EF24’ün Dosetaksel ile Sıralı Uygulanmasının Metastatik Meme Kanser Hücre Hattına Etkisi

Amaç: Taksan sınıfı antineoplastik bir ajan olan dosetaksel, son yılların en önemli kemoterapötik ajanlarından biridir. Günümüzde, meme kanseri tedavisi için klasik kemoterapi ajanlarının yeni moleküller ile birlikte kullanılması sonucu daha etkili tedavi seçenekleri belirlenmeye çalışılmaktadır. Bu moleküllerin bir tanesi de sentetik kurkumin analoğu olan EF24’tür. Çalışmamızda, dosetaksel’in tek başına ve EF24 ön koşullamasının ardından dosetaksel uygulamasının, meme kanseri MCF-7 hücrelerindeki hücre çoğalması ve apoptoz üzerindeki etkilerinin belirlenmesi amaçlanmıştır. Gereç ve Yöntem: EF24 ve dosetaksel’in hücre canlılığı ve sitotoksik etki/etkilerini belirlemek amacıyla sırasıyla 3-(4,5-dimetiltiazol-2-yl)-2,5-difeniltettrazoliyum bromür (MTT) ve laktat dehidrojenaz (LDH) testleri uygulandı. Hücre canlılığı ve sitotoksisite testinin sonucunda seçilen uygun dozlar hücrelere uygulandıktan sonra akım sitometri yöntemi ile hücre ölüm analizi yapıldı. Bunun yanı sıra kantitatif gerçek zamanlı polimeraz zincir reaksiyonu analizi (qRT-PCR) ile cMYC ve cFOS genlerinin mRNA düzeyindeki ifadeleri belirlendi. Bulgular: MCF-7 hücrelerine EF24 ön uygulamasının ardından dosetaksel uygulamasının, dosetakselin tek başına uygulanmasına kıyasla hücre canlılığını anlamlı ölçüde düşürdüğü belirlendi. Apoptotik ve nekrotik hücre oranları akım sitometri yöntemi ile belirlendi. Buna ilaveten, EF24 ön koşullamasının, tek başına dosetaksel uygulamasına kıyasla cMYC ve cFOS genlerinin mRNA düzeylerini anlamlı derecede azalttığı belirlendi. Sonuç: MCF-7 hücrelerine EF24 ön uygulamasının, dosetaksel’in tek başına uygulamasına kıyasla hücreyi daha duyarlı hale getirmesi bu uygulama-nın meme kanser tedavisinde kullanılabileceğini düşündürmektedir. Ancak ilacın meme kanser tedavisinde daha etkin kullanımı için deneysel in vitro ve in vivo modellere gereksinim duyulmaktadır.

The Effects of Sequential Administration of EF24 with Docetaxel Apoptotic Response in Metastatic Breast Cancer Cell Line

Objective: Docetaxel, a taxane class agent, has become one of the most important chemotherapeutic agents in the past several years. Currently, more effective treatment options have been investigated by combining new molecules with the classic chemotherapy agents for breast cancer. One of these molecules is EF24, which is a synthetic curcumin analog. We aimed to investigate possible antiproliferative and apoptotic effects of EF24 and docet-axel alone as well as with their sequential administration on MCF-7 breast cancer cells. Material and Method: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Lactate dehydrogenase (LDH) tests were per-formed to determine the effect of EF24 and docetaxel on cell viability and cytotoxicity. Apoptotic cell death methods were performed using Flow cytometry assay, with selected appropriate doses of the cells. Besides, relative mRNA levels of cMYC and cFOS genes were determined by quantita-tive Real-time PCR method (qRT-PCR). Results: The viability of MCF-7 cells decreased significantly after treatment with sequential administration of docetaxel followed by EF24 treat-ments, compared to docetaxel alone. Compared to docetaxel alone treatment, EF24 pretreatment overwhelmingly increased apoptosis level in MCF-7 cells. The percentage of apoptotic and necrotic cells were determined by the flow cytometer analysis. Moreover, real-time PCR analyses showed that cMYC and cFOS mRNA levels changed markedly after sequential treatment. Conclusion: These data suggest that sequential administration of EF24 with docetaxel could be useful as a potential chemotherapeutic agents in the management of breast cancer. Further analyses using in vitro and in vivo models are needed to confirm these findings.

