Solanum lyratumExtracts Induce Extrinsic and Intrinsic Pathways of Apoptosis in WEHI-3 Murine Leukemia Cells and Inhibit Allograft Tumor

Abstract

<jats:p>We investigated the molecular mechanisms of cell cycle arrest and apoptotic death induced by<jats:italic>Solanum lyratum</jats:italic>extracts (SLE) or diosgenin in WEHI-3 murine leukemia cells<jats:italic>in vitro</jats:italic>and antitumor activity<jats:italic>in vivo</jats:italic>. Diosgenin is one of the components of SLE. Our study showed that SLE and diosgenin decreased the viable WEHI-3 cells and induced<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mrow><mml:msub><mml:mtext>G</mml:mtext><mml:mtext>0</mml:mtext></mml:msub><mml:mo>/</mml:mo><mml:msub><mml:mtext>G</mml:mtext><mml:mtext>1</mml:mtext></mml:msub></mml:mrow></mml:math>phase arrest and apoptosis in concentration- or time-dependent manners. Both reagents increased the levels of ROS production and decreased the mitochondrial membrane potential (<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mrow><mml:mi>Δ</mml:mi><mml:msub><mml:mi>Ψ</mml:mi><mml:mi>m</mml:mi></mml:msub></mml:mrow></mml:math>). SLE- and diosgenin-triggered apoptosis is mediated through modulating the extrinsic and intrinsic signaling pathways. Intriguingly, the p53 inhibitor (pifithrin-α), anti-Fas ligand (FasL) mAb, and specific inhibitors of caspase-8 (z-IETD-fmk), caspase-9 (z-LEHD-fmk), and caspase-3 (z-DEVD-fmk) blocked SLE- and diosgenin-reduced cell viability of WEHI-3 cells. The<jats:italic>in vivo</jats:italic>study demonstrated that SLE has marked antitumor efficacy against tumors in the WEHI-3 cell allograft model. In conclusion, SLE- and diosgenin-induced<mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML"><mml:mrow><mml:msub><mml:mtext>G</mml:mtext><mml:mtext>0</mml:mtext></mml:msub><mml:mo>/</mml:mo><mml:msub><mml:mtext>G</mml:mtext><mml:mtext>1</mml:mtext></mml:msub></mml:mrow></mml:math>phase arrest and triggered extrinsic and intrinsic apoptotic pathways via p53 activation in WEHI-3 cells. SLE also exhibited antitumor activity<jats:italic>in vivo</jats:italic>. Our findings showed that SLE may be potentially efficacious in the treatment of leukemia in the future.</jats:p>

In vivo evaluation of the synthesized novel 2-benzyloxybenzaldehyde analog CCY-1a-E2 for the treatment of leukemia in the BALB/c mouse WEHI-3 allograft model

“…The murine allograft model is frequently used for experimental antileukemic therapy as it is quick and easy to induce leukemia (13,19). The murine WEHI-3 myelomonocytic leukemia cell line originally derived from the BALB/c mouse was first established in 1969 and it has been used to induce leukemia in BALB/c mice for evaluating the antileukemic effects of drugs (13,20,21).…”

Bufalin inhibits migration and invasion in human hepatocellular carcinoma SK-Hep1 cells through the inhibitions of NF-kB and matrix metalloproteinase-2/-9-signaling pathways

“…SK‐Hep1 cells at a density of 5 × 10 4 cells/well were placed in four‐well chamber slides and treated with bufalin (0 and 10 nM) for 24 h. Cells were then fixed in 3% formaldehyde in PBS for 15 min, permeabilized with 0.1% Triton X‐100 in PBS for 1 h with the blocking of nonspecific binding sites using 2% BSA as previously described (Yang et al, ). Cells were stained with antiNF‐κB (1:200 dilution as a primary antibody) overnight and then were stained with FITC‐conjugated goat antimouse IgG at 1:100 dilution (as the secondary antibody) (green fluorescence), followed by nuclei counterstaining with PI (red fluorescence).…”

“…Cells were stained with antiNF‐κB (1:200 dilution as a primary antibody) overnight and then were stained with FITC‐conjugated goat antimouse IgG at 1:100 dilution (as the secondary antibody) (green fluorescence), followed by nuclei counterstaining with PI (red fluorescence). Cells were then photomicrographed using a Leica TCS SP2 confocal spectral microscope (Yang et al, ).…”

Ethanol extract ofHedyotis diffusawilld upregulates G0/G1 phase arrest and induces apoptosis in human leukemia cells by modulating caspase cascade signaling and altering associated genes expression was assayed by cDNA microarray

“…Assays for Caspase-3, Caspase-8, and Caspase-9 Activities HL-60 cells in 12-well plates were incubated in the presence or absence of 100 lg/mL of EEHDW for 24 h. Cells were then harvested, and cell lysates were assessed in accordance with the manufacturer's instruction provided in the caspase-3, caspase-8, and caspase-9 Colorimetric Assay Kits (R&D System). Cell lysates containing 50 lg proteins were incubated for 1 h at 37 C with individual specific caspase-3, caspase-8, or caspase-9 substrate in the reaction buffer (provided in the kits) and measured by OD 405 of the released DNA as previously described (Chiu et al, 2011;Huang et al, 2012;Yang et al, 2012).…”