Drug resistance to paclitaxel is not only associated with ABCB1 mRNA expression but also with drug accumulation in intracellular compartments in human lung cancer cell lines

  • Authors:
    • Masanori Shimomura
    • Takeshi Yaoi
    • Kyoko Itoh
    • Daishiro Kato
    • Kunihiko  Terauchi
    • Junichi Shimada
    • Shinji Fushiki
  • View Affiliations

  • Published online on: December 13, 2011     https://doi.org/10.3892/ijo.2011.1297
  • Pages: 995-1004
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

In order to clarify the mechanisms of resistance to paclitaxel in lung cancer, three human lung cancer cell lines which exhibit different sensitivity to paclitaxel were investigated from the following viewpoints: overexpression of ATP-binding cassette, sub-family B, member 1 (ABCB1), mutations on paclitaxel binding site of β-tubulin genes, quantity of polymerized tubulin and the intracellular localization of paclitaxel. ABCB1 expression was evaluated by real-time RT-PCR. No correlations were noted between the ABCB1 expression in the sensitive and resistant cell lines at the mRNA level. No mutations on the paclitaxel binding site of the β-tubulin genes were detected in either the resistant or sensitive cells. Live cell images obtained by confocal laser microscopy revealed that the resistant cell line, RERF-LC-KJ, had more accumulation of Oregon Green® 488 conjugated paclitaxel in the lysosomal and extra-lysosomal compartments of cytoplasm than other cell lines. The results obtained in this study indicated that the changes in the subcellular localization could contribute to the production of paclitaxel resistance in lung cancer cell lines. Further studies should be conducted to elucidate the molecular mechanisms that differentiate the intracellular localization of paclitaxel.

References

1 

Sawabata N, Asamura H, Goya T, et al: Japanese Lung Cancer Registry Study: first prospective enrollment of a large number of surgical and nonsurgical cases in 2002. J Thorac Oncol. 5:1369–1375. 2010. View Article : Google Scholar : PubMed/NCBI

2 

Pujol JL, Barlesi F and Daures JP: Should chemotherapy combinations for advanced non-small cell lung cancer be platinum-based? A meta-analysis of phase III randomized trials. Lung Cancer. 51:335–345. 2006. View Article : Google Scholar : PubMed/NCBI

3 

Ambudkar SV, Kimchi-Sarfaty C, Sauna ZE and Gottesman MM: P-glycoprotein: from genomics to mechanism. Oncogene. 22:7468–7485. 2003. View Article : Google Scholar : PubMed/NCBI

4 

Huang Y and Sadee W: Membrane transporters and channels in chemoresistance and sensitivity of tumor cells. Cancer Lett. 239:168–182. 2006. View Article : Google Scholar : PubMed/NCBI

5 

Lee JT Jr, Steelman LS and McCubrey JA: Phosphatidylinositol 3′-kinase activation leads to multidrug resistance protein-1 expression and subsequent chemoresistance in advanced prostate cancer cells. Cancer Res. 64:8397–8404. 2004.

6 

Giannakakou P, Sackett DL, Kang YK, et al: Paclitaxel-resistant human ovarian cancer cells have mutant beta-tubulins that exhibit impaired paclitaxel-driven polymerization. J Biol Chem. 272:17118–17125. 1997. View Article : Google Scholar

7 

Monzó M, Rosell R, Sánchez JJ, et al: Paclitaxel resistance in non-small-cell lung cancer associated with beta-tubulin gene mutations. J Clin Oncol. 17:1786–1793. 1999.PubMed/NCBI

8 

Hua XH, Genini D, Gussio R, et al: Biochemical genetic analysis of indanocine resistance in human leukemia. Cancer Res. 61:7248–7254. 2001.PubMed/NCBI

9 

Gonzalez-Garay ML, Chang L, Blade K, Menick DR and Cabral F: A beta-tubulin leucine cluster involved in microtubule assembly and paclitaxel resistance. J Biol Chem. 274:23875–23882. 1999. View Article : Google Scholar : PubMed/NCBI

10 

McGrogan BT, Gilmartin B, Carney DN and McCann A: Taxanes, microtubules and chemoresistant breast cancer. Biochim Biophys Acta. 1785:96–132. 2008.PubMed/NCBI

11 

Orr GA, Verdier-Pinard P, McDaid H and Horwitz SB: Mechanisms of Taxol resistance related to microtubules. Oncogene. 22:7280–7295. 2003. View Article : Google Scholar : PubMed/NCBI

12 

Berrieman HK, Lind MJ and Cawkwell L: Do beta-tubulin mutations have a role in resistance to chemotherapy? Lancet Oncol. 5:158–164. 2004. View Article : Google Scholar : PubMed/NCBI

13 

Kavallaris M, Tait AS, Walsh BJ, et al: Multiple microtubule alterations are associated with Vinca alkaloid resistance in human leukemia cells. Cancer Res. 61:5803–5809. 2001.PubMed/NCBI

