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Open Access Highly Accessed Research article

Conformational changes and translocation of tissue-transglutaminase to the plasma membranes: role in cancer cell migration

Ambrish Kumar1, Jianjun Hu2, Holly A LaVoie3, Kenneth B Walsh4, Donald J DiPette5 and Ugra S Singh1*

Author Affiliations

1 Department of Pathology, Microbiology and Immunology, School of Medicine, University of South Carolina, Columbia, SC 29209, USA

2 Department of Computer Science and Engineering, University of South Carolina, Columbia, SC, USA

3 Department of Cell Biology and Anatomy, School of Medicine, University of South Carolina, Columbia, SC, USA

4 Department of Pharmacology, Physiology and Neuroscience, School of Medicine, University of South Carolina, Columbia, SC, USA

5 Department of Internal Medicine, School of Medicine, University of South Carolina, Columbia, SC, USA

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BMC Cancer 2014, 14:256  doi:10.1186/1471-2407-14-256

Published: 11 April 2014

Abstract

Background

Tissue-transglutaminase (TG2), a dual function G-protein, plays key roles in cell differentiation and migration. In our previous studies we reported the mechanism of TG2-induced cell differentiation. In present study, we explored the mechanism of how TG2 may be involved in cell migration.

Methods

To study the mechanism of TG2-mediated cell migration, we used neuroblastoma cells (SH-SY5Y) which do not express TG2, neuroblastoma cells expressing exogenous TG2 (SHYTG2), and pancreatic cancer cells which express high levels of endogenous TG2. Resveratrol, a natural compound previously shown to inhibit neuroblastoma and pancreatic cancer in the animal models, was utilized to investigate the role of TG2 in cancer cell migration. Immunofluorescence assays were employed to detect expression and intracellular localization of TG2, and calcium levels in the migrating cells. Native gel electrophoresis was performed to analyze resveratrol-induced cellular distribution and conformational states of TG2 in migrating cells. Data are presented as the mean and standard deviation of at least 3 independent experiments. Comparisons were made among groups using one-way ANOVA followed by Tukey-Kramer ad hoc test.

Results

TG2 containing cells (SHYTG2 and pancreatic cancer cells) exhibit increased cell migration and invasion in collagen-coated and matrigel-coated transwell plate assays, respectively. Resveratrol (1 μM-10 μM) prevented migration of TG2-expressing cells. During the course of migration, resveratrol increased the immunoreactivity of TG2 without affecting the total TG2 protein level in migrating cells. In these cells, resveratrol increased calcium levels, and depletion of intracellular calcium by a calcium chelator, BAPTA, attenuated resveratrol-enhanced TG2 immunoreactivity. In native-polyacrylamide gels, we detected an additional TG2 protein band with slower migration in total cell lysates of resveratrol treated cells. This TG2 form is non-phosphorylated, exclusively present in plasma membrane fractions and sensitive to intracellular Ca2+ concentration suggesting a calcium requirement in TG2-regulated cell migration.

Conclusions

Taken together, we conclude that resveratrol induces conformational changes in TG2, and that Ca2+-mediated TG2 association with the plasma membrane is responsible for the inhibitory effects of resveratrol on cell migration.

Keywords:
Cell migration; Invasion; Neuroblastoma; Resveratrol; Tissue-transglutaminase