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

Abstract

Oral cancer is a serious and fatal disease. Cisplatin is the first line of chemotherapeutic agent for oral cancer therapy. However, the development of drug resistance and severe side effects cause tremendous problems clinically. In this study, we investigated the pharmacologic mechanisms of YC-1 on cisplatin-resistant human oral cancer cell line, CAR. Our results indicated that YC-1 induced a concentration-dependent and time-dependent decrease in viability of CAR cells analyzed by MTT assay. Real-time image analysis of CAR cells by IncuCyte™ Kinetic Live Cell Imaging System demonstrated that YC-1 inhibited cell proliferation and reduced cell confluence in a time-dependent manner. Results from flow cytometric analysis revealed that YC-1 promoted G0/G1 phase arrest and provoked apoptosis in CAR cells. The effects of cell cycle arrest by YC-1 were further supported by up-regulation of p21 and down-regulation of cyclin A, D, E and CDK2 protein levels. TUNEL staining showed that YC-1 caused DNA fragmentation, a late stage feature of apoptosis. In addition, YC-1 increased the activities of caspase-9 and caspase-3, disrupted the mitochondrial membrane potential (AYm) and stimulated ROS production in CAR cells. The protein levels of cytochrome c, Bax and Bak were elevated while Bcl-2 protein expression was attenuated in YC-1-treated CAR cells. In summary, YC-1 suppressed the viability of cisplatin-resistant CAR cells through inhibiting cell proliferation, arresting cell cycle at G0/G1 phase and triggering mitochondria-mediated apoptosis. Our results provide evidences to support the potentially therapeutic application of YC-1 on fighting against drug resistant oral cancer in the future.

Ouabain induces apoptotic cell death in human prostate DU 145 cancer cells through DNA damage and TRAIL pathways

“…And then cells were collected, counted and stained with PI (5 μg/mL) for total cell viability determination by using flow cytometric assay (BD Biosciences, FACSCalibur, San Jose, California). Cells were treated with 10 μM of ouabain for 0, 6, 24, and 48 hours for cell cycle distribution assay by flow cytometry as previously described …”

“…Cells were treated with 10 μM of ouabain for 0, 6, 24, and 48 hours for cell cycle distribution assay by flow cytometry as previously described. 37,38 2.4 | Annexin V/PI staining for examination of apoptotic cell death DU 145 cells (1 × 10 5 cells/well) in 12-well culture plates were treated with 10 μM of ouabain for 48 hours. Then, cells were harvested, resuspended in Annexin V binding buffer, followed incubated with Annexin V-FITC/PI in the dark for 15 minutes.…”

Fisetin Induces Apoptosis of HSC3 Human Oral Cancer Cells Through Endoplasmic Reticulum Stress and Dysfunction of Mitochondria-mediated Signaling Pathways

“…For ΔΨ m measurement, cells were isolated and re-suspended with 500 μl of 3,3'-dihexyloxacarbocyanine iodide (DiOC 6 ) (Sigma Chemical Co.) (4 μmol/l) and maintained in the dark for 30 minutes. After incubation, all samples were analyzed by flow cytometry as described previously (33)(34)(35). in vivo 31: 1103-1114 (2017) Measurements of caspase activities.…”

Combinational treatment of all‐trans retinoic acid (ATRA) and bisdemethoxycurcumin (BDMC)‐induced apoptosis in liver cancer Hep3B cells

“…Subsequently, cells were determined the distribution of cell cycle by flow cytometry (Ni et al, 2012). Or the apoptosis rate was analyzed by FITC-Annexin V/PI kit (BD PharMingen; San Diego, CA, USA) according to the manufacturer's instructions (Lee et al, 2017;Ni et al, 2012).…”