Journal Article

Cripto-1 contributes to stemness in hepatocellular carcinoma by stabilizing Dishevelled-3 and activating Wnt/β-catenin pathway

Chan, Kristy Kwan-Shuen, Wong, Chun-Ming, Lo, Regina Cheuk-Lam, Leung, Carmen Oi-Ning, Ng, Irene Oi-Lin, Lee, Terence Kin-Wah, Ho, Daniel Wai-Hung

2018

Overview

Identification and characterization of functional molecular targets conferring stemness properties in hepatocellular carcinoma (HCC) offers crucial insights to overcome the major hurdles of tumor recurrence, metastasis and chemoresistance in clinical management. In the current study, we investigated the significance of Cripto-1 in contributing to HCC stemness. Cripto-1 was upregulated in the sorafenib-resistant clones derived from HCC cell lines and patient-derived xenograft that we previously developed, suggesting an association between Cripto-1 and stemness. By in vitro experiments, Cripto-1 fostered cell proliferation, migration, and invasion. It also enhanced self-renewal ability and conferred chemoresistance of HCC cells. Consistently, silencing of Cripto-1 suppressed in vivo tumorigenicity on serial transplantation. On the downstream signaling mechanism, expression of major components of Wnt/β-catenin pathway β - catenin, AXIN2, and C-MYC, accompanied by β-catenin activity was reduced upon Cripto-1 knockdown. The suppressive effects on stemness properties with Cripto-1 knockdown in vitro and in vivo were partially rescued by forced expression of constitutively active β - catenin. Further elucidation revealed the binding of Cripto-1 to Frizzled-7 (FZD7), low-density lipoprotein receptor-related protein 6 (LRP6) and Dishevelled-3 (DVL3) of the Wnt/β - catenin pathway and stabilized DVL3 protein. Analyses with clinical samples validated Cripto-1 overexpression in HCC tissues, as well as a positive correlation between Cripto-1 and AXIN2 expressions. High Cripto-1 level in tumor was associated with poorer disease-free survival of HCC patients. Taken together, Cripto-1 binds to FZD7/LRP6 and DVL3, stabilizes DVL3 expression and activates the Wnt/β-catenin signaling cascade to confer stemness in HCC. Our study findings substantiated the role of Cripto-1 in determining stemness phenotypes of HCC and mechanistically in modulating the Wnt/β-catenin signaling cascade, one of the most frequently deregulated pathways in liver cancer.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 13/106 / 13/31 / 13/51 / 631/67/395 / 631/67/71 / 82/80 / 96/100 / 96/2 / Animals / Apoptosis / Apoptosis - drug effects / Axin Protein - genetics / Axin Protein - metabolism / beta Catenin - metabolism / Biochemistry / Biomedical and Life Sciences / c-Myc protein / Carcinoma, Hepatocellular - metabolism / Carcinoma, Hepatocellular - pathology / Cell Biology / Cell Cycle Analysis / Cell Line, Tumor / Cell migration / Cell proliferation / Cell self-renewal / Chemoresistance / Dishevelled protein / Dishevelled Proteins - chemistry / Dishevelled Proteins - metabolism / Drug Resistance, Neoplasm - drug effects / Frizzled protein / Frizzled Receptors - chemistry / Frizzled Receptors - metabolism / GPI-Linked Proteins - antagonists & inhibitors / GPI-Linked Proteins - genetics / GPI-Linked Proteins - metabolism / Hepatocellular carcinoma / Humans / Intercellular Signaling Peptides and Proteins - genetics / Intercellular Signaling Peptides and Proteins - metabolism / Life Sciences / Liver cancer / Liver Neoplasms - metabolism / Liver Neoplasms - pathology / Low Density Lipoprotein Receptor-Related Protein-6 - chemistry / Low Density Lipoprotein Receptor-Related Protein-6 - metabolism / Metastases / Mice / Mice, Inbred NOD / Mice, SCID / Myc protein / Neoplasm Proteins - antagonists & inhibitors / Neoplasm Proteins - genetics / Neoplasm Proteins - metabolism / Phenotypes / Protein Binding / Receptor density / RNA Interference / RNA, Small Interfering - metabolism / Signal transduction / Sorafenib - pharmacology / Stem Cells / Transplantation / Tumorigenicity / Wnt protein / Wnt Signaling Pathway / Xenografts / β-Catenin