Original Article
Transforming growth factor-b-induced plasticity causes a migratory
stemness phenotype in hepatocellular carcinoma
Q8
Q7
Andrea Malfettone
a
, Jitka Soukupova
a
, Esther Bertran
a
, Eva Crosas-Molist
a, 1
,
Raquel Lastra
b
, Joan Fernando
a
, Petra Koudelkova
c
, Bhavna Rani
d
,
Angels Fabra
a
,
Teresa Serrano
e
, Emilio Ramos
b
, Wolfgang Mikulits
c
, Gianluigi Giannelli
f
,
Isabel Fabregat
a, g, *
a
Bellvitge Biomedical Research Institute (IDIBELL), L'Hospitalet, Barcelona, Spain
b
Department of Surgery, Liver Transplant Unit, University Hospital of Bellvitge, Barcelona, Spain
c
Department of Medicine I, Division: Institute of Cancer Research, Comprehensive Cancer Center Vienna, Medical University of Vienna, Vienna, Austria
d
Department of Biomedical Sciences and Human Oncology, University of Bari, Medical School, Bari, Italy
e
Pathological Anatomy Service, University Hospital of Bellvitge, Barcelona, Spain
f
National Institute of Gastroenterology IRCCS “S. De Bellis” Castellana Grotte Bari, Italy
g
Department of Physiological Sciences, School of Medicine, University of Barcelona, Barcelona, Spain
article info
Article history:
Received 5 December 2016
Received in revised form
24 January 2017
Accepted 25 January 2017
Keywords:
CD44
EMT
HCC
TGF-beta
Stem
abstract
As part of its potential pro-tumorigenic actions, Transforming Growth Factor-(TGF)-b induces epithelial
emesenchymal transition (EMT) in hepatocellular carcinoma (HCC) cells. Whether EMT induces changes
in tumor cell plasticity has not been fully explored yet. Here, we analyze the effects of TGF-b on the EMT
and stem-related properties of HCC cells and the potential correlation among those processes. The
translational aim of the study was to propose a TGF-b/EMT/stem gene signature that would help in
recognizing HCC patients as good candidates for anti-TGF-b therapy. Results indicate that when TGF-b
induces EMT in HCC cells, a switch in the expression of stem genes is observed and their stemness
potential and migratory/invasive capacity are enhanced. However, TGF-b may induce a partial EMT in
some epithelial HCC cells, increasing the expression of mesenchymal genes and CD44, but maintaining
epithelial gene expression. Epithelial cells show higher stemness potential than the mesenchymal ones,
but respond to TGF-b increasing their migratory and invasive capacity. In HCC patient samples, TGFB1
expression most frequently correlates with a partial EMT, increase in mesenchymal genes and CD44
expression, as well as maintenance or over-expression of epithelial-related genes.
© 2017 Elsevier B.V. All rights reserved.
Introduction
The Transforming Growth Factor (TGF)-b inhibits growth and
induces apoptosis in hepatocytes, thus representing a potent tu-
mor suppressor in the liver [1]. However, once cells overcome
TGF-b-induced anti-oncogenic responses, this cytokine contrib-
utes to tumor progression and metastasis through the induction
of epithelialemesenchymal transition (EMT), which increases cell
migration and invasion [2]. Liver tumors expressing late TGF-b-
responsive genes (anti-apoptotic and EMT-related) display a
higher invasive phenotype and increased tumor recurrence when
compared to those that show an early TGF-b signature (sup-
pressor genes) [3]. In line with this theory, we showed that
overactivation of the TGF-b pathway in hepatocellular carcinoma
(HCC) cells confers them a mesenchymal-like phenotype and
increased migratory properties [4]. For all these reasons, target-
ing TGF-b has recently emerged as a promising tool in fighting
liver cancer [5]. However, the design of new biomarkers that
reveal the pro-tumorigenic role of TGF-b in the patients will be
essential for proper stratification of those benefiting from anti-
TGF-b treatment.
Abbreviations: CI, Cell Index; CK-18, cytokeratin-18; CSCs, cancer stem cells; EMT,
epithelialemesenchymal transition; FBS, fetal bovine serum; HCC, hepatocellular
carcinoma; mRNA, messenger RNA; qRT-PCR, quantitative reverse-transcriptase
polymerase chain reaction; SEM, standard error of mean; shRNA, short hairpin
RNA; TGF, Transforming Growth Factor; TbRI, TGF-b Receptor I; TbT-Hep3B, TGF-b-
treated Hep3B; TbT-PLC, TGF-b-treated PLC/PRF/5.
* Corresponding author. Bellvitge Biomedical Research Institute (IDIBELL), Gran
via de l'Hospitalet, 199, L'Hospitalet, 08908 Barcelona, Spain. Fax: þ34 932607426.
E-mail address: ifabregat@idibell.cat (I. Fabregat).
1
Current address: Tumour Plasticity Laboratory, Randall Division of Cell and
Molecular Biophysics, New Hunt's House, Guy's Campus, King's College London.
Contents lists available at ScienceDirect
Cancer Letters
journal homepage: www.elsevier.com/locate/canlet
http://dx.doi.org/10.1016/j.canlet.2017.01.037
0304-3835/© 2017 Elsevier B.V. All rights reserved.
Cancer Letters xxx (2017) 1e12
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Please cite this article in press as: A. Malfettone, et al., Transforming growth factor-b-induced plasticity causes a migratory stemness phenotype
in hepatocellular carcinoma, Cancer Letters (2017), http://dx.doi.org/10.1016/j.canlet.2017.01.037