Chrysophanol induces necrosis through the production of ROS and alteration of ATP levels in J5 human liver cancer cells

Abstract

Anthraquinone compounds have been shown to induce apoptosis in different cancer cell types. Effects of chrysophanol, an anthraquinone compound, on cancer cell death have not been well-studied. The goal of this study was to examine if chrysophanol had cytotoxic effects and if such effects involved apoptosis or necrosis in J5 human liver cancer cells. Chrysophanol induced necrosis in J5 cells in a dose- and time-dependent manner. Non-apoptotic cell death was induced by chrysophanol in J5 cells and was characterized by caspase independence, delayed externalization of phosphatidylserine and plasma membrane disruption. Blockage of apoptotic induction by a general caspase inhibitor (z-VAD-fmk) failed to protect cells against chrysophanol-induced cell death. The levels of ROS production and loss of mitochondrial membrane potential (ΔΨm) were also determined to assess the effects of chrysophanol. However, reductions in ATP levels and increases in LDH activity indicated that chrysophanol stimulated necrotic cell death. In summary, human liver cancer cells treated with chrysophanol exhibited a cellular pattern associated with necrosis and not apoptosis.

Gallic acid suppresses the migration and invasion of PC-3 human prostate cancer cells via inhibition of matrix metalloproteinase-2 and -9 signaling pathways

“…5 cells/well of pc-3 cells were cultured in 12-well plates for 24 h, and then various concentrations of GA were added to each well for final concentrations of GA at 25, 50, 100 and 150 µM, and 0.5% DMSO (as a control) for 24 and 48 h. Cells were harvested from each treatment were stained with PI (5 µg/ml), and then analyzed by flow cytometry as previously described (24). These cells were analyzed with a flow cytometer (BD FACSCalibur, San Jose, CA, USA) equipped with an argon ion laser at 488 nm wavelength (24,25).…”

“…These cells were analyzed with a flow cytometer (BD FACSCalibur, San Jose, CA, USA) equipped with an argon ion laser at 488 nm wavelength (24,25).…”

“…The supernatant was collected and total protein was quantitated using a Bio-Rad protein assay kit (Hercules, CA, USA) with bovine serum albumin (BSA) as the standard. Protein abundance of MMP-2, MMP-9, SOS1, GRB2, PKC, NF-κB p65, JNK1/2, ERK1/2, p38, PI3K, AKT, p-AKT (Thr308) and p-AKT (Ser473) (Santa Cruz Biotechnology, Inc.) were determined by SDS-PAGE and Western blotting as previously described (24)(25)(26). NIH Image J software was used to determine the band intensity from Western blot analysis.…”

“…6 cells/well) were placed in 6-well plates and incubated without and with GA (25 or 50 µM) for 24 h. Cells were harvested and total RNA was extracted from each treatment by using the Qiagen RNeasy Mini Kit (Qiagen, Inc., valencia, CA, USA) as described previously (24,25). RNA samples were reverse-transcribed at 42˚C with High Capacity cDNA Reverse Transcription Kit for 30 min according to the protocol of the supplier (Applied Biosystems, Carlsbad, CA, USA).…”

A novel synthetic 2-(3-methoxyphenyl)-6,7-methylenedioxoquinolin-4-one arrests the G2/M phase arrest via Cdc25c and induces apoptosis through caspase- and mitochondria-dependent pathways in TSGH8301 human bladder cancer cells

“…The fixed cells were then incubated with anti-human AIF, Endo G and cytochrome c primary antibody (1:100 dilution) overnight and then exposed to the secondary antibody (FITC-conjugated goat anti-mouse IgG at 1:100 dilution), followed by DNA staining with PI. The cells in each slide were then photomicrographed by using a Leica TCS SP2 Confocal Spectral Microscope as described previously (19)(20)(21) …”