Paclitaxel and the dietary flavonoid fisetin: a synergistic combination that induces mitotic catastrophe and autophagic cell death in A549 non-small cell lung cancer cellsReportar como inadecuado

Paclitaxel and the dietary flavonoid fisetin: a synergistic combination that induces mitotic catastrophe and autophagic cell death in A549 non-small cell lung cancer cells - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Cancer Cell International

, 16:10

First Online: 16 February 2016Received: 25 September 2015Accepted: 06 February 2016


BackgroundThe use of the dietary polyphenols as chemosensitizing agents to enhance the efficacy of conventional cytostatic drugs has recently gained the attention of scientists and clinicians as a plausible approach for overcoming the limitations of chemotherapy e.g. drug resistance and cytotoxicity. The aim of this study was to investigate whether a naturally occurring diet-based flavonoid, fisetin, at physiologically attainable concentrations, could act synergistically with clinically achievable doses of paclitaxel to produce growth inhibitory and-or pro-death effects on A549 non-small cell lung cancer cells, and if it does, what mechanisms might be involved.

MethodsThe drug–drug interactions were analyzed based on the combination index method of Chou and Talalay and the data from MTT assays. To provide some insights into the mechanism underlying the synergistic action of fisetin and paclitaxel, selected morphological, biochemical and molecular parameters were examined, including the morphology of cell nuclei and mitotic spindles, the pattern of LC3-II immunostaining, the formation of autophagic vacuoles at the electron and fluorescence microscopic level, the disruption of cell membrane asymmetry-integrity, cell cycle progression and the expression level of LC3-II, Bax, Bcl-2 and caspase-3 mRNA.

ResultsHere, we reported the first experimental evidence for the existence of synergism between fisetin and paclitaxel in the in vitro model of non-small cell lung cancer. This synergism was, at least partially, ascribed to the induction of mitotic catastrophe. The switch from the cytoprotective autophagy to the autophagic cell death was also implicated in the mechanism of the synergistic action of fisetin and paclitaxel in the A549 cells. In addition, we revealed that the synergism between fisetin and paclitaxel was cell line-specific as well as that fisetin synergizes with arsenic trioxide, but not with mitoxantrone and methotrexate in the A549 cells.

ConclusionsOur results provide rationale for further testing of fisetin in the combination with paclitaxel or arsenic trioxide to obtain detailed insights into the mechanism of their synergistic action as well as to evaluate their toxicity towards normal cells in an animal model in vivo. We conclude that this study is potentially interesting for the development of novel chemotherapeutic approach to non-small cell lung cancer.

KeywordsFisetin Paclitaxel Combination therapy Lung cancer Mitotic catastrophe Autophagy AbbreviationsAOacridine orange

AVOsacidic vesicular organelles

Baf A1bafilomycin A1

BSAbovine serum albumin

CIcombination index


Dmthe dose that produces 50 % effect, such as IC50

DMEMDulbecco’s Modified Eagle Medium

fafraction affected the fraction of cells inhibited after the drug exposure, e.g. 0.5 when cell growth is inhibited by 50 %


G6PDglucose 6-phosphate dehydrogenase

HBSSHank’s balanced salt solution

IC50half maximal inhibitory concentration

mthe slope of the median-effect plot, which indicates the shape of the dose–effect curve

MCmitotic catastrophe

MEMMinimum Essential Medium

MPM-2mitotic phosphoprotein monoclonal-2

MTSBmicrotubule stabilizing buffer

MTTthiazolyl blue tetrazolium bromide

NSCLCnon-small cell lung cancer

PBSphosphate-buffered saline

PIpropidium iodide

PSAprostate-specific antigen


qRT-PCRreal-time quantitative reverse transcription PCR

rlinear correlation coefficient, which describes the conformity of the data to the median-effect plot of the mass-action law

RPMI 1640Roswell Park Memorial 1640 medium

RTroom temperature

SCLCsmall-cell lung cancer

SDstandard deviation

TBStris-buffered saline

TEMtransmission electron microscope

TNFtumor necrosis factor

Electronic supplementary materialThe online version of this article doi:10.1186-s12935-016-0288-3 contains supplementary material, which is available to authorized users.

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Autor: Anna Klimaszewska-Wisniewska - Marta Halas-Wisniewska - Tadeusz Tadrowski - Maciej Gagat - Dariusz Grzanka - Alina Grzanka


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