Oppositely, the wounds were still

widely open at 24 hours after exposure to PTL at indicated concentrations. The results indicated that PTL treatment could inhibit migration of pancreatic cancer cell. Figure 3 PTL suppressed BxPC-3 migration. The wound gap of cells was scratched by a micropipette tip. Cells were incubated in the presence of PTL. 24 hours BMN-673 later the wound gap of BxPC-3 in the control group was nearly closed. On the contrary, after exposure to PTL at 7.5 μM, the speed of wound closure was much slower and the wound was still widely open at twenty-four hours. PTL inhibited BxPC-3 cell invasion The effect of PTL on BxPC-3 cell invasion was detected by a reconsitituted Matrigel membrane. The number of cells that passed through the filter and into the lower chamber was counted and compared. As a result, PTL at different concentrations obviously inhibited invasive ability of pancreatic cancer cell. The cell numbers of 7.5 μM and 15 μM PTL groups were (94 ± 7)/HPF and (58 ± 8)/HPF respectively, which were less than (146 ± 10)/HPF of control group (P < 0.05) (Fig. 4). Figure

4 PTL inhibited BxPC-3 cells invasion. Cells were fixed, stained and counted at 48 hours. The invasion cell numbers in PTL-treated groups were significantly less than the control group (P < 0.05), which indicated that PTL suppressed cell invasion dose-dependently. PTL downregulated Bcl-2 and upregulated Bax expression. No change was found on Bad The underlying mechanism of PTL was also explored in the study. The activation of several apoptosis-related proteins Erismodegib concentration may contribute to PTL-induced apoptosis.

In Bcl-2 family members, the expression of Bcl-2, Bax and Bad after PTL treatment for 48 hours were detected by Western blotting (Fig. 5A). PTL obviously decreased the protein expression of antiapoptotic Bcl-2 and increased the protein expression of proapoptotic Bax in the BxPC-3 cells after being treated with indicated concentrations. No change was found on Bad. Therefore, the susceptibility to PTL-induced apoptosis might be attributable to the imbalance of Bcl-2/Bax (Fig. 5B). Figure 5 Apoptosis-related protein expression Monoiodotyrosine after PTL treatment for 48 hours. (A) PTL decreased Bcl-2 expression and induced Bax expression. No obvious change was observed on Bad; (B) Bcl-2/Bax ratio was decreased significantly with the increasing concentration of PTL; (C) Activation of caspase-9 and caspase-3 after PTL treatment. Effect of PTL at various concentrations on caspase-9 and caspase-3 expression Data have showed many anticancer agents are capable of initiating the caspase activation and inducing apoptosis [14]. Hence the caspase cascade in PTL-induced effect was also analyzed. After PTL treatment at indicated concentrations for 48 hours, Caspase-9 and Pro-caspase-3 expressions of BxPC-3 cell were explored (Fig. 5C). The dose-dependent proteolytic cleavage of caspase-9 was detected.