OBJECTIVE To investigate the effects and molecular mechanisms of KZL-188, a novel trifluoromethyl quinazoline derivative, on the proliferation, migration, invasion and apoptosis of osteosarcoma 143B cells.

METHODS The 143B cells were divided into a control group(DMSO) and experimental groups with low, medium and high doses(KZL-188). MTT assay was used to detect the effect of KZL-188 on the proliferation of cells in each group. Flow cytometry was used to detect the effect on apoptosis. Mitochondrial membrane potential test was used to detect the changes of mitochondrial membrane potential. Wound healing assay was used to detect the effect on cell migration. Trans-well assay was used to detect the effect on cell invasion. Western blotting was used to detect the effects of the expression of apoptosis-related proteins of the mitochondrial pathway, metastasis-related proteins, and proteins related to the PI3K/AKT signaling pathway.

RESULTS Compared with the control group, KZL-188 inhibited the proliferation of 143B cells in a concentration-dependent manner; the expression of metastasis-related proteins MMP2, MMP13 and Vimentin decreased in all experimental groups, indicating that KZL-18 could inhibit the migration and invasion of 143B cells. Meanwhile, the mitochondrial membrane potential decreased, and the expression of mitochondrial apoptosis-related proteins Bcl-2 decreased, while the expression of cleaved caspase-3, cleaved caspase-9, Bax and cytochrome C increased. The result indicated that mitochondrial apoptosis occurred in the 143B cells. Further exploration showed that the ratio of p-PI3K/PI3K to p-Akt/Akt related to PI3K/Akt signaling pathway decreased, while the expression level of downstream protein Bim increased. Combined with the PI3K inhibitor LY294002, the inhibitory effect of KZL-188 to PI3K/Akt signaling pathway had been enhanced in 143B cells.

CONCLUSION KZL-188 inhibits the proliferation, migration and invasion of 143B osteosarcoma cells and induces apoptosis of osteosarcoma cells by regulating the expression of mitochondrial apoptosis related proteins, possibly exerting its effects through the PI3K/Akt signaling pathway.