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Planta Med 2014; 80(08/09): 662-670
DOI: 10.1055/s-0034-1368506
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Human Ovarian Cancer Multicellular Spheroids: A Model for Testing Antiproliferation Activity of Devilʼs Club (Oplopanax horridus) and Anticancer Agents

Joseph Tai
1   Department of Pathology and Laboratory Medicine, Child and Family Research Institute, University of British Columbia, Canada
,
Susan S. C. Cheung
2   Department of Surgery, University of British Columbia, Canada
,
David Hasman
3   Forensic Science Center, British Columbia Institute of Technology, Burnaby, British Columbia, Canada
› Institutsangaben
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Publikationsverlauf

received 20. Dezember 2013
revised 17. April 2014

accepted 27. April 2014

Publikationsdatum:
12. Juni 2014 (online)

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Abstract

This study was conducted to employ an ovarian cancer Ovcar 10 three-dimensional model to assess the antiproliferation activity of the medicinal plant Devilʼs club, Oplopanax horridus, and its active compound, alone and in combination, with chemotherapeutic agents compared to Ovcar 10 two-dimensional cells grown as monolayer cells. Ovcar 10 three-dimensional spheroids were prepared with a rotary cell culture system. Cell counting kit-8 assessed the antiproliferation activity. Apoptosis-related gene expression in three-dimensional spheroids and two- dimensional cells was analyzed with an apoptosis antibody array. Flow cytometry was used to analyze the cell cycle. Ovcar 10 cells formed compact three-dimensional spheroids after 5 days of culture in a rotary culture system. Ovcar 10 three-dimensional spheroids were significantly more resistant to killing by Devilʼs club extract, its active compound alone, gemcitabine, and paclitaxel, but not cisplatin compared to two-dimensional cells, with IC50 levels closer to that observed in vivo. Devilʼs club extract and its active compound alone significantly enhanced the antiproliferation activity of cisplatin and gemcitabine at some concentrations, but did not affect the activity of paclitaxel. A number of apoptosis-related genes were differentially expressed in three-dimensional spheroids, two-dimensional cells, and cells treated with Devilʼs club extract compared to untreated controls. In three-dimensional spheroids, the proportion of cells in the G2/M phase was slightly increased and the S phase was slightly decreased compared to two-dimensional cells. Ovcar 10 cells in three-dimensional spheroids altered the expression of multiple apoptosis-related genes, which may have contributed to the increased resistance of the cells to some drugs.

Key words

Araliaceae - Oplopanax horridus - human ovarian cancer three-dimensional spheroids - North American herbal medicine - antiproliferation - anticancer agents
 

