Figure 4.

PLC/PKC/c-Src signaling pathway is involved in PGE₂-mediated migration and integrin upregulation in human chondrosarcoma cells. (A) JJ012 cells were incubated with PGE₂ for indicated time intervals, and p-PLCβ3, p-PKCα and p-c-Src expression was determined by Western blot analysis. (B&D) JJ012 cells were pretreated for 30 min with U73122, GF109203X or PP2. Then they were followed by stimulation with PGE₂, and in vitro migration and cell surface α2β1 integrin were measured with the Transwell and flow cytometry after 24 hr. (C) Cells were transfected with PKCα or c-Src mutant and PLCβ siRNA for 24 hr followed by stimulation with PGE₂, and in vitro migration measured with the Transwell after 24 hr. JJ012 cells were incubated with PGE₂ for indicated time intervals, and PKCα activity was determined by the PKCα kinase assay kit. Results are expressed as the mean ± S.E. *, p < 0.05 compared with control; #, p < 0.05 compared with PGE₂-treated group