14 

Hari M, Loganzo F, Annable T, et al: Paclitaxel-resistant cells have a mutation in the paclitaxel-binding region of beta-tubulin (Asp26Glu) and less stable microtubules. Mol Cancer Ther. 5:270–278. 2006. View Article : Google Scholar : PubMed/NCBI

15 

Yang H and Cabral F: Heightened sensitivity to paclitaxel in Class IVa beta-tubulin-transfected cells is lost as expression increases. J Biol Chem. 282:27058–27066. 2007. View Article : Google Scholar : PubMed/NCBI

16 

Aoki D, Oda Y, Hattori S, et al: Overexpression of class III beta-tubulin predicts good response to taxane-based chemotherapy in ovarian clear cell adenocarcinoma. Clin Cancer Res. 15:1473–1480. 2009. View Article : Google Scholar : PubMed/NCBI

17 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001.

18 

Sumizawa T, Chen ZS, Chuman Y, et al: Reversal of multidrug resistance-associated protein-mediated drug resistance by the pyridine analog PAK-104P. Mol Pharmacol. 51:399–405. 1997.PubMed/NCBI

19 

Shi Z, Tiwari AK, Shukla S, et al: Inhibiting the function of ABCB1 and ABCG2 by the EGFR tyrosine kinase inhibitor AG1478. Biochem Pharmacol. 77:781–793. 2009. View Article : Google Scholar : PubMed/NCBI

20 

Minotti AM, Barlow SB and Cabral F: Resistance to antimitotic drugs in Chinese hamster ovary cells correlates with changes in the level of polymerized tubulin. J Biol Chem. 266:3987–3994. 1991.PubMed/NCBI

21 

Westermann S and Weber K: Post-translational modifications regulate microtubule function. Nat Rev Mol Cell Biol. 4:938–947. 2003. View Article : Google Scholar : PubMed/NCBI

22 

Sasse R and Gull K: Tubulin post-translational modifications and the construction of microtubular organelles in Trypanosoma brucei. J Cell Sci. 90:577–589. 1988.PubMed/NCBI

23 

Belhoussine R, Morjani H, Sharonov S, Ploton D and Manfait M: Characterization of intracellular pH gradients in human multidrug-resistant tumor cells by means of scanning microspectrofluorometry and dual-emission-ratio probes. Int J Cancer. 81:81–89. 1999. View Article : Google Scholar : PubMed/NCBI

24 

Jiang LW, Maher VM, McCormick JJ and Schindler M: Alkalinization of the lysosomes is correlated with ras transformation of murine and human fibroblasts. J Biol Chem. 265:4775–4777. 1990.PubMed/NCBI

25 

Altan N, Chen Y, Schindler M and Simon SM: Defective acidification in human breast tumor cells and implications for chemotherapy. J Exp Med. 187:1583–1598. 1998. View Article : Google Scholar : PubMed/NCBI

26 

Gong Y, Duvvuri M and Krise JP: Separate roles for the Golgi apparatus and lysosomes in the sequestration of drugs in the multidrug-resistant human leukemic cell line HL-60. J Biol Chem. 278:50234–50239. 2003. View Article : Google Scholar : PubMed/NCBI

27 

Kokkonen N, Rivinoja A, Kauppila A, Suokas M, Kellokumpu I and Kellokumpu S: Defective acidification of intracellular organelles results in aberrant secretion of cathepsin D in cancer cells. J Biol Chem. 279:39982–39988. 2004. View Article : Google Scholar : PubMed/NCBI

28 

Alvarez M, Paull K, Monks A, et al: Generation of a drug resistance profile by quantitation of mdr-1/P-glycoprotein in the cell lines of the National Cancer Institute Anticancer Drug Screen. J Clin Invest. 95:2205–2214. 1995. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

April 2012
Volume 40 Issue 4

Print ISSN: 1019-6439
Online ISSN:1791-2423

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Shimomura, M., Yaoi, T., Itoh, K., Kato, D., Terauchi, K., Shimada, J., & Fushiki, S. (2012). Drug resistance to paclitaxel is not only associated with ABCB1 mRNA expression but also with drug accumulation in intracellular compartments in human lung cancer cell lines. International Journal of Oncology, 40, 995-1004. https://doi.org/10.3892/ijo.2011.1297
MLA
Shimomura, M., Yaoi, T., Itoh, K., Kato, D., Terauchi, K., Shimada, J., Fushiki, S."Drug resistance to paclitaxel is not only associated with ABCB1 mRNA expression but also with drug accumulation in intracellular compartments in human lung cancer cell lines". International Journal of Oncology 40.4 (2012): 995-1004.
Chicago
Shimomura, M., Yaoi, T., Itoh, K., Kato, D., Terauchi, K., Shimada, J., Fushiki, S."Drug resistance to paclitaxel is not only associated with ABCB1 mRNA expression but also with drug accumulation in intracellular compartments in human lung cancer cell lines". International Journal of Oncology 40, no. 4 (2012): 995-1004. https://doi.org/10.3892/ijo.2011.1297