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CINOBUFOTALIN INHIBITS OVARIAN CANCER CELL METASTASIS VIA APOPTOTIC SIGNALING AND TARGETING THE mTOR PATHWAY
Author(s) -
Afroze Syeda Humayra,
Peddaboina Chander,
Dean DeShaundra M,
McDowell Anthony,
McCormick Timothy C,
NewellRogers Karen,
Zawieja David C,
Kuehl Thomas J,
Uddin Mohammad N
Publication year - 2017
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.31.1_supplement.934.20
Subject(s) - cell growth , ovarian cancer , viability assay , apoptosis , pi3k/akt/mtor pathway , cancer research , cell culture , cell , biology , mtt assay , cancer cell , cell cycle , chemistry , microbiology and biotechnology , cancer , 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 a hemostatic agent. Previously we have shown that CINO inhibits the cytotrophoblast cell function. Recently other study has shown that CINO inhibits A549, a lung cancer cell function. We demonstrated that CINO inhibited the proliferation, migration and invasion of three ovarian cancer cell lines; SK‐OV‐3, CRL‐1978 and CRL‐11731. In this study we evaluated the cellular mechanisms by which CINO inhibits ovarian cancer cell function. Study Design We evaluated the effect of CINO on three ovarian cancer cells SK‐OV‐3, CRL‐1978, and CRL‐11731 function in vitro. Each Cell lines were treated with different concentrations of CINO (0.1, 1, 5 and 10 μM). For each cell line 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), mTOR and pAKT were also evaluated in cell lysates of CINO treated these 3 ovarian cancer cells by western blot analysis. Cell Cycle arrest and Cell viability were determined by fluorescence‐activated cell sorting (FACS) analysis. We also performed Annexin V staining on CINO treated these 3 ovarian cancer cell lines by immunofluorescence to evaluate the pro‐apoptotic protein expression. In addition mitochondrial membrane potential (MMP) has also been measured for all these 3 ovarian cell lines after CINO treatment using MMP kit, by FACS analysis. Results Concentration of CINO at 0.5μM inhibit SK‐OV‐3, CRL‐1978, and CRL‐11731 ovarian cancer cells proliferation, migration and invasion without cell death and loss of cell viability but cell viability differs for each cell line. Each cell lines differ in response to CINO doses for PCNA expression as well as Annexin V pro‐apoptotic protein expression. CINO decreases mitochondrial membrane potential for SK‐OV‐3 but not for CRL‐1978 and CRL‐11731 increases in response to CINO treatment. CINO causes the decrease in mTOR and pAKT expression in SK‐OV‐3 and CRL‐1978 but not in CRL‐11731. 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 3 types of ovarian cancer cells. Moreover, it also can be suggested that CINO inhibits the ovarian cancer cell metastasis via apoptotic signaling and targeting the mTOR pathway. Support or Funding Information The Discovery Foundation Grant (MNU), Grimes Foundation Endowment Award (MNU)