  • References

  • 1 United States Centers for Disease Control and Prevention. Available at http://www.cdc.gov/cancer/ovarian/statistics/ . Accessed May 17, 2014.
    PubMed
  • 2 Loessner D, Stok KS, Lutolf MP, Hutmacher DW, Clements JA, Rizzi SC. Bioengineered 3D platform to explore cell-ECM interactions and drug resistance of epithelial ovarian cancer cells. Biomaterials 2010; 31: 8494-8506
    CrossrefPubMedSuche in Google Scholar
  • 3 Schmeichel KL, Bissell MJ. Modeling tissue-specific signalling and organ function in three dimensions. J Cell Sci 2003; 116: 2377-2388
    CrossrefPubMedSuche in Google Scholar
  • 4 Nyga A, Cheema U, Loizidou M. 3D tumour models: novel in vitro approaches to cancer studies. J Cell Commun Signal 2011; 5: 239-248
    CrossrefPubMedSuche in Google Scholar
  • 5 Lantz TC, Swerhun K, Turner NJ. Devilsʼs Club (Oplopanax horridus): An Ethnobotanical Review. HerbalGram 2004; 62: 33-48
    PubMedSuche in Google Scholar
  • 6 Tai J, Cheung S, Cheah S, Chan E, Hasman D. In vitro anti-proliferative and antioxidant studies on Devilʼs Club Oplopanax horridus . J Ethnopharmacol 2006; 108: 228-235
    CrossrefPubMedSuche in Google Scholar
  • 7 Tai J, Cheung S, Chan E, Hasman D. Inhibition of human ovarian cancer cell lines by Devilʼs club Oplopanax horridus . J Ethnopharmacol 2010; 127: 478-485
    CrossrefPubMedSuche in Google Scholar
  • 8 Sun S, Du GJ, Qi LW, Williams S, Wang CZ, Yuan CS. Hydrophobic constituents and their potential anticancer activities from Devilʼs club (Oplopanax horridus Miq.). J Ethnopharmacol 2010; 132: 280-285
    CrossrefPubMedSuche in Google Scholar
  • 9 Reid JM, Qu W, Safgren SL, Ames MM, Krailo MK, Seibel NL, Kuttesch J, Holcenberg J. Phase I trial and pharmacokinetics of gemcitabine in children with advanced solid tumors. J Clin Oncol 2004; 22: 2445-2451
    CrossrefPubMedSuche in Google Scholar
  • 10 Monjanel-Mouterde S, Ciccolini J, Bagarry D, Zonta-David M, Duffaud F, Favre R, Durand A. Population pharmacokinetics of cisplatin after 120-h infusion: application to routine adaptive control with feedback. J Clin Pharm Therap 2003; 28: 109-116
    CrossrefPubMedSuche in Google Scholar
  • 11 Ichiki M, Gohara R, Fujiki R, Hoashi S, Rikimaru T, Aizawa H. Phase I and pharmacokinetic study of carboplatin and paclitaxel with a biweekly schedule in patients with advanced non-small-cell lung cancer. Cancer Chemother Pharmacol 2003; 52: 67-72
    CrossrefPubMedSuche in Google Scholar
  • 12 Elliott NT, Yuan F. A review of three-dimensional in vitro tissue models for drug discovery and transport studies. J Pharm Sci 2011; 100: 59-74
    CrossrefPubMedSuche in Google Scholar
  • 13 Nirmalanandhan VS, Duren A, Hendricks P, Vielhauer G, Sittampalam GS. Activity of anticancer agents in a three-dimensional cell culture model. Assay Drug Dev Tech 2010; 8: 581-590
    CrossrefPubMedSuche in Google Scholar
  • 14 Kumar HR, Zhong X, Hoelz DJ, Rescorla FJ, Hickey RJ, Malkas LH, Sandoval JA. Three-dimensional neuroblastoma cell culture: proteomic analysis between monolayer and multicellular tumor spheroids. Pediatr Surg Int 2008; 24: 1229-1234
    CrossrefPubMedSuche in Google Scholar
  • 15 Yamada KM, Cukierman E. Modeling tissue morphogenesis and cancer in 3D. Cell 2007; 24: 601-610
    PubMedSuche in Google Scholar
  • 16 Sutherland RM. Cell and environment interactions in tumor microregions: the multicell spheroid model. Science 1988; 240: 177-184
    CrossrefPubMedSuche in Google Scholar
  • 17 Ingram M, Techy GB, Saroufeem R, Yazan O, Narayan KS, Goodwin TJ, Spaulding GF. Three-dimensional growth patterns of human tumor cell line in simulated microgravity of a NASA bioreactor. In Vitro Cell Biol Anim 1997; 33: 459-466
    PubMedSuche in Google Scholar
  • 18 Kunz-Schughart LA, Freyer JP, Hofstaedter F, Ebner R. The use of 3-D cultures for high-throughput screening: the multicellular spheroid model. J Biomol Screen 2004; 9: 273-285
    CrossrefPubMedSuche in Google Scholar
  • 19 Timmins NE, Dietmair S, Nielsen LK. Hanging-drop multicellular spheroids as a model of tumour angiogenesis. Angiogenesis 2004; 7: 97-103
    CrossrefPubMedSuche in Google Scholar
  • 20 Kobaisy M, Abramowski Z, Lermer L, Saxena G, Hancock RE, Towers GH, Doxsee D, Stokes RW. Antimycobacterial polyynes of Devilʼs club (Oplopanax horridus), a North American native medicinal plant. J Nat Prod 1997; 60: 1210-1213
    CrossrefPubMedSuche in Google Scholar
