TY - JOUR
T1 - Interplay between oncogenic K-Ras and wild-type H-Ras in Caco2 cell transformation
AU - Ikonomou, Georgia
AU - Kostourou, Vassiliki
AU - Shirasawa, Senji
AU - Sasazuki, Takehiko
AU - Samiotaki, Martina
AU - Panayotou, George
N1 - Funding Information:
This work was supported by a grant from the Operational Program “Competitiveness”, General Secretariat of Research and Technology , Greece ( 03ED562 PENED ) to GP. We thank Dr. A. Pintzas for gift of materials and helpful comments on this work.
PY - 2012/9/18
Y1 - 2012/9/18
N2 - Mutation of RAS genes is one of the most common oncogenic alterations in cancer and acquisition of activating RAS mutations has been demonstrated to cause progression of colorectal adenoma to cancer. The aim of this study was to identify changes in the proteome of the intermediate-stage colorectal cancer cell line Caco2, induced by ectopic expression of two distinct RAS proteins, KRASV12 and HRASV12, in their mutated, constitutively active form. Using 2D-gel electrophoresis, followed by LC-MS/MS we identified almost 200 differentially expressed proteins in pair-wise comparisons of Caco2 vs Caco2-KRASV12 and Caco2 vs Caco2-HRASV12. Although many of the affected proteins were unique for each pair, there were also substantial similarities. Interestingly, transformation by the mutant KRASV12 gene resulted in elevated expression levels and activity of endogenous H-ras protein. Silencing the latter with a specific RNAi reversed several proteomic changes observed in KRASV12-transformed cells, suggesting that oncogenic K-ras partly exerts its effects through endogenous H-ras activation. Alterations in the expression of cytoskeletal and cell adhesion proteins, caused by HRAS siRNA treatment, correlated with a reduction in the invasive properties of Caco2-KRASV12 cells. Our data suggest a novel interplay between K-ras and H-ras, with possible implications for colorectal carcinogenesis.
AB - Mutation of RAS genes is one of the most common oncogenic alterations in cancer and acquisition of activating RAS mutations has been demonstrated to cause progression of colorectal adenoma to cancer. The aim of this study was to identify changes in the proteome of the intermediate-stage colorectal cancer cell line Caco2, induced by ectopic expression of two distinct RAS proteins, KRASV12 and HRASV12, in their mutated, constitutively active form. Using 2D-gel electrophoresis, followed by LC-MS/MS we identified almost 200 differentially expressed proteins in pair-wise comparisons of Caco2 vs Caco2-KRASV12 and Caco2 vs Caco2-HRASV12. Although many of the affected proteins were unique for each pair, there were also substantial similarities. Interestingly, transformation by the mutant KRASV12 gene resulted in elevated expression levels and activity of endogenous H-ras protein. Silencing the latter with a specific RNAi reversed several proteomic changes observed in KRASV12-transformed cells, suggesting that oncogenic K-ras partly exerts its effects through endogenous H-ras activation. Alterations in the expression of cytoskeletal and cell adhesion proteins, caused by HRAS siRNA treatment, correlated with a reduction in the invasive properties of Caco2-KRASV12 cells. Our data suggest a novel interplay between K-ras and H-ras, with possible implications for colorectal carcinogenesis.
UR - http://www.scopus.com/inward/record.url?scp=84865563871&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84865563871&partnerID=8YFLogxK
U2 - 10.1016/j.jprot.2012.06.038
DO - 10.1016/j.jprot.2012.06.038
M3 - Article
C2 - 22800643
AN - SCOPUS:84865563871
SN - 1874-3919
VL - 75
SP - 5356
EP - 5369
JO - Journal of Proteomics
JF - Journal of Proteomics
IS - 17
ER -
