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Mechanistic Analysis of Taxol-induced Multidrug Resistance in an Ovarian Cancer Cell Line

  • Wang, Ning-Ning (Department of Gynecological Oncology Surgery, Jiangsu Cancer Hospital & Institute, Nanjing Medical University) ;
  • Zhao, Li-Jun (Department of Gynecological Oncology Surgery, Jiangsu Cancer Hospital & Institute, Nanjing Medical University) ;
  • Wu, Li-Nan (Department of Gynecological Oncology Surgery, Jiangsu Cancer Hospital & Institute, Nanjing Medical University) ;
  • He, Ming-Feng (Nanjing University of Technology School of Pharmaceutical Science) ;
  • Qu, Jun-Wei (Department of Gynecological Oncology Surgery, Jiangsu Cancer Hospital & Institute, Nanjing Medical University) ;
  • Zhao, Yi-Bing (Department of Gynecological Oncology Surgery, Jiangsu Cancer Hospital & Institute, Nanjing Medical University) ;
  • Zhao, Wan-Zhou (Sino-EU Biomedical Innovation Center (SEBIC), OG Pharma Corporation) ;
  • Li, Jie-Shou (Department of General Surgery, Nanjing General Hospital of Nanjing Military Command) ;
  • Wang, Jin-Hua (Department of Gynecological Oncology Surgery, Jiangsu Cancer Hospital & Institute, Nanjing Medical University)
  • Published : 2013.09.30

Abstract

Objectives: To establish a taxol-resistant cell line of human ovarian carcinoma (A2780/Taxol) and investigate its biological features. Methods: The drug-resistant cell line (A2780/Taxol) was established by continuous stepwise selection with increasing concentrations of Taxol. Cell morphology was assessed by microscopy and growth curves were generated with in vitro and in vivo tumor xenograft models. With rhodamine123 (Rh123) assays, cell cycle distribution and the apoptotic rate were analyzed by flow cytometry (FCM). Drug resistance-related and signal associated proteins, including P-gp, MRPs, caveolin-1, PKC-${\alpha}$, Akt, ERK1/2, were detected by Western blotting. Results: A2780/Taxol cells were established with stable resistance to taxol. The drug resistance index (RI) was 430.7. Cross-resistance to other drugs was also shown, but there was no significant change to radioresistance. Compared with parental cells, A2780/Taxol cells were significantly heteromorphous, with a significant delay in population doubling time and reduced uptake of Rh123 (p<0.01). In vivo, tumor take by A2780 cells was 80%, and tumor volume increased gradually. In contrast, with A2780/Taxol cells in xenograft models there was no tumor development. FCM analysis revealed that A2780/Taxol cells had a higher percentage of G0/G1 and lower S phase, but no changes of G2 phase and the apoptosis rate. Expression of P-gp, MRP1, MRP2, BCRP, LRP, caveolin-1, PKC-${\alpha}$, Phospho-ERK1/2 and Phospho-JNK protein was significantly up-regulated, while Akt and p38 MARK protein expression was not changed in A2780/Taxol cells. Conclusion: The A2780/Taxol cell line is an ideal model to investigate the mechanism of muti-drug resistance related to overexpression of drug-resistance associated proteins and activation of the PKC-${\alpha}/ERK$ (JNK) signaling pathway.

