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Effect of Cinobufotalin on Three Different Ovarian Cancer Cell Lines
Author(s) -
Afroze Syeda H,
Tobin Richard,
Peddaboina Chander,
McCormick Timothy C,
NewellRogers Martha K,
Zawieja David C,
Kuehl Thomas J,
Uddin Mohammad N
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.633.1
Subject(s) - viability assay , cell growth , cell culture , apoptosis , cell , cell sorting , microbiology and biotechnology , biology , ovarian cancer , mtt assay , cell cycle , cancer research , chemistry , cancer , flow cytometry , biochemistry , genetics
Objective Cinobufotalin (CINO), a cardiotonic steroid (CTS) or bufadienolide, is extracted from the skin secretions of the traditional Chinese medicine giant toads (Chan su). CINO has been used as a cardiotonic, diuretic and hemostatic agent. Previously we have shown that CINO inhibits cytotrophoblast cell function. Recently others have shown that CINO inhibits function of a lung cancer cell line. In this study, we assessed the effect of CINO on three different ovarian cancer cell lines; SK‐OV‐3, CRL‐1978 and CRL‐11731 to assess whether the effect of CINO is cell specific. Study Design We evaluated the effect of CINO on three ovarian cancer cells by treating cultures with 0.1, 1, 5, and 10 μM concentrations of CINO. Cell proliferation, migration and invasion were measured by using a CellTiter Assay (Promega), Cytoselect Assay (Cell Biolabs) and by using a FluoroBlock Assay (BD) respectively. Proliferating Cell Nuclear Antigen (PCNA) was evaluated in cell lysates of CINO treated cells by western blot analysis. Cell cycle arrest and cell viability were determined by fluorescence‐activated cell sorting (FACS) analysis. We performed Annexin V staining by immunofluorescence to evaluate the pro‐apoptotic protein expression. In addition mitochondrial membrane potential was measured using MMP kit and by FACS analysis. Results Concentrations of CINO at 0.5 μM inhibit SK‐OV‐3, CRL‐1978, and CRL‐11731 ovarian cancer cells proliferation, migration and invasion (p < 0.05) without cell death and loss of cell viability, but cell viability differed among cell lines. Each cell line differed in response to CINO doses for PCNA expression (p < 0.05) as well as Annexin V pro‐apoptotic protein expression. CINO decreases mitochondrial membrane potential (p < 0.05) for SK‐OV‐3. However, in CRL‐1978 and CRL‐11731, mitochondrial membrane potential (p < 0.05) is increased in response to CINO treatment. Conclusion CINO is cell specific, as each cancer cell line responds differently. These data demonstrate that the mode of action of CINO is different on these three types of ovarian cancer cells.