Comparative analysis of XTT assay and xCELLigence system by measuring cytotoxicity of resveratrol in human cancer cell lines

Harika Atmaca 1 , Emir Bozkurt 1 , Aslı Kısım 1  and Rüçhan Uslu 2
  • 1 Celal Bayar University, Faculty of Science and Letters, Department of Biology, Section of Molecular Biology, Manisa, Turkey
  • 2 Ege University, School of Medicine, Medical Oncology Department, Izmir, Turkey
Harika Atmaca, Emir Bozkurt, Aslı Kısım and Rüçhan Uslu

Abstract

Objective:

In vitro preliminary oncological and translational studies are mainly based on evaluating the cytotoxic effects of a specific compound on cultured cells. Resveratrol is a commercially available compound which is originally isolated from the roots of white hellebore and later from Polygonum cuspidatum. The objective of the study was to compare cytotoxicity data of Resveratrol from XTT end point assay with a real-time cell based xCELLigence system in terms of accuracy, sensitivity, speed and reproducibility in a panel of human cancer cell lines.

Methods:

XTT end point assay and real-time cell based xCELLigence system were used to evaluate cytotoxicity. Cytotoxicity results were verified by monitoring cells under phase-contrast microscope which were treated with IC50 values of resveratrol.

Results:

Resveratrol decreased cell viability in a time- and concentration-dependent manner in all cancer cell lines when tested by both the XTT assay and xCELLigence system. Standard deviations of the xCELLigence data were found to be lower than the data from XTT assay.

Conclusion:

The data from this study strongly imply that xCELLigence system has higher precision, more enlightening and more reproducible than XTT end point assay.

  • 1.

    Schröterová L, Králová V, Vorácová A, Hasková P, Rudolf E, Cervinka M. Antiproliferative effects of selenium compounds in colon cancer cells: comparison of different cytotoxicity assays. Toxicol In Vitro 2009;3:1406–11.

  • 2.

    Cory AH, Owen TC, Barltrop JA, Cory JG. Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture. Cancer Commun 1991;3:207–12.

    • Crossref
    • PubMed
    • Export Citation
  • 3.

    Urcan E, Haertel U, Styllouet M, Hickel R, Scherthan H, Reichl FX. Real-time xCELLigence impedance analysis of the cytotoxicity of dental composite components on human gingival fibroblasts. Dent Mater 2010;26:51–8.

    • Crossref
    • PubMed
    • Export Citation
  • 4.

    Jang M, Cai L, Udeani GO, Slowing KV, Thomas CF, Beecher CW, et al. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 1997;275:218–20.

    • Crossref
    • PubMed
    • Export Citation
  • 5.

    Howitz KT, Bitterman KJ, Cohen HY, Lamming DW, Lavu S, Wood JG, et al. Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan. Nature 2003;425:191–6.

    • Crossref
    • PubMed
    • Export Citation
  • 6.

    Wood JG, Rogina B, Lavu S, Howitz K, Helfand SL, Tatar M, et al. Sirtuin activators mimic caloric restriction and delay ageing in metazoans. Nature 2004;430:686–9.

    • Crossref
    • PubMed
    • Export Citation
  • 7.

    Li JP. Resveratrol caused apoptosis in QGY-7701 cells. Eur Rev Med Pharmacol Sci 2015;19:3303–8.

    • PubMed
    • Export Citation
  • 8.

    Khan A, Aljarbou AN, Aldebasi YH, Faisal SM, Khan MA. Resveratrol suppresses the proliferation of breast cancer cells by inhibiting fatty acid synthase signaling pathway. Cancer Epidemiol 2014;38:765–72.

    • Crossref
    • PubMed
    • Export Citation
  • 9.

    Roche Diagnostics GmbH. Introduction of the RTCA SP Instrument. RTCA SP Instrument Operator‘s Manual. A Acea Biosciences Inc. 2008;14–6.

  • 10.

    Xing JZ, Zhu L, Jackson JA, Gabos S, Sun XJ, Wang XB, et al. Dynamic monitoring on microelectronic sensors. Chem Res Toxicol 2005;18:154–61.

    • Crossref
    • PubMed
    • Export Citation
  • 11.

    Papadopoulos NG, Dedoussis GV, Spanakos G, Gritzapis AD, Baxevanis CN, Papamichail M. An improved fluorescence assay for the determination of lymphocyte-mediated cytotoxicity using flow cytometry. J Immunol Methods 1994;177:101–11.

    • Crossref
    • PubMed
    • Export Citation
  • 12.

    Stewart JR, O’Brian CA. Resveratrol antagonizes EGFR-dependent Erk1/2 activation in human androgen-independent prostate cancer cells with associated isozyme-selective PKC alpha inhibition. Invest New Drugs 2004;22:107–17.

    • Crossref
    • PubMed
    • Export Citation
  • 13.

    Rezk YA, Balulad SS, Keller RS, Bennett JA. Use of resveratrol to improve the effectiveness of cisplatin and doxorubicin: study in human gynecologic cancer cell lines and in rodent heart. Am J Obstet Gynecol 2006;194:23–6.

    • Crossref
    • Export Citation
  • 14.

    Yoo KM, Kim S, Moon BK, Kim S, Kim KT, Kim SY, et al. Potent inhibitory effects of resveratrol derivatives on progression of prostate cancer cells. Arch Pharm (Weinheim) 2006;339:238–41.

    • Crossref
    • PubMed
    • Export Citation
  • 15.

    He X, Wang Y, Zhu J, Orloff M, Eng C. Resveratrol enhances the anti-tumor activity of the mTOR inhibitor rapamycin in multiple breast cancer cell lines mainly by suppressing rapamycin-induced AKT signaling. Cancer Lett 2011;301:168–76.

    • Crossref
    • PubMed
    • Export Citation
Purchase article
Get instant unlimited access to the article.
$42.00
Log in
Already have access? Please log in.


or
Log in with your institution

Journal + Issues

Turkish Journal of Biochemistry (TJB), official journal of Turkish Biochemical Society, is issued electronically every 2 months. The main aim of the journal is to support the research and publishing culture by ensuring that every published manuscript has an added value and thus providing international acceptance of the “readability” of the manuscripts published in the journal.

Search