Fig 1.
Mice lacking wild-type Hras develop more PanIN lesions and have reduced survival in oncogenic Kras-driven models of pancreatic cancer.
(A) Representative H & E stained sections (arrowheads: normal acinar cells), (B) quantification of % total normal acinar area remaining per field (at 4x magnification, 4–5 fields from 12 mice, bar: mean ± S.E.M.), (C) % of fields (at 4x magnification) with no normal acinar tissue (based on the data from B), and (D) % of lobules with the indicated highest grade lesion (Nml: normal, 1A: PanIN-1A, 1B: PanIN-1B, bar: mean ± S.E.M.) of pancreata isolated from Hras+/+ versus Hras-/- KC mice at 9 months of age. (E) Kaplan-Meier curve of Hras+/+ (n = 45) versus Hras-/- (n = 23) KPC mice. ns: not significant. *P<0.05. ***P<0.0001.
Fig 2.
Mice lacking wild-type Hras develop more oncogenic Kras-driven skin tumors.
(A) Representative photograph (white arrow) and (B) H & E stained section of facial papillomas that develop in KC mice. (C) Number of facial papillomas per Hras+/+ (n = 31) versus Hras-/- (n = 28) KC mice (bar: mean ± S.E.M.). (D) % of total Hras+/+ and Hras-/- KC mice (from C) that developed facial papillomas. (E) % facial papilloma-free survival of Hras+/+ (n = 45) versus Hras-/- (n = 23) KPC mice. (F) % vulvar papilloma-free survival of Hras+/+ (n = 19) versus Hras-/- (n = 11) female KPC mice. ***P<0.0001.
Fig 3.
Mice lacking wild-type Hras develop more ADM and PanIN lesions at early time points.
(A) Representative H & E stained section (arrowhead: PanIN lesion), (B) quantification of % total normal acinar area remaining per field (at 4x magnification, 5 fields from 10 mice, bar: mean ± S.E.M.), (C) % of fields with all normal acinar tissue (from B), and (D) % of lobules with the indicated highest grade lesion (Nml: normal, ADM: acinar-to-ductal metaplasia, 1A: PanIN-1A, bar: mean ± S.E.M.) of pancreata isolated from Hras+/+ versus Hras-/- KC mice at 8 weeks of age. (E) Representative H & E stained section of a pancreas from an 8-week old KC mouse (arrowhead: acinar-to-ductal metaplasia). (F) % of lobules with the indicated highest grade lesion (Nml: normal, ADM: acinar-to-ductal metaplasia, 1A: PanIN-1A bar: mean ± S.E.M.) of pancreata isolated from Hras+/+ versus Hras-/- KC mice at 4 weeks of age. (G) Representative pancreatic section from a 4-week old Hras+/+ versus Hras-/- KC mouse immunostained for cells (DAPI, blue) and markers of acinar (amylase, red) and ductal (CK-19, green) cells to highlight ADM lesions (co-staining, yellow). (H) Number of amylase+/CK-19+ positive (ADM) cells per field (at 4x magnification, (at 4x magnification, 5 fields from 10 mice) from pancreata isolated from Hras+/+ versus Hras-/- KC mice at 4 weeks of age (bar: mean ± S.E.M.). *P<0.05. ***P<0.0001.
Fig 4.
Wild-type Hras is activated in KPC cell lines and Hras+/+ and Hras-/- KPPC mice exhibit similar tumor burden and lifespan.
Immunoblot analysis of Hras and Hras-GTP in PDAC cell lines derived from (A) Hras-/- (cell lines 1–3) and Hras+/+ (cell lines 4–6) KPC mice or (B) PDAC cell lines 1 and 3 derived from Hras-/- KPC mice stably infected with a retrovirus encoding no transgene (vector, V) or wild-type Hras (H). Tubulin serves as a loading control. (C) Representative H&E stained sections from pancreata of Hras+/+ versus Hras-/- KPPC mice at 14 days of age. (4x magnification) (D) Quantification of the % normal acinar area remaining per field (at 4x magnification, 5 fields from 10 mice) from Hras+/+ versus Hras-/- KPPC mice (bar: mean ± S.E.M). (E) Kaplan-Meier survival curve of Hras+/+ (n = 20) versus Hras-/- (n = 11) KPPC mice. ns: not significant.