Figure 1.
Delivery of pS/MAR-SMAD4 DNA Vectors Rescues the Tumorigenic Phenotype of SMAD4 Mutant Pancreatic Cancer Cell Lines
pS/MAR-luciferase (pS/MAR Luc) and pS/MAR-SMAD4-luciferase (pS/MAR SMAD4-Luc) DNA vectors were generated by introducing the transgene expression cassettes under the control of the ubiquitin C promoter (UbiC). (A) The expression of SMAD4 in modified Capan-1 was evaluated by real-time quantitative PCR (qPCR) and western blot in comparison to HEK293T cells, which constitutively express SMAD4. The impact of SMAD4 in the tumor growth was evaluated in vivo by injecting 5 × 105 Capan-1 cells expressing either the reporter gene luciferase or a combination of SMAD4 and luciferase orthotopically into the pancreas of NSG mice. (B) Capan-1 SMAD4-Luc cells generated significantly smaller tumors than did Capan-1 luciferase (n = 4 per group analyzed with one-way ANOVA followed by Tukey’s post hoc test for multiple comparisons, ∗p = 0.0141). (C) Histopathological analysis reveals that the luciferase-modified cells developed a tumor with identical morphology as those formed from the parental cell line, while the rescue of SMAD4 induces profound changes. (D) Capan-1 luciferase and Capan-1 SMAD4-Luc-derived tumors were assessed for histology by hematoxylin and eosin (H&E), proliferation (Ki67), and SMAD4 expression.