Evodiamine Induces Apoptosis, G2/M Cell Cycle Arrest, and Inhibition of Cell Migration and Invasion in Human Osteosarcoma Cells via Raf/MEK/ERK Signalling Pathway

Abstract

BackgroundOsteosarcoma is a prevalent type of bone tumor mainly reported in children and adolescents. The treatments for osteosarcoma are limited and are associated with serious adverse effects. In this study we evaluated the anticancer activity of Evodiamine, a plant-derived natural product, against a panel of osteosarcoma cells and explored the underlying mechanisms.Material/MethodsThe viability of osteosarcoma cell lines was investigated by MTT assay. Apoptosis was detected by DAPI and annexin V/PI staining and cell cycle analysis was performed by flow cytometry. The expression of the proteins was examined by Western blotting.ResultsThe results of the present study indicated that Evodiamine inhibited the proliferation of U2OS osteosarcoma cells with an IC50 of 6 μM. Further investigations indicated the antiproliferative effects of Evodiamine are due to induction of apoptosis and G2/M cell cycle arrest. The results of Western blotting revealed that the expression of several apoptosis (Cytochrome c, Bax, Bid, Caspase 3, 9, 8, and PARP) and cell cycle-related proteins (cyclin B1, Cdc25c, and Cdc2) was significantly altered. Evodiamine also suppressed the migration and invasion of U2OS osteosarcoma cells. Moreover, Evodiamine downregulated the expression of important regulatory proteins such as p-MEK and p-ERK, leading to the inhibition of Raf/MEK/ERK signalling pathways.ConclusionsWe found that Evodiamine exerts anticancer effects on osteosarcoma cells and has potential in the treatment of osteosarcoma.

Identification of key biomarkers and functional pathways in osteosarcomas with lung metastasis: Evidence from bioinformatics analysis

“…encoded cell cycle regulatory protein that participated in regulating G2/M progression and mediating DNA damage repair. It has been reported that Evodiamine, Flavokawain B and Ludartin could inhibit proliferation, migration, and apoptosis of osteosarcoma cells by down-regulating the expression level of CDC25C [62][63][64] . At present, the relationship between the rest of hub genes (DTL, CENPN, PARPBP, SMC2) and osteosarcoma remains unclear and the speci c role in lung metastasis deserves further exploration.…”

Antiproliferative Effects of Alkaloid Evodiamine and Its Derivatives

“…Evodiamine was shown to inhibit the proliferation of human osteosarcoma 143B02 cells through inactivation of PTEN/PI3K/Akt pathway [ 154 ]. By inactivating Raf/MEK/ERK signaling pathway, evodiamine also induced growth inhibition and inactivated the activities of migration and invasion of U2OS cells [ 155 , 156 ]. Moreover, the inactivation of Wnt/β-catenin signaling pathway contributed to the cytotoxic activity of evodiamine in human osteosarcoma MG-63 cells [ 157 ].…”

RAF1 as Downstream Molecule Mediates the FSH Signaling Pathway to Stimulate E2 Synthesis and Secretion in Mouse Ovarian Granulosa Cells

“…Many studies suggested that RAF can affect the proliferation of cell through in uencing the distribution of cell cycle [29][30][31], and we speculated that the signi cant and rapid change of estradiol level was caused by GCs proliferation. Our ow cytometry analysis results showed that RAF709 did not induce the suppression of cell cycle stage, but affected on the activity of estradiol synthase, which con rmed that RAF1 could regulate the expression of CYP19A1 [32] through the ERK signaling pathways [33].…”