The T-box transcription factor Brachyury regulates epithelial-mesenchymal transition in association with cancer stem-like cells in adenoid cystic carcinoma cells.

BMC Cancer 2012 Aug 29;12:377. Epub 2012 Aug 29.

Division of Maxillofacial Diagnostic and Surgical Sciences, Department of Oral and Maxillofacial Surgery, Graduate School of Dental Science, Kyushu University, 3-1-1 Maidashi, Fukuoka 812-8582, Japan.

Background: The high frequencies of recurrence and distant metastasis of adenoid cystic carcinoma (AdCC) emphasize the need to better understand the biological factors associated with these outcomes. To analyze the mechanisms of AdCC metastasis, we established the green fluorescence protein (GFP)-transfected subline ACCS-GFP from the AdCC parental cell line and the metastatic ACCS-M GFP line from an in vivo metastasis model.

Methods: Using these cell lines, we investigated the involvement of the epithelial-mesenchymal transition (EMT) and cancer stem cell (CSCs) in AdCC metastasis by real-time RT-PCR for EMT related genes and stem cell markers. Characteristics of CSCs were also analyzed by sphere-forming ability and tumorigenicity. Short hairpin RNA (shRNA) silencing of target gene was also performed.

Results: ACCS-M GFP demonstrated characteristics of EMT and additionally displayed sphere-forming ability and high expression of EMT-related genes (Snail, Twist1, Twist2, Slug, zinc finger E-box binding homeobox 1 and 2 [Zeb1 and Zeb2], glycogen synthase kinase 3 beta [Gsk3β and transforming growth factor beta 2 [Tgf-β2]), stem cell markers (Nodal, Lefty, Oct-4, Pax6, Rex1, and Nanog), and differentiation markers (sex determining region Y [Sox2], Brachyury, and alpha fetoprotein [Afp]). These observations suggest that ACCS-M GFP shows the characteristics of CSCs and CSCs may be involved in the EMT of AdCC. Surprisingly, shRNA silencing of the T-box transcription factor Brachyury (also a differentiation marker) resulted in downregulation of the EMT and stem cell markers. In addition, sphere-forming ability, EMT characteristics, and tumorigenicity were simultaneously lost. Brachyury expression in clinical samples of AdCC was extremely high and closely related to EMT. This finding suggests that regulation of EMT by Brachyury in clinical AdCC may parallel that observed in vitro in this study.

Conclusions: The use of a single cell line is a limitation of this study. However, parallel data from in vitro and clinical samples suggest the possibility that EMT is directly linked to CSCs and that Brachyury is a regulator of EMT and CSCs.

Download full-text PDF

Source
http://dx.doi.org/10.1186/1471-2407-12-377DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3492149PMC
August 2012
4 Reads

Publication Analysis

Top Keywords

stem cell
16
accs-m gfp
12
sphere-forming ability
12
cell markers
12
emt
10
adenoid cystic
8
cystic carcinoma
8
adcc metastasis
8
epithelial-mesenchymal transition
8
shrna silencing
8
clinical samples
8
characteristics cscs
8
t-box transcription
8
factor brachyury
8
transcription factor
8
cell
7
adcc
7
brachyury
6
cscs
6
silencing target
4

References

(Supplied by CrossRef)

AD Rapidis et al.
Oral Oncol 2005

FL Ampil et al.
J Oral Maxillofac Surg 1987

K Ishii et al.
Int J Oncol 2011

D Tarin et al.
Cancer Res 2005

EW Thompson et al.
Cancer Res 2005

S Grunert et al.
Nat Rev Mol Cell Biol 2003

MA Nieto et al.
Nat Rev Mol Cell Biol 2002

V Bolos et al.
J Cell Sci 2003

MA Huber et al.
Curr Opin Cell Biol 2005

J Yang et al.
Cell 2004

A Cano et al.
Nat Cell Biol 2000

Similar Publications