Twist expression promotes migration and invasion in hepatocellular carcinoma

Abstract

BackgroundTwist, a transcription factor of the basic helix-loop-helix class, is reported to regulate cancer metastasis. It is known to induce epithelial-mesenchymal transition (EMT). In this study, we evaluated the expression of twist and its effect on cell migration in hepatocellular carcinoma (HCC).MethodsWe examined twist expression using immunohistochemistry in 20 tissue samples of hepatocellular carcinoma, and assessed twist expression in HCC cell lines by RT-PCR and Western blot analysis. Ectopic twist expression was created by introducing a twist construct in the twist-negative HCC cell lines. Endogenous twist expression was blocked by twist siRNA in the twist-positive HCC cell lines. We studied EMT related markers, E-cadherin, Vimentin, and N-cadherin by Western blot analysis. Cell proliferation was measured by MTT assay, and cell migration was measured by in vitro wound healing assay. We used immunofluorescent vinculin staining to visualize focal adhesion.ResultsWe detected strong and intermediate twist expression in 7 of 20 tumor samples, and no significant twist expression was found in the tumor-free resection margins. In addition, we detected twist expression in HLE, HLF, and SK-Hep1 cells, but not in PLC/RPF/5, HepG2, and Huh7 cells. Ectopic twist-expressing cells demonstrated enhanced cell motility, but twist expression did not affect cell proliferation. Twist expression induced epithelial-mesenchymal transition together with related morphologic changes. Focal adhesion contact was reduced significantly in ectopic twist-expressing cells. Twist-siRNA-treated HLE, HLF, and SK-Hep1 cells demonstrated a reduction in cell migration by 50, 40 and 18%, respectively.ConclusionTwist induces migratory effect on hepatocellular carcinoma by causing epithelial-mesenchymal transition.

Silencing of Twist Expression by RNA Interference Suppresses Epithelial-mesenchymal Transition, Invasion, and Metastasis of Ovarian Cancer

“…After A2780 was transfected with the Twist-shRNA plasmid, the expressions of E-cadherin and N-cadherin increased and decreased, respectively. The results of the present study agree with the findings of previous studies on liver cancer and laryngocarcinoma (Matsuo et al, 2009). However, no conclusion was derived in terms of the effect of Twist on the proliferation ability of the tumor cells.…”

“…However, this locomotivity was decreased significantly after Twist-shRNA treatment. Thus, Twist promoted the transfer of liver cells through EMT (Matsuo et al, 2009). Therefore, we can determine that Twist gene silencing can inhibit the adhesion ability of the ovarian cancer cell line A2780.…”

“…However, no conclusion was derived in terms of the effect of Twist on the proliferation ability of the tumor cells. Some researchers found that Twist can promote tumor cell proliferation (Hasselblatt et al, 2009), whereas others found that it can promote the invasion of tumor cells and has no effect on cell multiplication (Matsuo et al, 2009). Hasselblatt et al (2009) found that Twist is expressed in choroidal carcinoma and can promote the proliferation and invasion of choroidal epithelial cells.…”

“…Twist is necessary in the transfer of adenocarcinoma cells from the breast to lungs (Yang et al, 2004). Noriyuk studied the liver cancer (Matsuo et al, 2009) and found no obvious expression of Twist on the slice edge without tumor. Twist promotes the transmission of liver cells through EMT, demonstrating the high motion ability of Twist.…”

Tumor initiation and progression in hepatocellular carcinoma: risk factors, classification, and therapeutic targets

“…Characteristics for EMT are a shift in gene expression with an upregulation of Twist [85,90] , Snail [91] , VE-cadherin [92] , Vimentin [93] , downregulation of E-cadherin [94] , hepatocyte transcriptional factor HNF4alpha and changes in the cytoskeleton. Important molecular pathway involved in EMT are Wnt/betacatenin signaling [95,96] and through growth factor signalling like PDGF [97] .…”