Twist expression promotes migration and invasion in hepatocellular carcinoma

Noriyuki Matsuo, Hidenori Shiraha, Tatsuya Fujikawa, Nobuyuki Takaoka, Naoki Ueda, Shigetomi Tanaka, Shinichi Nishina, Yutaka Nakanishi, Masayuki Uemura, Akinobu Takaki, Shinichiro Nakamura, Yoshiyuki Kobayashi, Kazuhiro Nouso, Takahito Yagi, Kazuhide Yamamoto

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Abstract

Background: Twist, 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).Methods: We 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.Results: We 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.Conclusion: Twist induces migratory effect on hepatocellular carcinoma by causing epithelial-mesenchymal transition.

Original languageEnglish
Article number240
JournalBMC Cancer
Volume9
DOIs
Publication statusPublished - Jul 18 2009

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Hepatocellular Carcinoma
Epithelial-Mesenchymal Transition
Focal Adhesions
Cell Movement
Cadherins
Cell Line
Small Interfering RNA
Twist-Related Protein 1
Western Blotting
Cell Migration Assays
Cell Proliferation
Vinculin
Hep G2 Cells
Vimentin
Wound Healing
Neoplasms
Immunohistochemistry
Staining and Labeling
Neoplasm Metastasis
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Oncology
  • Cancer Research
  • Genetics

Cite this

Matsuo, N., Shiraha, H., Fujikawa, T., Takaoka, N., Ueda, N., Tanaka, S., ... Yamamoto, K. (2009). Twist expression promotes migration and invasion in hepatocellular carcinoma. BMC Cancer, 9, [240]. https://doi.org/10.1186/1471-2407-9-240

Twist expression promotes migration and invasion in hepatocellular carcinoma. / Matsuo, Noriyuki; Shiraha, Hidenori; Fujikawa, Tatsuya; Takaoka, Nobuyuki; Ueda, Naoki; Tanaka, Shigetomi; Nishina, Shinichi; Nakanishi, Yutaka; Uemura, Masayuki; Takaki, Akinobu; Nakamura, Shinichiro; Kobayashi, Yoshiyuki; Nouso, Kazuhiro; Yagi, Takahito; Yamamoto, Kazuhide.

In: BMC Cancer, Vol. 9, 240, 18.07.2009.

Research output: Contribution to journalArticle

Matsuo, N, Shiraha, H, Fujikawa, T, Takaoka, N, Ueda, N, Tanaka, S, Nishina, S, Nakanishi, Y, Uemura, M, Takaki, A, Nakamura, S, Kobayashi, Y, Nouso, K, Yagi, T & Yamamoto, K 2009, 'Twist expression promotes migration and invasion in hepatocellular carcinoma', BMC Cancer, vol. 9, 240. https://doi.org/10.1186/1471-2407-9-240
Matsuo, Noriyuki ; Shiraha, Hidenori ; Fujikawa, Tatsuya ; Takaoka, Nobuyuki ; Ueda, Naoki ; Tanaka, Shigetomi ; Nishina, Shinichi ; Nakanishi, Yutaka ; Uemura, Masayuki ; Takaki, Akinobu ; Nakamura, Shinichiro ; Kobayashi, Yoshiyuki ; Nouso, Kazuhiro ; Yagi, Takahito ; Yamamoto, Kazuhide. / Twist expression promotes migration and invasion in hepatocellular carcinoma. In: BMC Cancer. 2009 ; Vol. 9.
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abstract = "Background: Twist, 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).Methods: We 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.Results: We 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.Conclusion: Twist induces migratory effect on hepatocellular carcinoma by causing epithelial-mesenchymal transition.",
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AU - Matsuo, Noriyuki

AU - Shiraha, Hidenori

AU - Fujikawa, Tatsuya

AU - Takaoka, Nobuyuki

AU - Ueda, Naoki

AU - Tanaka, Shigetomi

AU - Nishina, Shinichi

AU - Nakanishi, Yutaka

AU - Uemura, Masayuki

AU - Takaki, Akinobu

AU - Nakamura, Shinichiro

AU - Kobayashi, Yoshiyuki

AU - Nouso, Kazuhiro

AU - Yagi, Takahito

AU - Yamamoto, Kazuhide

PY - 2009/7/18

Y1 - 2009/7/18

N2 - Background: Twist, 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).Methods: We 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.Results: We 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.Conclusion: Twist induces migratory effect on hepatocellular carcinoma by causing epithelial-mesenchymal transition.

AB - Background: Twist, 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).Methods: We 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.Results: We 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.Conclusion: Twist induces migratory effect on hepatocellular carcinoma by causing epithelial-mesenchymal transition.

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