Journal Article

Enhanced G2/M Arrest, Caspase Related Apoptosis and Reduced E-Cadherin Dependent Intercellular Adhesion by Trabectedin in Prostate Cancer Stem Cells

Kara, Mikail, Uslu, Ruchan, Oktem, Gulperi, Soner, Burak Cem, Guven, Ummu, Duzagac, Fahriye, Acikgoz, Eda

2015

Overview

Trabectedin (Yondelis, ET-743) is a marine-derived tetrahydroisoquinoline alkaloid. It is originally derived from the Caribbean marine tunicate Ecteinascidia turbinata and currently produced synthetically. Trabectedin is active against a variety of tumor cell lines growing in culture. The present study focused on the effect of trabectedin in cell proliferation, cell cycle progression, apoptosis and spheroid formation in prostate cancer stem cells (CSCs). Cluster of differentiation (CD) 133+high/CD44+high prostate CSCs were isolated from the DU145 and PC-3 human prostate cancer cell line through flow cytometry. We studied the growth-inhibitory effects of trabectedin and its molecular mechanisms on human prostate CSCs and non-CSCs. DU-145 and PC-3 CSCs were treated with 0.1, 1, 10 and 100 nM trabectedin for 24, 48 and 72 h and the growth inhibition rates were examined using the sphere-forming assay. Annexin-V assay and immunofluorescence analyses were performed for the detection of the cell death. Concentration-dependent effects of trabectedin on the cell cycle were also evaluated. The cells were exposed to the different doses of trabectedin for 24, 48 and 72 h to evaluate the effect of trabectedin on the number and diameter of spheroids. According to the results, trabectedin induced cytotoxicity and apoptosis at the IC50 dose, resulting in a significant increase expression of caspase-3, caspase-8, caspase-9, p53 and decrease expression of bcl-2 in dose-dependent manner. Cell cycle analyses revealed that trabectedin induces dose-dependent G2/M-phase cell cycle arrest, particularly at high-dose treatments. Three-dimensional culture studies showed that trabectedin reduced the number and diameter of spheroids of DU145 and PC3 CSCs. Furthermore, we have found that trabectedin disrupted cell-cell interactions via E-cadherin in prostasphere of DU-145 and PC-3 CSCs. Our results showed that trabectedin inhibits cellular proliferation and accelerates apoptotic events in prostate CSCs; and may be a potential effective therapeutic agent against prostate cancer.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Analysis / Antineoplastic Agents, Alkylating - pharmacology / Apoptosis / Apoptosis - drug effects / Bcl-2 protein / Breast cancer / Cadherins - metabolism / Cancer / Cancer therapies / Care and treatment / Caspase / Caspase-3 / Caspase-8 / Caspase-9 / Caspases - metabolism / CD44 antigen / Cell adhesion / Cell Adhesion - drug effects / Cell culture / Cell cycle / Cell death / Cell division / Cell Division - drug effects / Cell interactions / Cell proliferation / Cell Proliferation - drug effects / Chemical compounds / Chemotherapy / Colony-Forming Units Assay / Cytochrome / Cytometry / Cytotoxicity / Deoxyribonucleic acid / Development and progression / Dioxoles - pharmacology / DNA / DNA repair / Drug resistance / E-cadherin / Embryology / Flow cytometry / Fluorescent Antibody Technique / G2 Phase - drug effects / Histology / Humans / Immunofluorescence / Life assessment / Male / Medicine / Metastasis / Molecular modelling / Neoplastic Stem Cells - drug effects / Neoplastic Stem Cells - metabolism / Neoplastic Stem Cells - pathology / Ovarian cancer / p53 Protein / Pharmacology / Physiological aspects / Prostate cancer / Prostatic Neoplasms - drug therapy / Prostatic Neoplasms - metabolism / Prostatic Neoplasms - pathology / Signal transduction / Spheroids / Stem cells / Tetrahydroisoquinoline / Tetrahydroisoquinolines - pharmacology / Toxicity / Tumor cell lines / Tumor Cells, Cultured / Tumor proteins / Tumors / University faculty