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Differential Expression of HSP90β in MDA-MB-231 and MCF-7 Cell Lines after Treatment with Doxorubicin

Jokar, Fereshte;Mahabadi, Javad Amini;Salimian, Morteza;Taherian, Aliakbar;Hayat, Seyyed Mohammad Gheibi;Sahebkar, Amirhossein;Atlasi, Mohammad Ali

  • Received : 2018.04.21
  • Accepted : 2019.02.11
  • Published : 2019.03.31

Abstract

Background: Breast cancer is a complex, heterogeneous disease and one of the most common malignancies in women worldwide. The efficacy of chemotherapy as an important breast cancer treatment option has been severely limited because of the inherent or acquired resistance of cancer cells. The molecular chaperone heat shock protein 90 (HSP90) upregulated in response to cellular stress is required for functions such as conformational maturation, activation and stability in more than 200 client proteins, mostly of the signaling type. In this study, the expression of HSP90 isoforms including $HSP90{\alpha}$ and $HSP90{\beta}$ in breast cancer cell lines before and after treatment with doxorubicin (DOX) was assessed. Material and Methods: The cell cytotoxicity of DOX in MDA-MB-231 and MCF-7 cell lines was determined using the MTT assay. immunofluorescence and western blotting techniques were used to determine the expression of $HSP90{\beta}$ in the cell lines before and after DOX treatment. Immunofluorescence was also conducted to ascertain the expression of $HSP90{\alpha}$. Results: The MTT assay results showed that the MDA-MB-231 cells ($IC_{50}=14.521{\mu}M$) were more sensitive than the MCF-7 cells ($IC_{50}=16.3315{\mu}M$) to DOX. The immunofluorescence results indicated that the expression of $HSP90{\alpha}$ in both cell lines decreased after exposure to DOX. The western blot and immunofluorescence analyses showed that $HSP90{\beta}$ expression decreased in the MCF-7 cells but increased in the MDA-MB-231 cells after DOX treatment. Conclusion: The obtained results suggested that $HSP90{\alpha}$ and $HSP90{\beta}$ expression levels were reduced in the MCF-7 cells after exposure to DOX. In the MDA-MB-231 cells, $HSP90{\alpha}$ expression was reduced while $HSP90{\beta}$ was found to be overexpressed following DOX treatment.

