LY404039 Inhibits Proliferation and Metastasis of Osteosarcoma Cells via PI3K Signaling Pathway.

ABSTRACT

Osteosarcoma is one of the most aggressive primary malignant bone tumors, and the effect of first-line therapeutic schedule had no improvement in past few years. Therefore, it is extremely urgent to develop new drugs against osteosarcoma. In this study, human osteosarcoma U2 OS cells as a model in vitro were treated with LY404039 and dimethyl sulfoxide. CCK-8 proliferation test, transwell assay, and flow cytometry were used to assess cell proliferation, invasion/migration, and apoptosis, respectively. Moreover, western blotting was performed to determine the relative expression of protein related to apoptosis and PI3K signaling pathway. The proliferation of U2 OS cells was significantly inhibited 72 h after LY404039-treatment by CCK-8 proliferation test. Matrigel invasion analysis showed the numbers of invasive cells were significantly decreased in LY404039-treated group compared with the NC group. Furthermore, LY404039 led to the remarkable reduction of the number of U2 OS cells passing through the microwells of the transwell chamber. The expression levels of anti-apoptotic protein BCL-2 decreased, and the expression of pro-apoptotic proteins Active Caspase3, Bax increased concurrently after LY404039 stimulation using western blot. The phosphorylation of AKT and mTOR were significantly inhibited in U2 OS cells treated with LY404039. Also, the activity of the downstream proteins such as p70S6K and Cyclin D1 were also significantly decreased. LY404039 could inhibit proliferation and metastasis of osteosarcoma cells, which might be attributed to the enhancement of apoptosis via PI3K pathway.