The Erythropoietin/Erythropoietin Receptor Signaling Pathway Promotes Growth and Invasion Abilities in Human Renal Carcinoma Cells
Figure 3
Change of signaling molecules, proliferation, apoptosis, invasion ability and MMP-2 expression in 786-0 cells with knockdown EpoR.
(A) Signaling molecules were assayed in 786-0 expressing EpoR siRNA (EpoR siRNA) by western blot, using parental 786-0 cells (control) and 786-0 expressing negative siRNA (Neg siRNA) as controls. In 786-0 cells expressing EpoR siRNA, EpoR and p-EpoR decreased by >90%, p-STAT5 decreased by >60%, but p-Erk1/2 increased by >36%, as compared with those in the two controls. (B) Proliferation rate reduced significantly in 786-0 expressing EpoR siRNA by modified MTT method (experiments in triplicate; P<0.01), as compared with that at the same time point in the two controls. (C) Invasion ability decreased by about 55% in 786-0 expressing EpoR siRNA by transwell test (tests in triplicate; ★★★: P<0.001). (D) MMP-2 expression was inhibited by 33% in 786-0 expressing EpoR siRNA by gelatin zymography, and MMP-9 expression was very low in the 3 groups of 786-0 cells. (E) Early apoptotic cells (annexin V-PE+/7-AAD-) increased by about 4 times in 786-0 expressing EpoR siRNA, as compared to those of 786-0 expressing negative siRNA by flow cytometry (experiments in triplicate; ★★★: P<0.001).