  • 21 Tai J, Cheung SS, Ou D, Warnock GL, Hasman D. Antiproliferation activity of Devilʼs club (Oplopanax horridus) and anticancer agents on human pancreatic cancer multicellular spheroids. Phytomedicine 2014; 21: 506-514
    CrossrefPubMedSuche in Google Scholar
  • 22 Wang CZ, Zhang Z, Huang WH, Du DJ, Wen XD, Calway T, Yu C, Nass R, Zhao J, Du W, Li SP, Yuan CS. Identification of potential anticancer compounds from Oplopanax horridus . Phytomedicine 2013; 20: 999-1006
    CrossrefPubMedSuche in Google Scholar
  • 23 Yang Z, Zhao X. A 3D model of ovarian cancer cell lines on peptide nanofiber scaffold to explore the cell-scaffold interaction and chemotherapeutic resistance of anticancer drugs. Int J Nanomedicine 2011; 6: 303-310
    CrossrefPubMedSuche in Google Scholar
  • 24 Horning JL, Sahoo SK, Vijayaraghavalu S, Dimitrijevic S, Vasir JK, Jain TK, Pand AK, Labhasetwar V. 3-D tumor model for in vitro evaluation of anticancer drugs. Mol Pharm 2008; 5: 849-862
    CrossrefPubMedSuche in Google Scholar
  • 25 LaRue KEA, Khalil M, Freyer JP. Microenvironmental regulation of proliferation in multicellular spheroids is mediated through differential expression of cyclin-dependent kinase inhibitors. Cancer Res 2004; 64: 1621-1631
    CrossrefPubMedSuche in Google Scholar
  • 26 Walker J, Martin C, Callaghan R. Inhibition of P-glycoprotein function by XR9576 in a solid tumour model can restore anticancer drug efficacy. Eur J Cancer 2004; 40: 594-605
    CrossrefPubMedSuche in Google Scholar
  • 27 Smalley KS, Lioni M, Herlyn M. Life isnʼt flat: taking cancer biology to the next dimension. In Vitro Cell Dev Biol Anim 2006; 42: 242-247
    CrossrefPubMedSuche in Google Scholar
  • 28 Nelson CM, Bissel MJ. Modeling dynamic reciprocity: engineering three-dimensional culture models of breast architecture, function, and neoplastic transformation. Semin Cancer Biol 2005; 15: 342-352
    CrossrefPubMedSuche in Google Scholar
  • 29 Zietarska M, Maugard CM, Filali-Mouhim A, Alam-Fahmy M, Tonin PN, Provencher DM, Mes-Masson AM. Molecular description of a 3D in vitro model for the study of epithelial ovarian cancer (EOC). Mol Carcinogenesis 2007; 46: 872-885
    CrossrefPubMedSuche in Google Scholar
  • 30 Zurawa-Janicka D, Skorko-Glonek J, Lipinska B. HtrA proteins as targets in therapy of cancer and other diseases. Expert Opin Ther Targets 2010; 14: 665-679
    CrossrefPubMedSuche in Google Scholar
  • 31 Kongpetch S, Kukongviriyapen V, Prawan A, Senggunprai L, Kukongviriyapan U, Buranrat B. Crucial role of heme oxygenase-1 on the sensitivity of cholangiocarcinoma cells to chemotherapeutic agents. PLoS One 2012; 7: e34994
    CrossrefPubMedSuche in Google Scholar
  • 32 Hassan MK, Watari H, Han Y, Mitamura T, Hosaka M, Wang L, Tanaka S, Sakuragi N. Clusterin is a potential molecular predictor for ovarian cancer patientʼs survival: targeting clusterin improves response to paclitaxel. J Exp Clin Cancer Res 2011; 30: 113
    CrossrefPubMedSuche in Google Scholar
  • 33 Glorieux C, Dejeans N, Sid B, Beck R, Calderon PB, Verrax J. Catalase overexpression in mammary cancer cells leads to a less aggressive phenotype and an altered response to chemotherapy. Biochem Pharmacol 2011; 82: 1384-1390
    CrossrefPubMedSuche in Google Scholar
  • 34 Yang D, Torres CM, Bardhan K, Zimmerman M, McGaha TL, Liu K. Decitabine and vorinostat cooperate to sensitize colon carcinoma cells to Fas ligand-induced apoptosis in vitro and tumor suppression in vivo . J Immunol 2012; 188: 4441-4449
    CrossrefPubMedSuche in Google Scholar
  • 35 Cui J, Nan KJ, Tian T, Guo YH, Zhao N, Wang L. Chinese medicinal compound delisheng has satisfactory anti-tumor activity, and is associated with up-regulation of endostatin in human hepatocellular carcinoma cell line HepG2 in three-dimensional culture. World J Gastroenterol 2007; 13: 5432-5439
    CrossrefPubMedSuche in Google Scholar
  • 36 Mueller-Klieser W, Schreiber-Klais S, Walenta S, Kreuter MH. Bioactivity of well-defined green tea extracts in multicellular tumor spheroids. Int J Oncol 2002; 21: 1307-1315
    PubMedSuche in Google Scholar
  • 37 Kueng W, Silber E, Eppenberger U. Quantification of cells cultured on 96-well plates. Anal Biochem 1989; 182: 16-19
    CrossrefPubMedSuche in Google Scholar
  • 38 Berenbaum MC. What is synergy?. Pharmacol Rev 1989; 41: 93-141
    PubMedSuche in Google Scholar
  • 39 Wagner H, Ulrich-Merzenich G. Synergy research: approaching a new generation of phytochemicals. Phytomedicine 2009; 16: 97-110
    CrossrefPubMedSuche in Google Scholar