Molecular response of HL-60 cells to mitotic inhibitors vincristine and taxol visualized with apoptosis-related gene expressions, including the new member BCL2L12.
Taxol and vincristine belong to a group of anticancer drugs that target microtubules, subsequently arresting cells at the mitotic phase of the cell cycle and inducing programmed cell death. The BCL2 (bcl-2) family of genes is of known implication in apoptosis induced by various stimuli, among which BCL2L12, a new member of the family, cloned by our group. For further insights into the mechanisms and molecular targets implicated and modified as a result of apoptosis induced by these two mitosis-arresting drugs, we studied the possible alterations, at the mRNA level, of various apoptosis-related genes (BCL2, BAX, BCL2L12, CASPASE-3, FAS) after leukemia cell (HL-60) treatment with these drugs. The kinetics of cell toxicity were evaluated by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] method, trypan blue staining, and cell proliferation efficiency; apoptosis induction was assayed by endonucleosomal cleavage of DNA (DNA laddering); and the expression levels of the genes were analysed by RT-PCR, using gene-specific primers. The percentage of nonviable cells was upregulated with increasing cell exposure time and drug concentrations to both taxol and vincristine. Distinct modulations of apoptosis-related genes at the mRNA level were also observed, mainly concerning BCL2 and BCL2L12 along apoptosis induction. Our results indicate and support the hypothesis that the apoptosis-related genes BCL2 and BCL2L12 respond similarly to treatment of the human, acute, myelocytic leukemia HL60 cells with the anticancer drugs vincristine and taxol though in a drug-specific and time-dependent manner.
Thomadaki
Hellinida
H
Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, Panepistimiopolis, Athens, Greece.
Floros
Konstantinos V
KV
Scorilas
Andreas
A
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D053148:Caspase 3 / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D009124:Muscle Proteins / Q000235:genetics*; D017239:Paclitaxel / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014750:Vincristine / Q000494:pharmacology*; D051028:bcl-2-Associated X Protein / Q000235:genetics; D019014:fas Receptor / Q000235:genetics
D000972:Antineoplastic Agents, Phytogenic; C430100:BCL2L12 protein, human; C491901:FAS protein, human; D009124:Muscle Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D051028:bcl-2-Associated X Protein; D019014:fas Receptor; D014750:Vincristine; D053148:Caspase 3; D017239:Paclitaxel