Abstract
A large percentage of metastatic colorectal cancer (mCRC) patients presents metastasis at the time of diagnosis. In the last years, great efforts have been made in the treatment of these patients with the identification of different phenotypes playing a key role in the definition of new systemic therapies. Unsupervised hierarchical clustering analysis (HCA) was performed considering the clinicopathological characteristics of 51 mCRCs. Using immunohistochemistry on tissue microarrays, we assessed the expression of β-catenin, NHERF1, RASSF1A, TWIST1, HIF-1α proteins in tumors and paired liver metastases. We also analyzed RASSF1A methylation status on the samples of the same patients. HCA distinguished Group 1 and Group 2 characterized by different clinicopathological features. Group 1 was characterized by higher number of positive lymph nodes (p=0.0139), poorly differentiated grade (p<0.0001) and high extent of tumor spread (p=0.0053) showing a more aggressive phenotype compared to Group 2. In both Groups, we found a common "basal" condition with a higher level of nuclear TWIST1 (p<0.0001 and cytoplasmic β-catenin (p<0.0001) in tumors than in paired liver metastases. Furthermore, the Group 1 was also characterized by RASSF1A hypermethylation (p<0.0001) and nuclear HIF-1α overexpression (p=0.0354) in paired liver metastases than in tumors. In conclusion, HCA identifies mCRC patients with a more aggressive phenotype. Moroever, our results support the important contribution to the progression of the disease of RASSF1A methylation and the oncogenic role of HIF-1α in these patients. These evidences, should provide relevant information concerning the biology of this tumor and, as a consequence, potential new systemic therapeutic approaches.