β-catenin interaction with NHERF1 and RASSF1A methylation in metastatic colorectal cancer patients

Abstract

There is an increasing need to identify new biomarkers in colorectal cancer (CRC) to further characterize this malignancy. β-catenin plays a central role in the Wnt signaling pathway. It also binds Na+/H+ exchanger regulating factor 1 (NHERF1) and interacts with the RAS-association domain family 1, isoform A (RASSF1A), but the mechanisms of this possible crosstalk are still not fully understood. In this study, we analyzed for the first time the different subcellular expression of β-catenin, NHERF1, and RASSF1A and their relationships with RASSF1A methylation in the progression of CRC. We assessed immunohistochemical expression and RASSF1A methylation in 51 patients with stage IV colorectal cancer. Biomarker expression analysis was carried out considering the tumor-adjacent normal tissue, the primary tumor, and the paired liver metastases. Regarding the tumor compartment, it was found that cytoplasmic β-catenin expression was positively correlated to membranous (r = 0.3002, p = 0.0323) and nuclear NHERF1 (r = 0.293, p = 0.0368). In the liver metastases, instead, we found a positive correlation of cytoplasmic and nuclear β-catenin expression with RASSF1A methylation (r = 0.4019, p = 0.0068 and r = 0.3194, p = 0.0345, respectively).In conclusion, our results showed that β-catenin was the crucial protagonist in metastatic CRC through different effector proteins involved in this developing process. In tumor tissues, β-catenin was predominantly associated with NHERF1 in a dynamic context, while interestingly in liver metastases, we noted an increase of its oncogenic function through RASSF1A inactivation.

Hierarchical clustering analysis identifies metastatic colorectal cancers patients with more aggressive phenotype

“…In Group 1, we observed a higher level of RASSF1A methylation in paired liver metastases than in primary tumors. In accordance with previous data, changes in DNA methylation can play a key role in the metastatic process [ 19 ]. In particular, a crucial role for RASSF1A in CRC progression has been demonstrated [ 9 , 20 ].…”

“…For negative control, the primary antibody was omitted and replaced by PBS 1X pH 7.6. The accuracy, reliability and reproducibility of these antibodies (β-catenin, NHERF1, TWIST1, HIF-1α and RASSF1A) have been validated in previous studies already published [ 19 , 30 , 39 , 40 ].…”

Independent Negative Prognostic Role of TCF1 Expression within the Wnt/β-Catenin Signaling Pathway in Primary Breast Cancer Patients

“…The immunostaining of NHERF1 and β-catenin was assessed, as previously described [12,15]. NHERF1 immunostaining was predominantly cytoplasmic, and, in some cases, intense nuclear staining was also observed.…”

“…A previous study, which we carried out demonstrated the prognostic significance of nuclear NHERF1 expression (nNHERF1) in a cohort of patients with primary BC [13]. It is worth mentioning that NHERF1 plays a significant role during the regulation of several oncogenic signaling pathways, including Wnt/β-catenin [14,15]. Wnt/β-catenin pathway is an evolutionarily conserved pathway that controls cell proliferation, differentiation, and the maintenance of stem cells [16].…”

NHERF1 Between Promises and Hopes: Overview on Cancer and Prospective Openings

“…Further, new evidences in favor of involvement of NHERF1 in Wnt signaling pathway and consequently in CRC development have been collected. A positive correlation among RAS-association domain family 1 methylation, isoform A (RASSF1A) and nuclear β-catenin has been found too, noting that Wnt pathway activation is mediated by RASSF1A, a putative tumor suppressor RAS effector, whose epigenetic silencing by promoter methylation has been reported during cancer progression [51] , [52] . These observations support the NHERF1 limelight in the cancer development and its trigger role, given by its intrinsic nature, that makes it a highly versatile molecule.…”