Keywords

$HSP90{\alpha}$;$HSP90{\beta}$;MCF-7;MDA-MB-231;breast cancer;heat shock protein

References

  1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2015. CA: a cancer journal for clinicians. 2015;65(1):5-29. https://doi.org/10.3322/caac.21254
  2. Veronesi U, et al. Rethinking TNM: a breast cancer classification to guide to treatment and facilitate research. The breast journal. 2009;15(3):291-295. https://doi.org/10.1111/j.1524-4741.2009.00719.x
  3. Longley D, Johnston P. Molecular mechanisms of drug resistance. The Journal of pathology. 2005;205(2):275-292. https://doi.org/10.1002/path.1706
  4. Aas T, et al. Specific P53 mutations are associated with de novo resistance to doxorubicin in breast cancer patients. Nature medicine. 1996;2(7):811-814. https://doi.org/10.1038/nm0796-811
  5. Harris AL, Hochhauser D. Mechanisms of multidrug resistance in cancer treatment. Acta Oncologica. 1992;31(2):205-213. https://doi.org/10.3109/02841869209088904
  6. Jacks T, Weinberg RA. Taking the study of cancer cell survival to a new dimension. Cell. 2002;111(7):923-925. https://doi.org/10.1016/S0092-8674(02)01229-1
  7. Joly AL, et al. Dual role of heat shock proteins as regulators of apoptosis and innate immunity. Journal of Innate Immunity. 2009;2(3):238-247.
  8. Echeverria PC, et al. Detection of changes in gene regulatory patterns, elicited by perturbations of the Hsp90 molecular chaperone complex, by visualizing multiple experiments with an animation. BioData Min. 2011;4(1):15. https://doi.org/10.1186/1756-0381-4-15
  9. Hickey E, et al. Sequence and regulation of a gene encoding a human 89-kilodalton heat shock protein. Mol Cell Biol. 1989;9(6):2615-26. https://doi.org/10.1128/MCB.9.6.2615
  10. Rebbe NF, et al. Nucleotide sequence and regulation of a human 90-kDa heat shock protein gene. Journal of Biological Chemistry. 1989;264(25):15006-15011.
  11. Neve RM, et al. A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer cell. 2006;10(6):515-527. https://doi.org/10.1016/j.ccr.2006.10.008
  12. Chin K, et al. Genomic and transcriptional aberrations linked to breast cancer pathophysiologies. Cancer cell. 2006;10(6):529-541. https://doi.org/10.1016/j.ccr.2006.10.009
  13. Hollestelle A, et al. Distinct gene mutation profiles among luminal-type and basal-type breast cancer cell lines. Breast cancer research and treatment. 2010;121(1):53-64. https://doi.org/10.1007/s10549-009-0460-8
  14. Kao J, et al. Molecular profiling of breast cancer cell lines defines relevant tumor models and provides a resource for cancer gene discovery. PloS one. 2009;4(7):e6146. https://doi.org/10.1371/journal.pone.0006146
  15. Lacroix M, Leclercq G. Relevance of breast cancer cell lines as models for breast tumours: an update. Breast cancer research and treatment. 2004;83(3):249-289. https://doi.org/10.1023/B:BREA.0000014042.54925.cc
  16. Vargo-Gogola T, Rosen JM. Modelling breast cancer: one size does not fit all. Nature Reviews Cancer. 2007;7(9):659-672. https://doi.org/10.1038/nrc2193
  17. Soule H, et al. A human cell line from a pleural effusion derived from a breast carcinoma. Journal of the National Cancer Institute. 1973;51(5):1409-1416. https://doi.org/10.1093/jnci/51.5.1409
  18. Wood AJ, Osborne CK. Tamoxifen in the treatment of breast cancer. New England Journal of Medicine. 1998;339(22):1609-1618. https://doi.org/10.1056/NEJM199811263392207
  19. Fabbro D, et al. Epidermal growth factor binding and protein kinase C activities in human breast cancer cell lines: possible quantitative relationship. Cancer research. 1986;46(6):2720-2725.
  20. Prat A, et al. Phenotypic and molecular characterization of the claudin-low intrinsic subtype of breast cancer. Breast Cancer Res. 2010;12(5):R68. https://doi.org/10.1186/bcr2635
  21. Heiser LM, et al. Subtype and pathway specific responses to anticancer compounds in breast cancer. Proceedings of the National Academy of Sciences. 2012;109(8):2724-2729. https://doi.org/10.1073/pnas.1018854108
  22. Visagie M, Mqoco T, Joubert A. Sulphamoylated estradiol analogue induces antiproliferative activity and apoptosis in breast cell lines. Cellular and Molecular Biology Letters. 2012;17(4):549-558.
