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Differential gene expression in a paclitaxel-resistant clone of a head and neck cancer cell line

  • Head and Neck Oncology
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Abstract

The anti-neoplastic drug paclitaxel (taxol), which is known to block cells in the G2/M phase of the cell cycle through stabilization of microtubules, is meanwhile commonly used for chemotherapy of advanced head and neck cancer. Chemotherapy is primarily used in order to preserve laryngeal and/or pharyngeal structures. Although paclitaxel generally seems to be a powerful agent, it failed to reach a loco-regional tumor control in a sufficient percentage of patients. In order to investigate molecular resistance mechanisms, we have established a paclitaxel-resistant subline originating from the larynx carcinoma cell line HLaC79, which seemed to be partially dependent on taxol. The original and the descendant cell line were characterized by growth inhibition assays. We used western blotting and the cDNA subtraction (SSH) technique to identify genes differentially expressed in the taxol-resistant cell clone. cDNA subtraction revealed increased expression of six genes, including clathrin heavy chain, α3-tubulin, a neuroblastoma-specific Thymosin β, the ribosomal protein L7a, HLA-B associated transcript 3 and collagen IIIα1 in the taxol-resistant cell line. Furthermore, western blots showed an overexpression of MDR-1 in the taxol-resistant clone, while α- and β-tubulins and p48/IRF9 were expressed in equal amounts in both cell lines.

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References

  1. Bao L, Loda M, Janmey PA, Stewart R, Anand-Apte B, Zetter BR (1996) Thymosin beta 15: a novel regulator of tumor cell motility upregulated in metastatic prostate cancer. Nat Med 2:1322–1328

    PubMed  Google Scholar 

  2. Califano D, Monaco C, Santelli G, Giuliano A, Veronese ML, Berlingieri MT, de Franciscis V, Berger N, Trapasso F, Santoro M, Viglietto G, Fusco A (1998) Thymosin beta-10 gene overexpression correlated with the highly malignant neoplastic phenotype of transformed thyroid cells in vivo and in vitro. Cancer Res 58:823–828

    PubMed  Google Scholar 

  3. dos Remedios CG, Chabra D, CD, Kekic M, Dedova IV, Tsubakihara M, Berry DA, Nosworthy NJ (2003) Actin binding proteins: regulation of cytoskeletal microfilaments. Physiol Rev 83:433–473

    PubMed  Google Scholar 

  4. Giannakakou P, Sackett D, Fojo T (2000) Tubulin/microtubules: still a promising target for new chemotherapeutic agents. J Natl Cancer Inst. 92:182–183

    Google Scholar 

  5. Higgins CF, Callaghan R, Linton KJ, Rosenberg MF, Ford RC (1997) Structure of the multidrug resistance P-glycoprotein. Semin Cancer Biol 8:135–142

    PubMed  Google Scholar 

  6. Horwitz SB, Cohen D, Rao S, Ringel I, Shen HJ, Yang CP (1993) Taxol: mechanisms of action and resistance. J Natl Cancer Inst Monogr 15:55–61

    PubMed  Google Scholar 

  7. Ji P, Diederichs S, Wang W, Boing S, Metzger R, Schneider PM, Tidow N, Brandt B, Buerger H, Bulk E, Thomas M, Berdel WE, Serve H, Muller-Tidow C (2003) MALAT-1, a novel noncoding RNA, and thymosin beta4 predict metastasis and survival in early-stage non-small cell lung cancer. Oncogene 22:8031–8041

    PubMed  Google Scholar 

  8. Kauppila S, Stenback F, Risteli J, Jukkola A, Risteli L (1998) Aberrant type I and type III collagen gene expression in human breast cancer in vivo. J Pathol 186:262–268

    Article  PubMed  Google Scholar 

  9. Kelley DJ, Pavelic ZP, Gapany M, Stambrook P, Pavelic L, Gapany S, Gluckman JL (1993) Detection of P-glycoprotein in squamous cell carcinomas of the head and neck. Arch Otolaryngol Head Neck Surg 119:411–414

    PubMed  Google Scholar 

  10. Kim H, Barroso M, Samanta R, Greenberger L, Sztul E (1997) Experimentally induced changes in the endocytic traffic of P-glycoprotein alter drug resistance of cancer cells. Am J Physiol 273:C687–702

    PubMed  Google Scholar 

  11. Kroes RA, Jastrow A, McLone MG, Yamamoto H, Colley P, Kersey DS, Yong VW, Mkrdichian E, Cerullo L, Leestma J, Moskal JR (2000) The identification of novel therapeutic targets for the treatment of malignant brain tumors. Cancer Lett 156:191–198

    Article  PubMed  Google Scholar 

  12. Kyhse-Andersen J (1984) Electroblotting of multiple gels: a simple apparatus without tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Meth 10:203–210

    Article  PubMed  Google Scholar 

  13. Kyu-Ho Han E, Gehrke L, Tahir SK, Credo RB, Cherian SP, Sham H, Rosenberg SH, Ng S (2000) Modulation of drug resistance by α-tubulin in paclitaxel-resistant human lung cancer cell lines. Eur J Cancer 36:1565–1571

    PubMed  Google Scholar 

  14. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    PubMed  Google Scholar 

  15. Luker KE, Pica CM, Schreiber RD, Piwnica-Worms D (2001) Overexpression of IRF9 confers resistance to antimicrotubule agents in breast cancer cells. Cancer Res 61:6540–6547