Keywords

Ovarian carcinoma;Taxol;Multidrug resistance;signaling pathway

References

  1. Cantley LC (2002). The phosphoinositide 3-kinase pathway. Science, 296, 1655-7. https://doi.org/10.1126/science.296.5573.1655
  2. Chauhan D, Tian Z, Nicholson B, et al (2012). A small molecule inhibitor of ubiquitin-specific protease-7 induces apoptosis in multiple myeloma cells and overcomes bortezomib resistance. Cancer Cell, 22, 345-58. https://doi.org/10.1016/j.ccr.2012.08.007
  3. Di Michele M, Della Corte A, Cicchillitti L, et al (2009). A proteomic approach to paclitaxel chemoresistance in ovarian cancer cell lines. Biochim Biophys Acta, 1794, 225-36. https://doi.org/10.1016/j.bbapap.2008.09.017
  4. Doddareddy MR, Rawling T, Ammit AJ(2012).Targeting mitogen-activated protein kinase phosphatase-1 (MKP-1): structure-based design of MKP-1 inhibitors and upregulators. Curr Med Chem, 19, 163-73. https://doi.org/10.2174/092986712803414196
  5. Downward J (2003). Targeting RAS signaling pathways in cancer therapy. Nat Rev Cancer, 3, 11-22. https://doi.org/10.1038/nrc969
  6. Hiss D (2012). Optimizing molecular-targeted therapies in ovarian cancer: the renewed surge of interest in ovarian cancer biomarkers and cell signaling pathways. J Oncol, 2012, 737981.
  7. Liu HZ, Yu C, Yang Z, et al (2011). Tubeimoside I sensitizes cisplatin in cisplatin-resistant human ovarian cancer cells (A2780/DDP) through down-regulation of ERK and upregulation of p38 signaling pathways. Mol Med Report, 4, 985-92.
  8. Hoshino R, Chatani Y, Yamori T, et al (1999). Constitutive activation of the 41/43-kDa mitogen-activated protein kinase signaling pathway in human tumors. Oncogene, 18, 813-22. https://doi.org/10.1038/sj.onc.1202367
  9. Kim H, Park GS, Lee JE, et al (2013). A leukotriene B4 receptor-2 is associated with paclitaxel resistance in MCF-7/ DOX breast cancer cells. Br J Cancer, 109, 351-9. https://doi.org/10.1038/bjc.2013.333
  10. LeBlanc R, Catley LP, Hideshima T, et al(2002). Proteasome inhibitor PS-341 inhibits human myeloma cell growth in vivo and prolongs survival in a murine model. Cancer Res, 62, 4996-5000.
  11. Mackay HJ, Twelves CJ (2003). Protein kinase C: a target for anticancer drugs? Endocr Relat Cancer, 10, 389-96. https://doi.org/10.1677/erc.0.0100389
  12. McCubrey JA, Steelman LS, Chappell WH, et al (2012). Ras/ Raf/MEK/ERK and PI3K/PTEN/Akt/mTOR cascade inhibitors: how mutations can result in therapy resistance and how to overcome resistance. Oncotarget, 3, 1068-111.
  13. Navolanic PM, Steelman LS, McCubrey JA (2003). EGFR family signaling and its association with breast cancer development and resistance to chemotherapy (Review). Int J Oncol, 22, 237-52.
  14. Paraiso KH, Smalley KS (2013).Fibroblast-mediated drug resistance in cancer. Biochem Pharmacol, 85, 1033-41. https://doi.org/10.1016/j.bcp.2013.01.018
  15. Perez-Tomas R(2006). Multidrug resistance: retrospect and prospects in anti- cancer drug treatment. Curr Med Chem, 13, 1859-76. https://doi.org/10.2174/092986706777585077
  16. Tanaka Y, Gavrielides MV, Mitsuuchi Y, et al (2003). Protein kinase C promotes apoptosis in LNCaP prostate cancer cells through activation of p38 MAPK and inhibition of the Akt survival pathway. J Biol Chem, 278, 33753-62. https://doi.org/10.1074/jbc.M303313200
  17. Salem AF, Bonuccelli G, Bevilacqua G, et al (2011). Caveolin-1 promotes pancreatic cancer cell differentiation and restores membranous E-cadherin via suppression of the epithelialmesenchymal transition. Cell Cycle, 10, 3692-700. https://doi.org/10.4161/cc.10.21.17895
  18. Sui H, Fan ZZ, Li Q (2012).Signal transduction pathways and transcriptional mechanisms of ABCB1/Pgp-mediated multiple drug resistance in human cancer cells. J Int Med Res, 40, 426-35. https://doi.org/10.1177/147323001204000204
  19. Szakacs G, Paterson J, Ludwig JA, et al (2006). Targeting multidrug resistance in cancer. Nat Rev Drug Discov, 5, 219-34. https://doi.org/10.1038/nrd1984
  20. Wang JH, Zhao WZ, Chen XX, et al (2010). Establishment and biological characterization of Cisplatin-resistant cell line OV1228/Taxol of ovarian carcinoma. Chin J Cancer Prev Treat, 22, 281-4.
  21. Wee S, Wiederschain D, Maira SM, et al (2008). PTEN-deficient cancers depend on PIK3CB. Proc Natl Acad Sci U S A, 105, 13057-62. https://doi.org/10.1073/pnas.0802655105
  22. Wen J, Zheng B, Hu Y, et al (2009). Establishment and biological analysis of the EC109/CDDP multidrug-resistant esophageal squamous cell carcinoma cell line. Oncol Rep, 22, 65-71.
  23. Wu DL, Huang F, Lu HZ(2003). Drug-resistant proteins in breast cancer: recent progress in multidrug resistance [J]. Chin J Cancer, 22, 441-4. [in Chinese]
  24. Yan XD, Li M, Yuan Y, et al (2007). Biological comparison of ovarian cancer resistant cell lines to cisplatin and Taxol by two different administrations. Oncol Rep, 17, 1163-9.
  25. Yang SH, Sharrocks AD, Whitmarsh AJ (2013). MAP kinase signalling cascades and transcriptional regulation. Gene, 513, 1-13. https://doi.org/10.1016/j.gene.2012.10.033
  26. Zhao LJ, Xu H, Qu JW (2012). Modulation of drug resistance in ovarian cancer cells by inhibition of protein kinase C-alpha (PKC-$\alpha$) with small interference RNA (siRNA) agents. Asian Pac J Cancer Prev, 13, 3631-6. https://doi.org/10.7314/APJCP.2012.13.8.3631
  27. Zhe C, Li-Juan W, Ming Hui W, et al (2011). Mechanism governing reversal of multidrug resistance in human breast carcinoma cells by chelerythrine. Zhongguo Yi Xue Ke Xue Yuan Xue Bao, 33, 45-50.

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