___

  • 1. Zhao X, Gurumurthy CB, Malhotra G, et al. Breast cancer subtypes: two decades of journey from cell culture to patients. Adv Exp Med Biol 2011; 720: 135-44.
  • 2. Aziz MY, Abu N, Yeap SK, et al. Combinato-rial Cytotoxic Effects of Damnacanthal and Doxorubicin against Human Breast Cancer MCF-7 Cells in Vitro. Molecules 2016; 21: 1-15.
  • 3. Xie ZZ, Li MM, Deng PF, et al. Paris saponin-induced autophagy promotes breast cancer cell apoptosis via the Akt/mTOR signaling pathway. Chem Biol Interact 2017; 12: 1-9.
  • 4. Salih AK, Fentiman IS. Breast cancer preven-tion: present and future. Cancer Threatment Rewievs 2001; 27: 261-73.
  • 5. Somunoğlu S. Meme kanseri: belirtileri ve er-ken tanıda kullanılan tarama yöntemleri. Fırat Sağlık Hizmetleri Dergisi 2009; 4: 103-22.
  • 6. Sasco AJ. Epidemiology of breast cancer: an environmental disease? Epidemiology of breast cancer: an environmental disease? APMIS 2001; 109: 80-92.
  • 7. Krishan A, Fitz CM, Andritsch I. Drug reten-tion, efflux, and resistance in tumor cells. Cy-tometry 1997; 29: 279-85.
  • 8. Baguley BC. Classical and targeted anticancer drugs: an appraisal of mechanisms of multidrug resistance. Methods Mol Biol 2016; 1395: 19-37.
  • 9. Fruci D, Cho WC, Nobili V, Locatelli F, Alisi A. Drug transporters and multiple drug resistan-ce in pediatric solid tumors. Curr Drug Metab 2016; 17: 308-16.
  • 10. Wu C, Gong MQ, Liu BY, Zhuo RX, Cheng SX. Co-delivery of multiple drug resistance in-hibitors by polymer/inorganic hybrid nanopar-ticles to effectively reverse cancer drug resistan-ce. Colloids Surf B Biointerfaces 2017; 149: 250-9.
  • 11. Strobl JS, Wonderlin WF, Flynn DC. Mitogenic signal transduction in human breast cancer cells. Gen Pharmacol 1995; 26: 1643-9.
  • 12. Torry DS, Cooper GM. Proto-oncogenes in development and cancer. Am J Reprod Immunol 1991; 25: 129-32.
  • 13. Anderson MW, Reynolds SH, You M, Maron-pot RM. Role of proto-oncogene activation in carcinogenesis. Environ Health Perspect 1992; 98: 13-24.
  • 14. Nestal de Moraes G, Delbue D, Silva KL, et al. FOXM1 targets XIAP and Survivin to modulate breast cancer survival and chemoresistance. Cell Signal 2015; 27: 2496-505.
  • 15. Maeda S, Saimura M, Minami S, et al. Efficacy and safety of eribulin as first- to third-line tre-atment in patients with advanced or metastatic breast cancer previously treated with anthracyc-lines and taxanes. Breast 2017; 32: 66-72.
  • 16. Zhang T, Wang R, Liu Y, Huang J, Yang Z. Efficacy and safety of doublet versus single agent as salvage treatment for metastatic breast cancer pretreated with anthracyclines and taxa-nes: a systematic review and meta-analysis. Curr Med Res Opin 2016; 32: 1883-9.
  • 17. Rincón R, Zazo S, Chamizo C, et al. c-Jun N-Terminal Kinase inactivation by Mitogen-Activated Protein Kinase Phosphatase 1 deter-mines resistance to Taxanes and Anthracyclines in breast cancer. Mol Cancer Ther 2016; 15: 2780-90.
  • 18. De Iuliis F, Salerno G, Giuffrida A, et al. Breast cancer cells respond differently to docetaxel de-pending on their phenotype and on survivin up-regulation. Tumour Biol 2016; 37: 2603-11.
  • 19. Morse DL, Gray H, Payne CM, Gillies RJ. Do-cetaxel induces cell death through mitotic ca-tastrophe in human breast cancer cells. Mol Cancer Ther 2005; 4: 1495-504.
  • 20. Lyseng-Williamson KA, Fenton C. Docetaxel: a review of its use in metastatic breast cancer. Drugs 2005; 65: 2513-31.
  • 21. Karampeazis A, Vamvakas L, Agelidou A, et al. Docetaxel vs. vinorelbine in elderly patients with advanced nonsmall-cell lung cancer: A Hellenic Oncology Research Group Randomi-zed Phase III Study. Clin Lung Cancer 2011; 12: 155-60.
  • 22. Yang F, Luo LJ, Zhang L, et al. MiR-346 pro-motes the biological function of breast cancer cells by targeting SRCIN1 and reduces chemo-sensitivity to docetaxel. Gene 2017; 600: 21-8.
  • 23. Zhang Y, Wang Y, Wei Y, et al. MiR-129-3p promotes docetaxel resistance of breast cancer cells via CP110 inhibition. Sci Rep 2015; 5: 15424.
