Enhanced G2/M Arrest, Caspase Related Apoptosis and Reduced E-Cadherin Dependent Intercellular Adhesion by Trabectedin in Prostate Cancer Stem Cells.
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.
Department of Histology and Embryology, Ege University Faculty of Medicine, Bornova, Izmir, Turkey.
Eda
E
Acikgoz
Department of Stem Cell, Ege University Health Science Institute, Bornova, Izmir, Turkey.
Ummu
U
Guven
Department of Stem Cell, Ege University Health Science Institute, Bornova, Izmir, Turkey.
Fahriye
F
Duzagac
Department of Medical Oncology, Ege University Faculty of Medicine, Bornova, Izmir, Turkey.
Ruchan
R
Uslu
Department of Histology and Embryology, Yuzuncu Yil University Faculty of Medicine, Van, Turkey.
Mikail
M
Kara
Department of Medical Pharmacology, Necmettin Erbakan University, Meram Faculty of Medicine, Konya, Turkey.
Burak Cem
BC
Soner
Department of Histology and Embryology, Ege University Faculty of Medicine, Bornova, Izmir, Turkey.
Gulperi
G
Oktem
D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D015820:Cadherins / Q000378:metabolism*; D020169:Caspases / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D003114:Colony-Forming Units Assay; D004149:Dioxoles / Q000494:pharmacology*; D005455:Fluorescent Antibody Technique; D016195:G2 Phase / Q000187:drug effects*; D006801:Humans; D008297:Male; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D011471:Prostatic Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D044005:Tetrahydroisoquinolines / Q000494:pharmacology*; D000077606:Trabectedin; D014407:Tumor Cells, Cultured
D018906:Antineoplastic Agents, Alkylating; D015820:Cadherins; D004149:Dioxoles; D044005:Tetrahydroisoquinolines; D020169:Caspases; D000077606:Trabectedin