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Investigating the Mechanism of Action of Sulforaphane in a Human Leukemia Cell Line
Author(s) -
Fields Peter,
Grom Jennifer,
O'Donnell Robert
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.897.27
Subject(s) - sulforaphane , nocodazole , cell cycle , leukemia , cyclin dependent kinase , cyclin dependent kinase 1 , chemistry , mechanism of action , pharmacology , cancer research , biology , biochemistry , cell , immunology , in vitro , cytoskeleton
Sulforaphane is an isothiocyanate molecule that is found in cruciferous vegetables, such as broccoli. Its anti‐neoplastic properties make it useful as a chemotherapeutic drug. Although it is known to be an effective treatment against leukemia tumor cells, the mechanism by which sulforaphane kills leukemia cells is unknown. Previous labs have shown that sulforaphane effectively arrests leukemia cells in the G2/M cell cycle phase, but what is not clear is whether the phase is G2 or M. In order to clarify the affected phase, roscovitine will be used as a G2 inhibitor and nocodazole will be used as an M phase inhibitor. HL60 leukemia tumor cells will be treated with sulforaphane, roscovitine, and nocodazole, and the results will be compared. Preliminary experiments used sulforaphane, roscovitine, and nocodazole MTT assays to determine the concentration resulting in 75% survival. The concentrations were found to be 4.71 μM, 16.21 μM, and 26.40 nM, respectively. Future experiments will use these doses to compare and identify where the inhibition occurs. Western Blots will be used to look at p21, an inhibitor of several CDK/cyclin complexes, and cdc2 levels (the CDK required to initiate the G2 to M phase transition). Mitotic index and cell cycle analysis will also be used. We hypothesize that if we treat the HL‐60 cells with sulforaphane, there will be an increase in the p21 levels of the cells, supporting arrest in the G2 phase of the cell cycle. We also hypothesize that treatment of leukemia tumor cells with sulforaphane or roscovitine will yield low levels of cdc2 (compared to untreated cells), while nocodazole treated cells will have cdc2 levels that are similar to the untreated cells.