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  • Title: Molecular response of HL-60 cells to mitotic inhibitors vincristine and taxol visualized with apoptosis-related gene expressions, including the new member BCL2L12.
    Author: Thomadaki H, Floros KV, Scorilas A.
    Journal: Ann N Y Acad Sci; 2009 Aug; 1171():276-83. PubMed ID: 19723066.
    Abstract:
    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.
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