  • 24. Wang X, Xu C, Hua Y, et al. Exosomes play an important role in the process of psoralen reverse multidrug resistance of breast cancer. J Exp Clin Cancer Res 2016; 35: 186.
  • 25. Hansen SN, Westergaard D, Thomsen MB, et al. Acquisition of docetaxel resistance in breast cancer cells reveals upregulation of ABCB1 expression as a key mediator of resistance ac-companied by discrete upregulation of other specific genes and pathways. Tumour Biol 2015; 36: 4327-38.
  • 26. Zhao P, Ma W, Hu Z, Zang L, Tian Z, Zhang K. Filamin A (FLNA) modulates chemosensitivity to docetaxel in triple-negative breast cancer through the MAPK/ERK pathway. Tumour Biol 2016; 37: 5107-15.
  • 27. Su S, Ding Y, Li Y, Wu Y, Nie G. Integration of photothermal therapy and synergistic che-motherapy by a porphyrin self-assembled micel-le confers chemosensitivity in triple-negative breast cancer. Biomaterials 2016; 80: 169-78.
  • 28. Singel SM, Cornelius C, Batten K, et al. A tar-geted RNAi screen of the breast cancer genome identifies KIF14 and TLN1 as genes that modu-late docetaxel chemosensitivity in triple-negative breast cancer. Clin Cancer Res 2013; 19: 2061-70.
  • 29. Selvendiran K, Tong L, Vishwanath S, et al. EF24 induces G2/M arrest and apoptosis in cisplatin-resistant human ovarian cancer cells by increasing PTEN expression. J Biol Chem 2007; 282: 28609–18.
  • 30. Tan X, Sidell N, Mancini A, et al. Multiple anticancer activities of EF24, a novel curcumin analog, on human ovarian carcinoma cells. Rep-rod Sci 2010; 17: 931-40.
  • 31. Zhang D, Wang Y, Dong L, et al. Therapeutic role of EF24 targeting glucose transporter 1-mediated metabolism and metastasis in ovarian cancer cells. Canc Sci 2013; 104: 1690-6.
  • 32. Subramaniam D, May R, Sureban SM, et al. Diphenyl difluoroketone: a curcumin derivative with potent in vivo anticancer activity. Canc Res 2008; 68: 1962-9.
  • 33. Yang CH, Yue J, Sims M, Pfeffer LM. The curcumin analog EF24 targets NF-B and miR-NA-21, and has potent anticancer activity in vit-ro and in vivo. PLoS One 2013; 8: e71130.
  • 34. Sun A, Lu YJ, Hu H, Shoji M, Liotta DC, Snyder JP. Curcumin analog cytotoxicity aga-inst breast cancer cells: exploitation of a redox-dependent mechanism. Bioorg Med Chem Lett 2009; 19: 6627-31.
  • 35. Yar Saglam AS, Yilmaz A, Onen HI, Alp E, Kayhan H, Ekmekci A. HDAC inhibitors, MS-275 and salermide, potentiates the anticancer ef-fect of EF24 in human pancreatic cancer cells. EXCLI J 2016; 15: 246-55.
  • 36. Onen HI, Yilmaz A, Alp E, et al. EF24 and RAD001 potentiates the anticancer effect of pla-tinum-based agents in human malignant pleural mesothelioma (MSTO-211H) cells and protects nonmalignant mesothelial (MET-5A) cells. Hum Exp Toxicol 2015; 34: 117-26.
  • 37. Thomas SL, Zhao J, Li Z, et al. Activation of the p38 pathway by a novel monoketone curcu-min analog, EF24, suggests a potential combi-nation strategy. Biochem Pharmacol 2010; 80: 1309-16.
  • 38. van Meerloo J, Kaspers GJ, Cloos J. Cell sensi-tivity assays: the MTT assay. Methods Mol Biol 2011; 731: 237-45.
  • 39. Smith SM, Wunder MB, Norris DA, Shellman YG. A simple protocol for using a LDH-based cytotoxicity assay to assess the effects of death and growth inhibition at the same time. PLoS One 2011; 6: e26908.
  • 40. Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 2002; 30: e36.
  • 41. Adams B, Herold M, Ferstl E, et al. Anticancer effects of monocarbonyl analogs of curcumin: oxidative stress, nuclear translocation and mo-dulation of AP-1 and NF-κB. Int J Cancer Ther Oncol 2015; 3: 1-15.
  • 42. Yang SJ, Lee SA, Park MG, et al. Induction of apoptosis by diphenyldifluoroketone in osteo-genic sarcoma cells is associated with activation of caspases. Oncol Rep 2014; 31: 2286-92.
  • 43. Lee CC, Houghton P. Cytotoxicity of plants from Malaysia and Thailand used traditionally to treat cancer. J Ethnopharmacol 2005; 100: 237-43.
