Downregulation of DOCK1 sensitizes bladder cancer cells to cisplatin through preventing epithelial–mesenchymal transition

Figures & data

Figure 1 Different cisplatin chemosensitivity in bladder cancer cells.

Notes: (A) Protein expression of DOCK1 in bladder cancer cells. Three bladder cancer cell lines, including UM-UC-3, J82, and B87, were incubated with cisplatin for 48 hours. Cell viability and DNA synthesis were measured using CCK-8 method (B) and EdU incorporation assay (C), respectively. Magnification: ×200. ***P<0.001.
Abbreviations: DOCK1, dedicator of cytokinesis 1; h, hours.

Figure 2 Downregulation of DOCK1 sensitized bladder cancer cells to cisplatin.

Notes: Three bladder cancer cell lines, including UM-UC-3, J82, and B87, were transfected with DOCK1 siRNA and incubated with cisplatin for 48 hours. The CCK-8 assay was performed to determine cell viability in UM-UC-3 (A), J82 (B), and B87 (C) cells. (D) Protein expression of DOCK1 in bladder cancer cells transfected with DOCK1 siRNA. ***P<0.001.
Abbreviations: DOCK1, dedicator of cytokinesis 1; siRNA, small interfering RNA.

Figure 3 Cisplatin treatment induced EMT in bladder cancer cells.

Notes: Three bladder cancer cell lines, including UM-UC-3, J82, and B87, were incubated with cisplatin for 48 hours. Immunofluorescence (A) was used to detect the expression of EMT-related markers, including E-cadherin and vimentin. Magnification: ×200. (B) Protein expression of Twist in bladder cancer cells transfected with Twist siRNA. (C) CCK-8 assay was performed to determine the response of Twist siRNA-transfected cells to cisplatin. ***P<0.001.
Abbreviations: EMT, epithelial–mesenchymal transition; siRNA, small interfering RNA.

Figure 4 DOCK1 regulated EMT in bladder cancer cells.

Notes: Western blot analysis of E-cadherin and vimentin expression in UM-UC-3, J82, and B87 before (A) and after transfection with DOCK1 siRNA (B). ***P<0.001.
Abbreviations: DOCK1, dedicator of cytokinesis 1; EMT, epithelial–mesenchymal transition; siRNA, small interfering RNA.

Figure 5 Cisplatin-induced EMT was dependent on DOCK1.

Notes: (A) Western blot analysis of DOCK1 expression in bladder cancer cells exposed to cisplatin. (B) After transfection with DOCK1 siRNA, the expression of DOCK1, E-cadherin, and vimentin was determined in the presence of cisplatin. ***P<0.001.
Abbreviations: EMT, epithelial–mesenchymal transition; DOCK1, dedicator of cytokinesis 1; siRNA, small interfering RNA.

Figure 6 DOCK1 knockdown sensitized bladder cancer cells to cisplatin via EMT.

Notes: After transfection with Twist siRNA alone or in combination with DOCK1 siRNA, CCK-8 assay was performed to determine the cell viability of UM-UC-3 (A), B87 (B), and J82 (C) cells. (D) Western blotting was used to measure the protein expression of DOCK1, Twist, E-cadherin, and vimentin in bladder cancer cells. **P<0.01; ***P<0.001.
Abbreviations: DOCK1, dedicator of cytokinesis 1; EMT, epithelial–mesenchymal transition; siRNA, small interfering RNA.