title: Anti-proliferative activity of haloperidol in B16 mouse and human SK-MEL-28 melanoma cell lines.
abstract: Sigma receptors are present in cancer cell lines. The aim of the present study is to evaluate the anti-tumor activity of a series of sigma 1, sigma 2 and sigma 1/2 ligands in B16 melanoma cell lines. Proliferation, apoptosis, intracellular ATP content, cell cycle and molecular regulators were analyzed. Cell growth was determined using the sulforhodamine B (SRB) colorimetric cytotoxicity assay. Apoptosis was assessed by flow cytometry and DNA fragmentation, using ELISA cell death assay. ATP content was measured spectrofluorometrically and cell cycle analysis was performed by flow cytometry. The cytoplasmic and nuclear expression of cell cycle regulatory molecules, cyclin D and CDK2 (cyclin dependent kinase 2) were determined by Western blot analysis and quantified by densitometry. The sigma ligands in single digit micromolar concentrations inhibited B16 and multidrug-resistant B16 COL/R cell growth, leading to cell death at higher concentrations. The potency order was: haloperidol, reduced-haloperidol, ifenprodil tartrate, opipramol and carbetapentane citrate. B16 COL/R cells were to some extent, less sensitive to sigma ligands. Further studies have shown that the growth inhibitory effect of sigma ligands could be attributed to G1 arrest of the cell cycle, mediated by a marked decrease in cytoplasmic and nuclear cyclin D and CDK2 protein expression, though haloperidol induced loss of cell viability due to apoptosis. Sigma ligands induced an early decrease in ATP content. These data stimulated us to examine the combined anti-proliferative activity of haloperidol and the tyrosine kinase inhibitor imatinib mesylate (STI 571), on SK-MEL-28 human melanoma cells. Preliminary experiments demonstrated a marked synergistic interaction between the two agents.
authors.affiliation: Endocrinology Laboratory, Rabin Medical Center, Beilinson Campus, Petah Tikva 49100, Israel. jardenan@clalit.org.il
authors.forename: Jardena
authors.initials: J
authors.lastname: Nordenberg
authors.forename: Israela
authors.initials: I
authors.lastname: Perlmutter
authors.forename: Gad
authors.initials: G
authors.lastname: Lavie
authors.forename: Einat
authors.initials: E
authors.lastname: Beery
authors.forename: Orit
authors.initials: O
authors.lastname: Uziel
authors.forename: Chaya
authors.initials: C
authors.lastname: Morgenstern
authors.forename: Eyal
authors.initials: E
authors.lastname: Fenig
authors.forename: Abraham
authors.initials: A
authors.lastname: Weizman
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D001549:Benzamides; D015153:Blotting, Western; D002453:Cell Cycle; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D056741:Cyclin D; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism; D016213:Cyclins / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D053938:DNA Fragmentation; D003720:Densitometry; D018492:Dopamine Antagonists / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006220:Haloperidol / Q000494:pharmacology*; D006801:Humans; D000068877:Imatinib Mesylate; D008024:Ligands; D008055:Lipids; D008545:Melanoma / Q000188:drug therapy* / Q000473:pathology; D008546:Melanoma, Experimental; D051379:Mice; D010879:Piperazines / Q000494:pharmacology; D011485:Protein Binding; D011743:Pyrimidines / Q000494:pharmacology; D012235:Rhodamines / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D013997:Time Factors
chemical_list: D001549:Benzamides; D056741:Cyclin D; D016213:Cyclins; D018492:Dopamine Antagonists; D008024:Ligands; D008055:Lipids; D010879:Piperazines; D011743:Pyrimidines; D012235:Rhodamines; C022027:lissamine rhodamine B; D000068877:Imatinib Mesylate; D000255:Adenosine Triphosphate; D051357:Cyclin-Dependent Kinase 2; D006220:Haloperidol