.

Piedade R, Schaeffeler E, Winter S, Asimus S, Schwab M, Ashton M et al (2012) PXR variants and artemisinin use in Vietnamese subjects: frequency distribution and impact on the interindividual variability of CYP3A induction by artemisinin. Antimicrob Agents Chemother 56:2153–2157PubMedCrossRef

35.

Lichti-Kaiser K, Xu C, Staudinger JL (2009) Cyclic AMP-dependent protein kinase signaling modulates pregnane x receptor activity in a species-specific manner. J Biol Chem 284:6639–6649PubMedCrossRef

Title: Minor role of pregnane-x-receptor for acquired multidrug resistance in head and neck squamous cell carcinoma in vitro
Authors: Juan Pablo Rigalli
Tasmin Reuter
Christel Herold-Mende
Gerhard Dyckhoff
Walter Emil Haefeli
Johanna Weiss
Dirk Theile
Publication date: 01-05-2013
Publisher: Springer-Verlag
Published in: Cancer Chemotherapy and Pharmacology / Issue 5/2013
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-013-2133-x

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

At a glance: The STEP trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 5/2013

A pilot feasibility study for cisplatin plus S-1 for the treatment for advanced or recurrent cervical cancer

A review of therapeutic challenges and achievements of methotrexate delivery systems for treatment of cancer and rheumatoid arthritis

Clinical implications of initial FDG-PET/CT in locally advanced rectal cancer treated with neoadjuvant chemoradiotherapy

Phase II randomized trial of carboplatin and gemcitabine with or without dexamethasone pre-treatment in patients with Stage IV non-small cell lung cancer

Combined targeting of FGFR2 and mTOR by ponatinib and ridaforolimus results in synergistic antitumor activity in FGFR2 mutant endometrial cancer models

Dodging a dogma: is treating beyond progression beneficial?

Keynote webinar | Spotlight on antibody–drug conjugates in cancer