To further exclude the possibility that the HGF that had below been secreted before serum starvation could have bound the c Met receptor and triggered con stitutive c Met phosphorylation, PC 3 cells were quickly rinsed with a wash buffer to strip any potential pre exist ing HGF molecules on the cell surface. The results showed that even after the rinse, the expression of p c Met and p Akt still remained unchanged. PC 3 was responsive to the small molecule Met kinase inhibitor BMS 777607 To test whether a small molecule Met kinase inhibitor could impair critical Met associated cell functions, PC 3 cells were exposed to BMS 777607. Both cell proliferation found to be significantly inhibited by BMS 777607 at 1 uM.

Anoikis is a mode of anchorage independent cell death that negatively affects cancer cell dissemination and anoikis resistance is considered as a critical Inhibitors,Modulators,Libraries player Inhibitors,Modulators,Libraries in prostate cancer metastasis. To test whether Met inhibition will lead to anoikis, suspended PC 3 cells were incubated with BMS 777607 or wortmannin for 3 days. While wortmannin significantly increased anchorage independent cell death, BMS 777607 did not significantly affect anoikis even at the highest dose tested. BMS 777607 blocked constitutive c Met signaling in PC 3 cells To investigate signaling Inhibitors,Modulators,Libraries alterations after c Met kinase inhibition, cells were exposed to BMS 777607 for vari ous doses and times. BMS 777607 completely eliminated c Met autophosphorylation at doses as low as 0. 1 uM. While p Akt was modestly inhibited by BMS 777607 at the highest dose, expression levels of autophosphorylated Src and Src dependent phosphorylated FAK were decreased with doses greater than 0.

5 uM. In contrast, autophosphorylated FAK was not affected by BMS 777607. When cells were treated with BMS 777607 for prolonged periods, phosphoryl ation of c Met, c Src and FAK remained inhibited. Furthermore, phosphorylation of Akt and mammalian tar Inhibitors,Modulators,Libraries get of rapamycin as well as downstream mole cules S6K and S6 started to be ablated at 3 24 h after drug treatment. Inhibitors,Modulators,Libraries ERK phos phorylation however, showed little change by either high dose or long term treatment. and clonogenicity were found to be impaired by BMS 777607 with doses greater than 1 uM. However, apoptosis was not observed even with the highest drug concentra tion. Migration assessed using a wound healing assay showed that this agent reduced the number of cells moving into the denuded area at concen trations 1 uM. Moreover, in the transwell assays, both cell migration and invasion were Discussion MET oncogene overexpression has been described in a variety of human cancers including prostate. Aber rant c Met activation has been shown Ixazomib Sigma to be strongly involved in prostate cancer aggressiveness and poorly clinical outcome.