Figure 6
Cyr61 knockdown reduces mesenchymal transition and lung metastasis in a mouse model. (A) Total proteins were collected from MG63 cells stably expressing shRNAs directed against Cyr61. A vector-only control isshown (control sh). Western blot analysis was used to detect TWIST-1, N-cadherin, and E-cadherin. Actin was used as the loading control. (B-C) Total mRNA was collected from MG63 cells stably expressing shRNAs, and the expression levels of Cyr61, TWIST-1, N-cadherin and E-cadherin were detected using qRT-PCR. (D) MG63 cells stably expressing shRNA constructs were seeded as monolayers and were counted daily. Cells (1 × 104) were reseeded after each count, and the cell numbers were plotted. (E) The in vitro migration of MG63 cells stably expressing shRNA constructs was measured using the Transwell assay. (F-H) MG63 cells stably expressing control sh and Cyr61 sh2 cells were injected into the tail vein of CB17-SCID mice. After 4 wk, the mice were sacrificed, and the lung metastatic nodules were photographed and counted. The results are expressed as the mean ± SEM of triplicate samples. *P <0.05 compared with the control group.