SMURF1 Amplification Promotes Invasiveness in Pancreatic Cancer
Figure 4
Analysis of SMURF1 knockdown in other pancreatic cancer lines.
(A) Western blot analysis of endogenous SMURF1 protein levels in a panel of pancreatic cancer cell lines. Note that SMURF1 is highly expressed only in the 7q22.1-amplified AsPC-1 cell line. Two different exposures of the SMURF1 blot are shown; GAPDH serves as a loading control. (B) Western blot verification of SMURF1 knockdown (by SMURF1-targeting siRNA pool, compared to non-targeting control siRNA pool) in BxPC-3 and Hs700T cells. (C) Cell proliferation/viability assayed (by WST-1) in BxPC-3 cells following SMURF1 siRNA-mediated knockdown (compared to non-targeting control). Assay done in triplicate (mean +/− 1SD shown); *, P<0.05 (Student's t-test). (D) Cell proliferation/viability assayed in Hs700T cells, as above. (E) Cell invasion assayed (by Boyden chamber) in BxPC-3 cells following SMURF1 knockdown (compared to non-targeting control). Assay done in triplicate (mean +/− 1SD shown). Note, the reduced proliferation observed with SMURF1 knockdown in Hs700T cells precluded an assay of cell invasion (where invaded cell numbers at 72 hrs are influenced by doubling times).