Expression and Functional Significance of Twist1 in Hepatocellular Carcinoma: Its Role in Vasculogenic Mimicry

Abstract

The up-regulation and nuclear relocation of epithelial-mesenchymal transition (EMT) regulator Twist1 have been implicated in the tumor invasion and metastasis of human hepatocellular carcinoma (HCC). The term vasculogenic mimicry (VM) refers to the unique capability of aggressive tumor cells to mimic the pattern of embryonic vasculogenic networks. However, the relationship between Twist1 and VM formation is not clear. In this study, we explored HCC as a VM and EMT model in order to investigate the role of Twist1 in VM formation. We first examined the expression of Twist1 in human HCC samples and cell lines and found that Twist1 was frequently overexpressed in the nuclear relocation occurring in VM-positive HCCs (13/18 [72%]). Twist1 nuclear expression was likewise significantly associated with VM formation. Clinicopathological analysis revealed that both VM and Twist1 nuclear expressions present shorter survival durations than those without expression. We consistently demonstrated that an overexpression of Twist1 significantly enhanced cell motility, invasiveness, and VM formation in an HepG2 cell. Conversely, a knockdown of Twist1 by the short hairpin RNA approach remarkably reduced Bel7402 cell migration, invasion, and VM formation. Using chromatin immunoprecipitation, we also showed that Twist1 binds to the vascular endothelial (VE)-cadherin promoter and enhances its activity in a transactivation assay. Conclusion: The results of this study indicate that Twist1 induces HCC cell plasticity in VM cells more through the suppression of E-cadherin expression and the induction of VE-cadherin up-regulation than through the VM pattern in vivo and in a three-dimensional in vitro system. Our findings also demonstrate a novel cogitation in cancer stem-like cell differentiation and that related molecular pathways may be used as novel therapeutic targets for the inhibition of HCC angiogenesis and metastasis. (HEPATOLOGY 2010;51:545-556.)

Overexpression of HIF-1α in primary gallbladder carcinoma and its relation to vasculogenic mimicry and unfavourable prognosis

“…In addition, similar structures have been reported by several groups in other tumors using H&E staining and CD31/ CD34-PAS double staining (15,17,(41)(42)(43). VM was present in 18 of 71 (25.4%) cases by using H&E and CD31-PAS double staining, which is different from other tumors, such as ovarian carcinoma (36.4%) (13), breast cancer (7.9%) (12) and HCC (19%) (44). However, our results were similar to another report, such as LSCC (21.67%) (15).…”

Twist1 induces endothelial differentiation of tumour cells through the Jagged1-KLF4 axis

“…This observation is in drastic contrast to the traditional angiogenesis process contributed by surrounding endothelial progenitors that are triggered by cytokines and pro-angiogenic factors secreted from tumour-associated macrophages/ neutrophils and tumour-associated fibroblasts 2,3 . Induction of tumour-derived endothelial differentiation by Twist1 is different from the vasculogenic mimicry mechanism 33,34 , which does not activate endothelial markers and may represent an incomplete differentiation towards the endothelial lineage. This induction of endothelial differentiation may be critical for tumour progression and metastasis since knockdown of either Jagged1 or KLF4 significantly decreases the in vitro or in vivo metastatic activity induced by Twist1 ( Fig.…”

Tumor Blood Vessels and Vasculogenic Mimicry – Current Knowledge and Searching for New Cellular/Molecular Targets of Anti-Angiogenic Therapy

“…The latter agent is not the only adhesive molecule, but it may lead to induction of vascular cascade [23]. Earlier, Sun and co-workers [81] and Liu and co-workers [52] indicated a strong connection between EMT and VM. It is suggested that as a result of VM connection with EMT, tumor cells acquire features typical for stem cells phenotype (CSCs) with ability to invasion and migration [28].…”

“…An increase of VM density is connected with the increase of PGCCs amount. [10,57,92] mechanism of formation independent of VEGF, EMT contribution [52,54,57,81] interaction with ECM increase of MMPs activity and ECM remodeling [50,60] phenotype/ cell plasticity expression of stem cell multipotent phenotype (Sox-2, TWIST-1, CD133/CD44) with ability to ECs differentiation at the same time [46,68,74,84,85] tumor invasiveness and metastasis high tumor progresion and aggressiveness [50,63] prognosis for patient adverse prognostic factor, shorter survival of patient [8,50,51] Final classification of vessel-like structures needs usually more studies towards amplification of EGFR gene, which is often observed in tumor cells derived from epithelial tissue [26].…”