Propyl-2-(8-(3,4-Difluorobenzyl)-2′,5′-Dioxo-8-Azaspiro[Bicyclo[3.2.1] Octane-3,4′-Imidazolidine]-1′-yl) Acetate Induces Apoptosis in Human Leukemia Cells through Mitochondrial Pathway following Cell Cycle Arrest

Abstract

BackgroundDue to the functional defects in apoptosis signaling molecules or deficient activation of apoptosis pathways, leukemia has become an aggressive disease with poor prognosis. Although the majority of leukemia patients initially respond to chemotherapy, relapse is still the leading cause of death. Hence targeting apoptosis pathway would be a promising strategy for the improved treatment of leukemia. Hydantoin derivatives possess a wide range of important biological and pharmacological properties including anticancer properties. Here we investigated the antileukemic activity and mechanism of action of one of the potent azaspiro hydantoin derivative, (ASHD).Materials and MethodsTo investigate the antileukemic efficacy of ASHD, we have used MTT assay, cell cycle analysis by FACS, tritiated thymidine incorporation assay, Annexin V staining, JC1 staining and western blot analysis.ResultsResults showed that ASHD was approximately 3-fold more potent than the parent compounds in inducing cytotoxicity. Tritiated thymidine assay in conjunction with cell cycle analysis suggests that ASHD inhibited the growth of leukemic cells. The limited effect of ASHD on cell viability of normal cells indicated that it may be specifically directed to cancer cells. Translocation of phosphatidyl serine, activation of caspase 3, caspase 9, PARP, alteration in the ratio of BCL2/BAD protein expression as well as the loss of mitochondrial membrane potential suggests activation of the intrinsic pathway of apoptosis.ConclusionThese results could facilitate the future development of novel hydantoin derivatives as chemotherapeutic agents for leukemia.

Purification and Characterization of Glutaminase Free Asparaginase from Enterobacter cloacae: In-Vitro Evaluation of Cytotoxic Potential against Human Myeloid Leukemia HL-60 Cells

“…cloacae asparaginase, HL-60 cells showed all aforementioned morphological changes, which were increased in dose-dependent manner. Similar observations were also recorded with asparaginase treated mouse lymphoma [ 49 ] and human T lymphocytes cells [ 50 ] and with other molecules recently tested as potent antileukemic agent [ 51 , 52 ].…”

“…Mitochondrial membrane potential (MMP or Δψm) loss is an essential event of apoptosis [ 54 ] and the measurement of MMP loss is one of the methods used for investigation of apoptosis [ 51 ]. Hence, further we attempted to understand the effect of purified enzyme on mitochondrial membrane permeability in HL-60 cells.…”

Autocyclized and oxidized forms ofSCR7 induce cancer cell death by inhibiting nonhomologousDNAend joining in a LigaseIVdependent manner

“…The NHEJ assay was performed as described before . Briefly, cell‐free extracts prepared from rat testes were incubated with or without SCR7‐cyclized or SCR7‐pyrazine (100, 200, 500 μ m and 1 m m ) for 30 min at 25 °C in NHEJ buffer [25 m m Tris‐HCl, (pH 7.5), 75 m m NaCl, 10 m m MgCl 2 , 42.5 m m KCl, 0.025% Triton X‐100, 100 μg·mL −1 BSA, 10% PEG, and 5% glycerol].…”

Identification of a novel BCL2‐specific inhibitor that binds predominantly to the BH1 domain

“…To check the mode of cell death induced by the Disarib, annexin‐FITC/PI staining was carried out as described previously . Briefly, Molt4 cells (0.25 × 10 5 cells) were treated with Disarib (0, 1, 2, and 5 μ m ) for 48 h. DMSO‐treated cells were used as control.…”