Suppression of autophagy enhances preferential toxicity of epothilone A and epothilone B in ovarian cancer cells.

BACKGROUND

Epothilones are microtubule-targeting agents that induce death in a variety of cancer cell types. Here, we focus on the cellular and molecular mechanisms underlying epothilone A (Epo A) and epothilone B (Epo B)-induced autophagy and apoptosis in ovarian cancer cells, compared to the actions of the widely used clinical chemotherapy drug paclitaxel (PTX).

MATERIALS AND

METHODS

Autophagy was examined in two cell lines, SKOV-3 (human ovarian adenocarcinoma) and OV-90 (human ovarian papillary serous adenocarcinoma), which differ in the levels of p-glycoprotein and drug resistance, based on the LC3 ELISA assay, fluorescence detection of autophagosome formation, morphological changes evaluated via acridine orange staining, and visualization of LC3 protein using confocal microscopy. Cell viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. Apoptosis was measured via the caspase-3/7 assay and immunofluorescence labeling of caspase-3. Differences in microtubule organization in epothilone-treated cells were investigated using specific antibodies against β-tubulin. All probes were analyzed both in the presence and absence of the autophagy inhibitor, bafilomycin A1 (Baf), and apoptosis inhibitor, Z-FA-FMK.

RESULTS

Epothilone and PTX treatment induced a dose-dependent decrease in cell viability, along with increased apoptosis and disruption of microtubule dynamics. Furthermore, under conditions of inhibition of autophagy with Baf, apoptosis triggered by these compounds was significantly increased.

CONCLUSION

Our collective results suggest that treatment with epothilones in combination with autophagy inhibitors present a potentially more effective chemotherapeutic approach for ovarian cancer.