    PubMed  Google Scholar 

  16. Manchen ST, Hubberstay AV (2001) Human Scythe contains a functional nuclear localization sequence and remains in the nucleus during staurosporine-induced apoptosis. Biochem Biophys Res Commun 287:1075–1082

    PubMed  Google Scholar 

  17. Martello LA, Verdier-Pinard P, Shen HJ, He L, Torres K, Orr GA, Horwitz SB (2003) Elevated levels of microtubule destabilizing factors in a Taxol-resistant/dependent A549 cell line with an α-tubulin mutation. Cancer Res 63:1207–1213

    PubMed  Google Scholar 

  18. McFarland DC (2000) Preparation of pure cell cultures by cloning. Meth Cell Sci 22:63–66

    Article  Google Scholar 

  19. Oien KA, Vass JK, Downie I, Fullarton G, Keith WN (2003) Profiling, comparison and validation of gene expression in gastric carcinoma and normal stomach. Oncogene 22:4287–4300

    PubMed  Google Scholar 

  20. Panda D, Miller HP, Banerjee A, Luduena RF, Wilson L (1994) Microtubule dynamics in vitro are regulated by the tubulin isotype composition. Proc Natl Acad Sci USA 91:11358–11362

    PubMed  Google Scholar 

  21. Pfreundner L, Hoppe F, Willner J, Preisler V, Bratengeier K, Hagen R, Helms J, Flentje M (2003) Induction chemotherapy with paclitaxel and cisplatin and CT-based 3D radiotherapy in patients with advanced laryngeal and hypopharyngeal carcinomas—a possibility for organ preservation. Radiother Oncol 68:163–170

    PubMed  Google Scholar 

  22. Schrijvers D, Vermorken JB (2000) Role of taxoids in head and neck cancer. Oncologist 5:199–208

    Article  PubMed  Google Scholar 

  23. Tannock IF, Lee CM, Tunggal JK, Cowan DS, Egorin MJ (2002) Limited penetration of anticancer drugs through tumor tissue: a potential cause of resistance of solid tumors to chemotherapy. Clin Cancer Res 8:878–884

    PubMed  Google Scholar 

  24. Tapper J, Kettunen E, El-Rifai W, Seppala M, Andersson LC, Knuutila S (2001) Changes in gene expression during progression of ovarian carcinoma. Cancer Genet Cytogenet 128:1–6

    PubMed  Google Scholar 

  25. Tseng CP, Cheng AJ, Chang JT, Tseng CH, Wang HM, Liao CT, Chen IH, Tseng KC (2002) Quantitative analysis of multidrug-resistance mdr1 gene expression in head and neck cancer by real-time RT-PCR. Jpn J Cancer Res 93:1230–1236

    PubMed  Google Scholar 

  26. Vaarala MH, Porvari KS, Kyllonen AP, Mustonen MV, Lukkarinen O, Vihko PT (1998) Several genes encoding ribosomal proteins are over-expressed in prostate-cancer cell lines: confirmation of L7a and L37 over-expression in prostate-cancer tissue samples. Int J Cancer 78:27–32

    Article  PubMed  Google Scholar 

  27. Verdier-Pinard P, Wang F, Martello L, Burd B, Orr GA, Horwitz SB (2003) Analysis of tubulin isotypes and mutations from taxol-resistant cells by combined isoelectrofocusing and mass spectrometry. Biochemistry 42:5349–5357

    Article  PubMed  Google Scholar 

  28. Wang WS, Chen PM, Hsiao HL, Wang HS, Liang WY, Su Y (2004) Overexpression of the thymosin beta-4 gene is associated with increased invasion of SW480 colon carcinoma cells and the distant metastasis of human colorectal carcinoma. Oncogene Jul 5 (Epub ahead of print)

  29. Wang Y, Cheong D, Chan S, Hooi SC (2000) Ribosomal protein L7a gene is up-regulated but not fused to the tyrosine kinase receptor as chimeric trk oncogene in human colorectal carcinoma. Int J Oncol 16:757–762

    PubMed  Google Scholar 

  30. Yeh JJ, Hsu WH, Wang JJ, Ho ST, Kao A (2003) Predicting chemotherapy response to paclitaxel-based therapy in advanced non-small-cell lung cancer with P-glycoprotein expression. Respiration 70:32–35

    Article  PubMed  Google Scholar 

  31. Yokoyama M, Nishi Y, Yoshii J, Okubo K, Matsubara K (1996) Identification and cloning of neuroblastoma-specific and nerve tissue-specific genes through compiled expression profiles. DNA Res 3:311–320

    Article  PubMed  Google Scholar 

  32. Zenner HP, Lehner W, Herrmann IF (1979) Establishment of carcinoma cell lines from larynx and submandibular gland. Arch Otorhinolaryngol 225:269–277

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Otorhinolaryngology, University of Würzburg, Josef-Schneider-Straße 11, 97080, Würzburg, Germany

    Marianne Schmidt, Gabriele Schler, Petra Gruensfelder & Florian Hoppe

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  1. Marianne Schmidt
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  2. Gabriele Schler
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  3. Petra Gruensfelder
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  4. Florian Hoppe
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Correspondence to Marianne Schmidt.

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Schmidt, M., Schler, G., Gruensfelder, P. et al. Differential gene expression in a paclitaxel-resistant clone of a head and neck cancer cell line. Eur Arch Otorhinolaryngol 263, 127–134 (2006). https://doi.org/10.1007/s00405-005-0936-z

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  • DOI: https://doi.org/10.1007/s00405-005-0936-z

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