Novel quinazoline HMJ‐30 induces U‐2 OS human osteogenic sarcoma cell apoptosis through induction of oxidative stress and up‐regulation of ATM/p53 signaling pathway

Abstract

Human osteogenic sarcoma is the most common primary bone tumor. Despite of the success of frontline therapy, about 40% of patients have disease progression and further therapy is palliative and toxic. In this study, we developed a novel quinazoline HMJ-30 to investigate the cell growth inhibition and apoptotic responses in U-2 OS human osteogenic sarcoma cells. Our results demonstrated that HMJ-30 significantly reduced cell viabilities of U-2 OS, HOS, and 143B cells in a dose-dependent manner, but it exhibited low cytotoxicity in normal hFOB cells. HMJ-30 induced DNA damage and apoptosis in U-2 OS cells as revealed by morphologic changes, comet assay and DAPI staining. Immuno-staining, colorimetric assays, and Western blotting analyses indicated that activities of caspase-8, caspase-9, and caspase-3 and the levels of Bcl-2 family-related proteins (Bcl-2, Mcl-1, Bax, BAD, and t-Bid) were altered in HMJ-30-treated U-2 OS cells. Pretreatment of cells with caspase-8, -9, and -3 specific inhibitors significantly reduced the cell growth inhibition. HMJ-30-induced apoptosis was mediated through both death-receptor and mitochondria-dependent apoptotic pathways in U-2 OS cells. HMJ-30 induced early phosphorylation of p53(Ser18) was through the activation of ataxia telangiectasia mutated (ATM) in U-2 OS cells. The cell growth inhibition by HMJ-30 was substantially attenuated either by the pre-incubation of U-2 OS cells with N-acetylcysteine (NAC, an antioxidant) and caffeine (an ATM kinase inhibitor) or by p53 knockdown via RNAi. In conclusion, ROS dependent-ATM/p53 signaling pathway is involved in HMJ-30-induced apoptosis in U-2 OS cells.

Kaempferol induces ATM/p53-mediated death receptor and mitochondrial apoptosis in human umbilical vein endothelial cells

“…It was reported that ROS can modulate death receptor pathway in cancer cells (8,10). Our hypothesis showed that extrinsic death receptor pathway is a central component in kaempferol-triggered apoptosis of HUVECs.…”

“…The agarose-cell mixture (50 µl) was immediately pipetted onto comet slides. The slides were then immersed in pre-chilled lysis solution for 30 min at 4˚C as previously described (10). After lysis, horizontal electrophoresis was performed for 30 min at 300 mA.…”

“…p53 phosphorylation on the residue of Ser15 has been linked to apoptosis and shown to be a transcription factor to modulate apoptotic target genes such as Fas and DR5 (9). p53 gene expression has been shown to upregulate both the extrinsic and the intrinsic apoptotic signaling pathways (6,10).…”

YC-1 induces G0/G1phase arrest and mitochondria-dependent apoptosis in cisplatin-resistant human oral cancer CAR cells

“…CAR cells (2 × 10 5 cells/per well) were plated into the 12-well plates and then treated with 100 μM of YC-1 for 0, 12, 24, 36 and 48 h. The cells were then fixed, followed by staining with propidium iodide (PI) solution as previously described [91, 92]. The cell cycle profiling and the data analysis were determined utilizing a Muse Cell Analyzer (Merck Millipore, Hayward, CA, USA) [93–98].…”

Kaempferol induces autophagy through AMPK and AKT signaling molecules and causes G2/M arrest via downregulation of CDK1/cyclin B in SK-HEP-1 human hepatic cancer cells

“…The concentration of pNA released from the substrate was calculated using a calibration curve prepared from defined pNA solutions. The experiments were performed in triplicate (27).…”