1. Ubiquitous Activation of Ras and Jak/Stat Pathways in Human HCC
Diego F. Calvisi, Sara Ladu, Alexis Gorden, Miriam Farina, Elizabeth A. Conner, Ju–Seog Lee, Valentina M. Factor, Snorri S. Thorgeirsson 
Gastroenterology 
Volume 130, Issue 4, Pages (April 2006)
DOI: /j.gastro
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

2. Figure 1
Ras signaling in human HCC. (A) Representative Western blot analysis of Ras inhibitors and proapoptotic molecules and (B) Ras downstream effectors. (C) Immunoprecipitation analysis showing that RASSF1A ability to bind both H-Ras and NORE1A is impaired in HCC and increased in nonneoplastic surrounding livers compared with normal livers. In contrast, RASSF2/K-ras complexes are markedly increased in HCC compared with nontumorous surrounding and normal livers. (D) Densitometric quantification of the band intensity for Ras inhibitors. Each bar represents integral optical density normalized to actin level and expressed as mean ± SE. 1, Normal liver (n = 6); 2, surrounding livers from HCC with better prognosis (n = 18); 3, surrounding livers from HCC with poor prognosis (n = 17); 4, HCC with better prognosis (n = 18); 5, HCC with poor prognosis (n = 17). a and b, P < .002 vs normal livers; c and d, P < 1.28E-06 vs respective surrounding livers, e, P = 4.028E-06 vs HCCb.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

3. Figure 2
Status of Ras inhibitors and Ras downstream effectors based on protein expression levels as detected by Western blot analysis in 35 human HCCs. Green square, down-regulation; yellow square, no difference; red square, up-regulation as compared with surrounding nonneoplastic livers. Difference in survival indicated by blue squares (<3 years, HCCp) and grey squares (>3 years, HCCb).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

4. Figure 3
Jak/Stat signaling in human HCC. (A) Representative Western blot analysis of Jak/Stat inhibitors, (B) tyrosine kinases, (C) Stat molecules, and (D) Jak/Stat target genes. (E and F) Densitometric quantification of the band intensity for Jak/STAT inhibitors (E) and tyrosine kinases (F). Each bar represents integral optical density normalized to actin level and expressed as mean ± SE. 1, Normal liver (n = 6); 2, surrounding livers from HCC with better prognosis (n = 18); 3, surrounding livers from HCC with poor prognosis (n = 17); 4, HCC with better prognosis (n = 18); 5, HCC with poor prognosis (n = 17). a and b, P < 2.96E-06 vs normal livers; c and d, P < 2.01E-05 vs respective surrounding livers, e = 2.70E-11 vs HCCb.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

5. Figure 4
Status of Jak/Stat inhibitors and members of Jak/Stat pathway based on protein expression levels as detected by Western blot analysis in 35 human HCCs. Green square, down-regulation; yellow square, no difference; red square, up-regulation as compared with surrounding nonneoplastic livers. Difference in survival indicated by blue squares (<3 years, HCCp) and grey squares (>3 years, HCCb).
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

6. Figure 5
Summary of promoter methylation of Ras and Jak/Stat inhibitors in a collection of normal and hepatoma cell lines (n = 20). Open squares represent absence and solid squares presence of promoter methylation.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

7. Figure 6
Combined treatment with Jak/Stat and Ras inhibitors induces massive apoptosis in hepatoma cell lines.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions

8. Figure 7
Zebularine treatment of hepatoma cell lines restores Ras (A) and Jak/Stat (B) inhibitor protein levels and induces cell death (C). Results obtained from the Hep3B cell line are shown. Experiments were repeated 3 times, and the data are expressed as mean ± SE.
Gastroenterology  , DOI: ( /j.gastro )
Copyright © 2006 American Gastroenterological Association Institute Terms and Conditions