Osthole Induces Cell Cycle Arrest and Inhibits Migration and Invasion via PTEN/Akt Pathways in Osteosarcoma

Abstract

Background/Aims: Osteosarcoma is the second highest cause of cancer-related death in children and adolescents. Majority of osteosarcoma patients (90%) show metastasis. Previous reports revealed that osthole showed antitumor activities via induction of apoptosis and inhibition of proliferation. However, the potential effects and detailed molecular mechanisms involved remained unclear. Methods: Cell viability was analyzed by MTT assay in osteosarcoma cell lines MG-63 and SAOS-2. Cell cycle was detected by flow cytometry. The effects of migration and invasion were evaluated by wound healing assay and transwell assays. Moreover, the level of proteins expression was determined by Western blot. Results: The cell viability of MG63 and SAOS-2 were markedly inhibited by osthole in a dose- and time-dependent manner. Cell cycle was arrested and the ability of migration and invasion was obviously reduced when cells were exposed to osthole. Moreover, enzymes involved in PTEN/Akt pathway were regulated such as PTEN and p-Akt proteins. Furthermore, osthole inhibited the tumor growth in vivo. Conclusion: Our study unraveled, for the first time, the ability of osthole to suppress osteosarcoma and elucidated the regulation of PTEN/Akt pathway as a signaling mechanism for the anti-tumor action of osthole. These findings indicate that osthole may represent a novel therapeutic strategy in the treatment of osteosarcoma.

Quercetin Inhibits Cell Migration and Invasion in Human Osteosarcoma Cells

“…Several epidemiological and preclinical studies have highlighted the potential benefits of using flavonoids for cancer prevention. Osthole has been shown to exert anti-cancer effects via the PTEN/Akt signaling pathway [44], and quercetin is one flavonoid whose effects have been widely reported [45-47]. Quercetin exerts its anticancer effects by inducing cell cycle arrest, apoptosis, and differentiation and by suppressing MMP secretion [48].…”

Cyclin-Dependent Kinase Inhibitor p21WAF1/CIP1 Facilitates the Development of Cardiac Hypertrophy

“…Cyclin–CDK complexes are precisely and strictly regulated to prevent abnormal proliferation. There are two families of inhibitors that block the catalytic activity of cyclin-CDK complexes, namely inhibitor of CDK4 (INK4) family which only bind to CDK4/6, and KIP/CIP family including p21 WAF1/CIP1 and p27 KIP1 , which can inhibit all cyclin-CDK complexes [11, 27, 33, 34]. A gene knockout study in mice confirmed that p21 and p27 are important partners in mammalian cardiomyocyte [35].…”

Andrographolide Induces Autophagic Cell Death and Inhibits Invasion and Metastasis of Human Osteosarcoma Cells in An Autophagy-Dependent Manner

“…U-2OS and SaOS-2 cells were cultured in McCoy’s 5A (Sigma, M4892) with 10% fetal bovine serum (CLARK, FB25015), MG-63 cells were cultured in MEM (Gibco 41500-034) with 10% fetal bovine serum, penicillin (100 U/mL) and streptomycin (100 U/mL) in a humidified incubator (Thermo Fisher, Runcorn, Cheshire, UK) with 5% CO 2 at 37°C [22, 23]. …”

“…The reaction was terminated by removing the supernatant, adding dimethyl sulfoxide (150µl/well), and shaking for 10 min at room temperature. The absorbance was measured at 490 nm in a spectrophotometer [22]. …”

“…The confluent U-2OS cells cultured in six-well plates were wounded by yellow tip, cells were received various treatments for 0, 12, 24, 48h, and they were examined under an inverted microscope (Nikon) [22]. …”