  23. Georgakis GV, et al. The HSP90 inhibitor 17-AAG synergizes with doxorubicin and U0126 in anaplastic large cell lymphoma irrespective of ALK expression. Exp Hematol. 2006;34(12):1670-9. https://doi.org/10.1016/j.exphem.2006.07.002
  24. Soti C, et al. Heat shock proteins as emerging therapeutic targets. British journal of pharmacology. 2005;146(6):769-780. https://doi.org/10.1038/sj.bjp.0706396
  25. Ghobrial IM, Witzig TE, Adjei AA. Targeting apoptosis pathways in cancer therapy. CA: a cancer journal for clinicians. 2005;55(3):178-194. https://doi.org/10.3322/canjclin.55.3.178
  26. Guo W, et al. Targeting GRP75 improves HSP90 inhibitor efficacy by enhancing p53-mediated apoptosis in hepatocellular carcinoma. PloS one. 2014;9(1):e85766. https://doi.org/10.1371/journal.pone.0085766
  27. Ullah MF. Sulforaphane (SFN): an isothiocyanate in a cancer chemoprevention paradigm. Medicines. 2015;2(3):141-156. https://doi.org/10.3390/medicines2030141
  28. Qazi A, et al. Anticancer activity of a broccoli derivative, sulforaphane, in barrett adenocarcinoma: potential use in chemoprevention and as adjuvant in chemotherapy. Transl Oncol. 2010;3(6):389-99. https://doi.org/10.1593/tlo.10235
  29. Beyer-Sehlmeyer G, et al. Suppressive subtractive hybridisation reveals differential expression of serglycin, sorcin, bone marrow proteoglycan and prostate-tumour-inducing gene I (PTI-1) in drug-resistant and sensitive tumour cell lines of haematopoetic origin. Eur J Cancer. 1999;35(12):1735-42. https://doi.org/10.1016/S0959-8049(99)00202-6
  30. Bertram J, et al. Overexpression of ribosomal proteins L4 and L5 and the putative alternative elongation factor PTI-1 in the doxorubicin resistant human colon cancer cell line LoVoDxR. Eur J Cancer. 1998;34(5):731-6. https://doi.org/10.1016/S0959-8049(97)10081-8
  31. Zhang W, et al. Expressions of heat shock protein (HSP) family HSP 60, 70 and 90 a in colorectal cancer tissues and their correlations to pathohistological characteristics. Chinese J Cancer. 2009;28:1-7.
  32. Lim SO, et al. Expression of heat shock proteins (HSP27, HSP60, HSP70, HSP90, GRP78, GRP94) in hepatitis B virus-related hepatocellular carcinomas and dysplastic nodules. World journal of gastroenterology: WJG. 2005;11(14):2072-2079. https://doi.org/10.3748/wjg.v11.i14.2072
  33. Lebret T, et al. Heat shock proteins HSP27, HSP60, HSP70, and HSP90. Cancer. 2003;98(5):970-977. https://doi.org/10.1002/cncr.11594
  34. Yano M, et al. Expression and roles of heat shock proteins in human breast cancer. Cancer Science. 1996;87(9):908-915.
  35. Song CH, et al. Potential prognostic value of heat-shock protein 90 in the presence of phosphatidylinositol-3-kinase overexpression or loss of PTEN, in invasive breast cancers. Breast Cancer Research. 2010;12(2):R20. https://doi.org/10.1186/bcr2557
  36. Huang B, et al. Differential expression of estrogen receptor ${\alpha}$, ${\beta}1$, and ${\beta}2$ in lobular and ductal breast cancer. Proceedings of the National Academy of Sciences. 2014;111(5):1933-1938. https://doi.org/10.1073/pnas.1323719111
  37. Barrios C, et al. What is the role of chemotherapy in estrogen receptor-positive, advanced breast cancer? Annals of oncology. 2009;20(7):1157-1162. https://doi.org/10.1093/annonc/mdn756
  38. Taherian A, Mazoochi T. Different Expression of Extracellular Signal-Regulated Kinases (ERK) 1/2 and Phospho-Erk Proteins in MBA-MB-231 and MCF-7 Cells after Chemotherapy with Doxorubicin or Docetaxel. Iran J Basic Med Sci. 2012;15(1):669-77.
  39. Taherian A, Krone PH, Ovsenek N. A comparison of $Hsp90{\alpha}$ and $Hsp90{\beta}$ interactions with cochaperones and substrates. Biochemistry and Cell Biology. 2008;86(1):37-45. https://doi.org/10.1139/O07-154
  40. Didelot C, et al. Interaction of heat-shock protein $90{\beta}$ isoform ($HSP90{\beta}$) with cellular inhibitor of apoptosis 1 (c-IAP1) is required for cell differentiation. Cell Death Differ. 2008;15(5):859-66. https://doi.org/10.1038/cdd.2008.5
  41. Dong H, et al. Breast cancer MDA-MB-231 cells use secreted heat shock protein-90alpha ($Hsp90{\alpha}$) to survive a hostile hypoxic environment. Scientific reports. 2016;6.
  42. Romaniuk A, Lyndin M. Immune microenvironment as a factor of breast cancer progression. Diagnostic pathology. 2015;10(1):79. https://doi.org/10.1186/s13000-015-0316-y