  • 44. Nathwani SM, Butler S, Fayne D, et al. Novel microtubule-targeting agents, pyrrolo-1,5-benzoxazepines, induce apoptosis in multi-drug-resistant cancer cells. Cancer Chemother Phar-macol 2010; 66: 585-96.
  • 45. Vuorelaa P, Leinonenb M, Saikkuc P, et al. Natural products in the process of finding new drug candidates. Curr Med Chem 2004; 11: 1375-89.
  • 46. Fury MG, Pfister DG. Current recommendations for systemic therapy of recurrent and/or metas-tatic head and neck squamous cell cancer. J Natl Compr Canc Netw 2011; 9: 681-9.
  • 47. Fabbri F, Amadori D, Carloni S, et al. Mitotic catastrophe and apoptosis induced by docetaxel in hormone-refractory prostate cancer cells. J Cell Physiol 2008; 217: 494-501.
  • 48. Grimer R, Judson I, Peake D, Seddon B. Guide-lines for the management of soft tissue sarco-mas. Sarcoma 2010; 2010: 5061-82.
  • 49. Morse DL, Gray H, Payne CM, Gillies RJ. Do-cetaxel induces cell death through mitotic ca-tastrophe in human breast cancer cells. Mol Cancer Ther 2005; 4: 1495-504.
  • 50. Yeung TK, Germond C, Chen X, Wang Z. The mode of action of taxol: apoptosis at low con-centration and necrosis at high concentration. Biochem Biophys Res Commun 1999; 263: 398-404.
  • 51. Wang H, Vo T, Hajar A, et al. Multiple mecha-nisms underlying acquired resistance to taxanes in selected docetaxel-resistant MCF-7 breast cancer cells. BMC Cancer 2014; 22: 14-37.
  • 52. Hernández-Vargas H, Palacios J, Moreno-Bueno G. Molecular profiling of docetaxel cyto-toxicity in breast cancer cells: uncoupling of aberrant mitosis and apoptosis. Oncogene 2007; 26: 2902-13.
  • 53. Trebunova M, Laputkova G, Slaba E, Lacjakova K, Verebova A. Effects of docetaxel, doxorubi-cin and cyclophosphamide on human breast cancer cell line MCF-7. Anticancer Res 2012; 32: 2849-54.
  • 54. Chen W, Zou P, Zhao Z, et al. Synergistic anti-tumor activity of rapamycin and EF24 via incre-asing ROS for the treatment of gastric cancer. Redox Biol 2016; 10: 78-89.
  • 55. Liang Y, Yin D, Hou L, et al. Diphenyl difluo-roketone: a potent chemotherapy candidate for human hepatocellular carcinoma. PLoS One 2011; 6: e23908.
  • 56. Adams BK, Cai J, Armstrong J, et al. EF24, a novel synthetic curcumin analog, induces apop-tosis in cancer cells via a redox-dependent mec-hanism. Anticancer Drugs 2005; 16: 263-75.
  • 57. Chen X, Dai X, Zou P, et al. Curcuminoid EF24 enhances the anti-tumour activity of Akt inhibi-tor MK-2206 through ROS-mediated endoplas-mic reticulum stress and mitochondrial dysfunc-tion in gastric cancer. Br J Pharmacol 2017 Mar 3 [Epub ahead of print].
  • 58. Jochum W, Passegué E, Wagner EF. AP-1 in mouse development and tumorigenesis. Onco-gene 2001; 20: 2401-12. 59. Shaulian E, Karin M. AP-1 in cell proliferation and survival. Oncogene 2001; 20: 2390-400.
  • 60. De Sousa SO, Mesquita RA, Pinto DS Jr, Gut-kind S. Immunolocalization of c-Fos and c-Jun in human oral mucosa and in oral squamous cell carcinoma. J Oral Pathol Med 2002; 31: 78-81.
  • 61. el Khyari S, Bourgarel V, Barra Y, Braguer D, Briand C. Pretreatment by tubulin agents decre-ases C-MYC induction in human colon carci-noma cell line HT29-D4. Biochem Biophys Res Commun 1997; 231: 751-4.
  • 62. Li Y, Shi T, Zhao W. The mechanism of do-cetaxel-induced apoptosis in human lung cancer cells. Zhonghua Zhong Liu Za Zhi 2000; 22: 208-11.
  • 63. Kim SM, Lee SY, Yuk DY, et al. Inhibition of NF-kappaB by ginsenoside Rg3 enhances the susceptibility of colon cancer cells to docetaxel. Arch Pharm Res 2009; 32: 755-65.
  • 64. “Docetaxel With or Without a Phytochemical in Treating Patients With Breast Cancer”. https://clinicaltrials.gov/ct2/show/NCT00852332/ 18.04.2017.
  • 65. “Curcumin" in Combination With Chemothe-rapy in Advanced Breast Cancer”. https://clinicaltrials.gov/ct2/show/NCT03072992/ 18.04.2017.
Fırat Tıp Dergisi-Cover
  • ISSN: 1300-9818
  • Başlangıç: 2015
  • Yayıncı: Fırat Üniversitesi Tıp Fakültesi
Sayıdaki Diğer Makaleler

