DOI QR코드

DOI QR Code

Establishment of Paclitaxel-resistant Breast Cancer Cell Line and Nude Mice Models, and Underlying Multidrug Resistance Mechanisms in Vitro and in Vivo

  • Chen, Si-Ying (Department of Pharmacy, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University) ;
  • Hu, Sa-Sa (Department of Pharmacy, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University) ;
  • Dong, Qian (Qilu Pharmaceutical Co., Ltd.) ;
  • Cai, Jiang-Xia (Department of Pharmacy, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University) ;
  • Zhang, Wei-Peng (Department of Pharmacy, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University) ;
  • Sun, Jin-Yao (Department of Pharmacy, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University) ;
  • Wang, Tao-Tao (Department of Pharmacy, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University) ;
  • Xie, Jiao (Department of Pharmacy, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University) ;
  • He, Hai-Rong (Department of Pharmacy, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University) ;
  • Xing, Jian-Feng (Department of Pharmacy, College of Medicine, Xi'an Jiaotong University) ;
  • Lu, Jun (Department of Pharmacy, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University) ;
  • Dong, Ya-Lin (Department of Pharmacy, the First Affiliated Hospital of Medical College, Xi'an Jiaotong University)
  • Published : 2013.10.30

Abstract

Background: Breast cancer is a common malignant tumor which affects health of women and multidrug resistance (MDR) is one of the main factors leading to failure of chemotherapy. This study was conducted to establish paclitaxel-resistant breast cancer cell line and nude mice models to explore underlying mechanisms of MDR. Methods: The breast cancer drug-sensitive cell line MCF-7 (MCF-7/S) was exposed in stepwise escalating paclitaxel (TAX) to induce a resistant cell line MCF-7/TAX. Cell sensitivity to drugs and growth curves were measured by MTT assay. Changes of cell morphology and ultrastructure were examined by optical and electron microscopy. The cell cycle distribution was determined by flow cytometry. Furthermore, expression of proteins related to breast cancer occurrence and MDR was tested by immunocytochemistry. In Vivo, nude mice were injected with MCF-7/S and MCF-7/TAX cells and weights and tumor sizes were observed after paclitaxel treatment. In addition, proteins involved breast cancer and MDR were detected by immunohistochemistry. Results: Compared to MCF-7/S, MCF-7/TAX cells had a higher resistance to paclitaxel, cross-resistance and prolonged doubling time. Moreover, MCF-7/TAX showed obvious alterations of ultrastructure. Estrogen receptor (ER) expression was low in drug resistant cells and tumors while expression of human epidermal growth factor receptor 2 (HER2) and Ki-67 was up-regulated. P-glycoprotein (P-gp), lung resistance-related protein (LRP) and glutathione-S-transferase-${\pi}$ (GST-${\pi}$) involved in the MDR phenotype of resistant cells and tumors were all overexpressed. Conclusion: The underlying MDR mechanism of breast cancer may involve increased expression of P-gp, LRP and GST-${\pi}$.

