Reactive oxygen species mediate soft corals-derived sinuleptolide-induced antiproliferation and DNA damage in oral cancer cells

Abstract

We previously reported that the soft coral-derived bioactive substance, sinuleptolide, can inhibit the proliferation of oral cancer cells in association with oxidative stress. The functional role of oxidative stress in the cell-killing effect of sinuleptolide on oral cancer cells was not investigated as yet. To address this question, we introduced the reactive oxygen species (ROS) scavenger (N-acetylcysteine [NAC]) in a pretreatment to evaluate the sinuleptolide-induced changes to cell viability, morphology, intracellular ROS, mitochondrial superoxide, apoptosis, and DNA damage of oral cancer cells (Ca9-22). After sinuleptolide treatment, antiproliferation, apoptosis-like morphology, ROS/mitochondrial superoxide generation, annexin V-based apoptosis, and γH2AX-based DNA damage were induced. All these changes were blocked by NAC pretreatment at 4 mM for 1 h. This showed that the cell-killing mechanism of oral cancer cells of sinuleptolide is ROS dependent.

Ethyl acetate extract of Nepenthes adrianii x clipeata induces antiproliferation, apoptosis, and DNA damage against oral cancer cells through oxidative stress

“…MitoSOX Red (Molecular Probes, Invitrogen, Eugene, Oregon) specifically reacted with mitochondrial superoxide (MitoSOX) and generated fluorescence for flow cytometry . Cells were incubated with MitoSOX Red (5 μM) at 37°C for 30 minutes and performed flow cytometry (Accuri C6).…”

A novel sulfonyl chromen‐4‐ones (CHW09) preferentially kills oral cancer cells showing apoptosis, oxidative stress, and DNA damage

“…Moreover, oxidative stress‐generating drugs may induce apoptosis . Accordingly, we found that CHW09 induced ROS and apoptosis of Ca9‐22 oral cancer cells in terms of detection of annexin V and cleavage of PARP and cleavage of caspases 3/8/9, suggesting that both intrinsic and extrinsic apoptosis signaling were involved in CHW09‐induced apoptosis.…”

“…The signal was explored using WesternBright (ECL HRP: #K‐12045‐D50a; Advansta, Menlo Park, CA). An apoptosis inhibitor Z‐VAD‐FMK (http://selleckchem.com; Houston, TX) (25 μM for 2 hr pretreatment) and a free radical scavenger N ‐acetylcysteine (NAC) (Sigma; St. Louis, MO) (2 mM for 1 hr pretreatment) were used to examine the role of apoptosis and oxidative stress in the CHW09 treatment.…”

“…The mitochondrial superoxide was detected using MitoSOX Red (Molecular Probes, Invitrogen, Eugene, OR) for flow cytometry analysis . In brief, drug‐treated cells were incubated with 5 μM MitoSOX at 37°C for 30 min and resuspended in 1 mL PBS to perform flow cytometry (FL3 channel in BD Accuri C6).…”

LY303511 displays antiproliferation potential against oral cancer cells in vitro and in vivo

“…Mitochondrial superoxide interacting dye (MitoSOX Red; Molecular Probes, Invitrogen, Eugene, OR) may generate fluorescence products, which were detectable by flow cytometry . Briefly, cells were treated with MitoSOX Red (5 μM, 37°C) for 30 minutes.…”