Dactylorhiza osmanica’nın Topraküstü Kısımlarında Antioksidan, Antimikrobiyal ve Tirozinaz İnhibitör Aktivitelerinin Araştırılması

REZZAN ALİYAZICIOĞLU, Nuriye KORKMAZ, Şeyda AKKAYA, SILA ÖZLEM ŞENER, UFUK ÖZGEN, ŞENGÜL ALPAY KARAOĞLU

Elazığ ve Çevresinde Avcılarda Tularemi Görülme Sıklığının Mikroaglütinasyon Yöntemiyle Araştırılması

Zülfü BAYAR, Mehmet AKSU, MUSTAFA YILMAZ

Tanısal Karmaşıklığa Sebep Olan Peritoneal Diyalize Bağlı Peritonit ve Pnömatozis İntestinalis Birlikteliği

Halil İbrahim TAŞCI

İskemik İnme Hastalarında Ekstrakraniyal Karotis Arter Stentleme: Tek Merkez Deneyimi

EMRAH AYTAÇ, Çetin Kürşat AKPINAR, MURAT GÖNEN

Üniversite Öğrencilerinin Nargile İçme Konusundaki Bilgi, Tutum ve Davranışları

Ayhan AKTAŞ, SEYHAN HIDIROĞLU, MELDA KARAVUŞ

Fibrokistik Değişiklikler Masum mudur? Memenin Fibrokistik Değişikliklerinde HER-2’nin Önemi ve Meme Kanseri ile İlişkisi

Özgen ARSLAN SOLMAZ

Edinsel Anatomik Bozukluğa Bağlı Çekal Volvulus Olgusu

Talha SARİGÖZ, Ramazan AZAR, Yusuf SEVİM, İnanç Şamil SARICI, TAMER ERTAN, ÖMER TOPUZ

Mide Kanseri Tedavisinde Açık ve Laparoskopik Distal Subtotal Gastrektominin Kısa Dönem Onkolojik Sonuçları

İLHAN ECE, Fahrettin ACAR, HÜSEYİN YILMAZ, BAYRAM ÇOLAK, SERDAR YORMAZ, MUSTAFA ŞAHİN

Are Fibrocystic Changes Innocent? Importance of HER-2 in Fibrocystic Changes of Breast and Correlation Breast Cancer Risk

ÖZGEN ARSLAN SOLMAZ

Süt Çocukluğu Döneminde Akrep Sokması: Olgu Sunumu

Mehmet Yusuf SARI, MEHMET KILIÇ, Mustafa AYDIN, ERDAL TAŞKIN