Keywords

Breast cancer;paclitaxel;chemotherapy;multidrug resistance

Acknowledgement

Supported by : National Natural Science Foundation of China

References

  1. Zhou HL, Zheng YJ, Cheng XZ, et al (2013). Intercellular transfer of P-glycoprotein from the drug resistant human bladder cancer cell line BIU-87 does not require cell-to-cell contact. J Urol, 190, 1069-75. https://doi.org/10.1016/j.juro.2013.04.053
  2. Zhang W, Ding W, Chen Y, et al (2011). Up-regulation of breast cancer resistance protein plays a role in HER2-mediated chemoresistance through PI3K/Akt and nuclear factor-kappa B signaling pathways in MCF7 breast cancer cells. Acta Biochim Biophys Sin (Shanghai), 43, 647-53. https://doi.org/10.1093/abbs/gmr050
  3. Zhao YX, Liang WQ, Wang Y, Liu DX (2011). Cationic submicron emulsions overcome multidrug resistance in SGC7901/VCR cells. Pharmazie, 66, 130-5.
  4. Urruticoechea A, Smith IE, Dowsett M (2005). Proliferation marker Ki-67 in early breast cancer. J Clin Oncol, 23, 7212-20. https://doi.org/10.1200/JCO.2005.07.501
  5. Yu ST, Chen TM, Chern JW, Tseng SY, Chen YH (2009). Downregulation of GSTpi expression by tryptanthrin contributing to sensitization of doxorubicin-resistant MCF-7 cells through c-jun NH2-terminal kinase-mediated apoptosis. Anticancer Drugs, 20, 382-8. https://doi.org/10.1097/CAD.0b013e32832a2cd4
  6. Arai T, Miyoshi Y, Kim S J, et al (2008). Association of GSTP1 expression with resistance to docetaxel and paclitaxel in human breast cancers. Eur J Surg Oncol, 34, 734-8. https://doi.org/10.1016/j.ejso.2007.07.008
  7. Arteaga CL, Moulder SL, Yakes FM (2002). HER (erbB) tyrosine kinase inhibitors in the treatment of breast cancer. Semin Oncol, 29, 4-10.
  8. Baguley BC (2010). Multiple Drug Resistance Mechanisms in Cancer. Mol Biotechnol, 46, 308-16. https://doi.org/10.1007/s12033-010-9321-2
  9. Ballerini S, Bellincampi L, Bernardini S, et al (2003). Analysis of GSTP1-1 polymorphism using real-time polymerase chain reaction. Clinica Chimica Acta, 329, 127-32. https://doi.org/10.1016/S0009-8981(02)00422-9
  10. Dizdarevic S, Peters AM (2011). Imaging of multidrug resistance in cancer. Cancer Imaging, 11, 1-8. https://doi.org/10.1102/1470-7330.2011.0001
  11. Bedard PL, Di Leo A, Piccart-Gebhart MJ (2010). Taxanes: optimizing adjuvant chemotherapy for early-stage breast cancer. Nat Rev Clin Oncol, 7, 22-36. https://doi.org/10.1038/nrclinonc.2009.186
  12. Bhoo-Pathy N, Yip CH, Hartman M, et al (2013). Breast cancer research in Asia: adopt or adapt Western knowledge? Eur J Cancer, 49, 703-9. https://doi.org/10.1016/j.ejca.2012.09.014
  13. Coughlin SS, Ekwueme DU (2009). Breast cancer as a global health concern. Cancer Epidemiol, 33, 315-8. https://doi.org/10.1016/j.canep.2009.10.003
  14. Fromm MF (2002). The influence of MDR1 polymorphisms on P-glycoprotein expression and function in humans. Adv Drug Deliv Rev, 54, 1295-310. https://doi.org/10.1016/S0169-409X(02)00064-9
  15. Gu W, Fang FF, Li B, Cheng BB, Ling CQ (2012).Characterization and resistance mechanisms of a 5-fluorouracil- resistant hepatocellular carcinoma cell line. Asian Pac J Cancer Prev, 13, 4807-14. https://doi.org/10.7314/APJCP.2012.13.9.4807
  16. Kaira K, Takahashi T, Murakami H, et al (2013). The role of betaIII-tubulin in non-small cell lung cancer patients treated by taxane-based chemotherapy. Int J Clin Oncol, 18, 371-9. https://doi.org/10.1007/s10147-012-0386-8
  17. Kerr EH, Frederick PJ, Egger ME, et al (2013). Lung resistance-related protein (LRP) expression in malignant ascitic cells as a prognostic marker for advanced ovarian serous carcinoma. Ann Surg Oncol, 20, 3059-65. https://doi.org/10.1245/s10434-013-2878-9
  18. Lloyd HH (1974). Combination chemotherapy: considerations for design and analysis. Cancer Chemother Rep 2, 4, 157-65.
  19. Lu L, Zhou D, Jiang X, et al (2012). Loss of E-cadherin in multidrug resistant breast cancer cell line MCF-7/Adr: possible implication in the enhanced invasive ability. Eur Rev Med Pharmacol Sci, 16, 1271-9.
  20. Murray S, Briasoulis E, Linardou H, Bafaloukos D, Papadimitriou C (2012). Taxane resistance in breast cancer: mechanisms, predictive biomarkers and circumvention strategies. Cancer Treat Rev, 38, 890-903. https://doi.org/10.1016/j.ctrv.2012.02.011
  21. Podolski-Renic A, Andelkovic T, Bankovic J, et al (2011). The role of paclitaxel in the development and treatment of multidrug resistant cancer cell lines. Biomed Pharmacother, 65, 345-53. https://doi.org/10.1016/j.biopha.2011.04.015
  22. Saeki T, Tsuruo T, Sato W, Nishikawsa K (2005). Drug resistance in chemotherapy for breast cancer. Cancer Chemother Pharmacol, 56, 84-9. https://doi.org/10.1007/s00280-005-0106-4
  23. Saloustros E, Mavroudis D, Georgoulias V (2008). Paclitaxel and docetaxel in the treatment of breast cancer. Expert Opin Pharmacother, 9, 2603-16. https://doi.org/10.1517/14656566.9.15.2603
  24. Siegel R, DeSantis C, Virgo K, et al (2012). Cancer treatment and survivorship statistics, 2012. CA Cancer J Clin, 62, 220-41. https://doi.org/10.3322/caac.21149
  25. Slamon DJ, Leyland-Jones B, Shak S, et al (2001). Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. N Engl J Med, 344, 783-92. https://doi.org/10.1056/NEJM200103153441101
  26. Smoter M, Bodnar L, Duchnowska R, et al (2011). The role of Tau protein in resistance to paclitaxel. Cancer Chemother Pharmacol, 68, 553-7. https://doi.org/10.1007/s00280-011-1696-7
  27. Snow K, Judd W (1991). Characterisation of adriamycin- and amsacrine-resistant human leukaemic T cell lines. Br J Cancer, 63, 17-28. https://doi.org/10.1038/bjc.1991.7
  28. Song YN, Guo XL, Zheng BB, et al (2011). Ligustrazine derivate DLJ14 reduces multidrug resistance of K562/A02 cells by modulating GSTpi activity. Toxicology In Vitro, 25, 937-43. https://doi.org/10.1016/j.tiv.2011.03.002

