Journal Article

The roles of bioactive sphingolipids in resveratrol-induced apoptosis in HL60 acute myeloid leukemia cells

Cakir, Zeynep, Saydam, Guray, Baran, Yusuf, Sahin, Fahri

2011

Overview

Purpose Acute promyelocytic leukemia results from a translocation between 15 and 17 chromosomes that produce PML/RARα fusion protein. PML/RARα inhibits differentiation of myeloid precursor cells at stem cell level. Resveratrol is a phytoalexin that exerts cytotoxic effects on cancer cells. Ceramides have crucial roles in cell growth, proliferation, differentiation, drug resistance, and apoptosis. In this study, we examined the possible cytotoxic effects of resveratrol on acute myeloid leukemia cells and determined the roles of ceramide-metabolizing genes in resveratrol-induced apoptosis, in addition to investigating the possibility of increasing the sensitivity of HL60 cells to resveratrol by manipulating sphingolipids. Methods Cytotoxic effects of resveratrol, C8:ceramide, PDMP, and SK-1 inhibitor were determined by XTT cell proliferation assay. Changes in caspase-3 enzyme activity and mitochondrial membrane potential (MMP) were measured using caspase-3 colorimetric assay and JC-1 MMP detection kit. Expression levels of ceramide-metabolizing genes were examined by RT-PCR. Results The results revealed that manipulations of ceramide metabolism toward generation or accumulation of apoptotic ceramides increased apoptotic effects of resveratrol in HL60 cells, synergistically. More importantly, gene expression analyses revealed that resveratrol-induced apoptosis via increasing expression levels of ceramide-generating genes and decreasing expression levels of antiapoptotic sphingosine kinase-1 and glucosylceramide synthase genes. Conclusion These results showed for the first time that increasing intracellular levels of ceramides by biochemical approaches has also increased sensitivity of HL60 cells to resveratrol. We also showed that resveratrol induces apoptosis through manipulating ceramide-metabolizing genes that resulted in the accumulation of ceramides in HL60 cells.

• Publisher: Springer-Verlag,Springer,Springer Nature B.V
• Subject: Acute myeloid leukemia / Acute promyeloid leukemia / Amino Alcohols - pharmacology / Antineoplastic agents / Antineoplastic Agents, Phytogenic - pharmacology / Antioxidants - pharmacology / Apoptosis / Apoptosis - drug effects / Apoptosis - genetics / Biological and medical sciences / Cancer Research / Caspase 3 - metabolism / Caspase-3 / Cell differentiation / Cell proliferation / Cell Proliferation - drug effects / Ceramide / Ceramide glucosyltransferase / Ceramides - pharmacology / Colorimetry / Cytotoxicity / Drug resistance / Drug Synergism / Enzymatic activity / Enzymes / Fusion protein / Gene expression / Gene Expression Regulation, Neoplastic - drug effects / Hematologic and hematopoietic diseases / Hematology / HL-60 Cells - drug effects / HL-60 Cells - metabolism / Humans / Internal Medicine / Intracellular levels / Leukemia / Leukemia, Myeloid, Acute - drug therapy / Leukemia, Myeloid, Acute - metabolism / Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis / Lipids / Medical sciences / Medicine / Medicine & Public Health / Membrane potential / Membrane Potential, Mitochondrial - drug effects / Membrane Proteins - metabolism / Morpholines - pharmacology / Neoplasm Proteins - drug effects / Neoplasm Proteins - genetics / Neoplasm Proteins - metabolism / Oncology / Original Paper / Pharmacology. Drug treatments / Polyphenols / Promyeloid leukemia / Resveratrol / Reverse Transcriptase Polymerase Chain Reaction / Sphingolipids / Sphingolipids - metabolism / Sphingosine kinase / Stilbenes - pharmacology / Tumor Suppressor Proteins - metabolism