Cited by

  1. Paclitaxel resistance in MCF-7/PTX cells is reversed by paeonol through suppression of the SET/phosphatidylinositol 3-kinase/Akt pathway vol.12, pp.1, 2012, https://doi.org/10.3892/mmr.2015.3468
  2. Acquisition of epithelial–mesenchymal transition is associated with Skp2 expression in paclitaxel-resistant breast cancer cells vol.110, pp.8, 2014, https://doi.org/10.1038/bjc.2014.136
  3. Establishment and Partial Characterization of an Epirubicin-Resistant Gastric Cancer Cell Line with Upregulated ABCB1 vol.15, pp.16, 2014, https://doi.org/10.7314/APJCP.2014.15.16.6849
  4. Over-Expression of Beclin-1 Facilitates Acquired Resistance to Histone Deacetylase Inhibitor-Induced Apoptosis vol.15, pp.18, 2014, https://doi.org/10.7314/APJCP.2014.15.18.7913
  5. EGF Reverses Multi-drug Resistance via the p-ERK Pathway in HepG2/ADM and SMMC7721/ADM Hepatocellular Carcinoma Models vol.15, pp.6, 2014, https://doi.org/10.7314/APJCP.2014.15.6.2619
  6. Effects of Ribosomal Protein L39-L on the Drug Resistance Mechanisms of Lung Cancer A549 Cells vol.15, pp.7, 2014, https://doi.org/10.7314/APJCP.2014.15.7.3093
  7. Evodiamine Synergizes with Doxorubicin in the Treatment of Chemoresistant Human Breast Cancer without Inhibiting P-Glycoprotein vol.9, pp.5, 2014, https://doi.org/10.1371/journal.pone.0097512
  8. Establishment and characterization of gemcitabine-resistant human cholangiocarcinoma cell lines with multidrug resistance and enhanced invasiveness vol.47, pp.1, 2015, https://doi.org/10.3892/ijo.2015.3019
  9. Evaluation of the drug sensitivity and expression of 16 drug resistance-related genes in canine histiocytic sarcoma cell lines vol.77, pp.6, 2015, https://doi.org/10.1292/jvms.14-0415
  10. Hurdles in selection process of nanodelivery systems for multidrug-resistant cancer vol.142, pp.10, 2016, https://doi.org/10.1007/s00432-016-2167-7
  11. SET protein overexpression contributes to paclitaxel resistance in MCF-7/S cells through PI3K/Akt pathway vol.25, pp.3, 2017, https://doi.org/10.1080/1061186X.2016.1245307
  12. CLDN6 promotes chemoresistance through GSTP1 in human breast cancer vol.36, pp.1, 2017, https://doi.org/10.1186/s13046-017-0627-9
  13. Preclinical studies for the combination of paclitaxel and curcumin in cancer therapy vol.37, pp.6, 2017, https://doi.org/10.3892/or.2017.5593
  14. SB-T-121205, a next-generation taxane, enhances apoptosis and inhibits migration/invasion in MCF-7/PTX cells vol.50, pp.3, 2017, https://doi.org/10.3892/ijo.2017.3871
  15. A Smart Paclitaxel-Disulfiram Nanococrystals for Efficient MDR Reversal and Enhanced Apoptosis vol.35, pp.4, 2018, https://doi.org/10.1007/s11095-018-2370-0