Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=26165819
&cmd=llinks): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests
in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 215
Hepatic stellate cells promote upregulation of epithelial cell adhesion molecule
and epithelial-mesenchymal transition in hepatic cancer cells
#MMPMID26165819
Nagahara T
; Shiraha H
; Sawahara H
; Uchida D
; Takeuchi Y
; Iwamuro M
; Kataoka J
; Horiguchi S
; Kuwaki T
; Onishi H
; Nakamura S
; Takaki A
; Nouso K
; Yamamoto K
Oncol Rep
2015[Sep]; 34
(3
): 1169-77
PMID26165819
show ga
Microenvironment plays an important role in epithelial-mesenchymal transition
(EMT) and stemness of cells in hepatocellular carcinoma (HCC). Epithelial cell
adhesion molecule (EpCAM) is known as a tumor stemness marker of HCC. To
investigate the relationship between microenvironment and stemness, we performed
an in vitro co-culture assay. Four HCC cell lines (HepG2, Hep3B, HuH-7 and
PLC/PRF/5) were co-cultured with the TWNT-1 immortalized hepatic stellate cells
(HSCs), which create a microenvironment with HCC. Cell proliferation ability was
analyzed by flow cytometry (FCM) and
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, while
migration ability was assessed by a wound healing assay. Expression of EpCAM was
analyzed by immunoblotting and FCM. HCC cell lines were co-cultured with TWNT-1
treated with small interfering RNA (siRNA) for TGF-? and HB-EGF; we then analyzed
proliferation, migration ability and protein expression using the methods
described above. Proliferation ability was unchanged in HCC cell lines
co-cultured with TWNT-1. Migration ability was increased in HCC cell lines
(HepG2, Hep3B, HuH-7 and PLC/PRF/5) directly (216.2±67.0, 61.0±22.0, 124.0±66.2
and 51.5±40.3%) and indirectly (102.5±22.0, 84.6±30.9, 86.1±25.7 and 73.9±29.7%)
co-cultured with TWNT-1 compared with the HCC uni-culture. Immunoblot analysis
revealed increased EpCAM expression in the HCC cell lines co-cultured with
TWNT-1. Flow cytometry revealed that the population of E-cadherin-/N-cadherin+
and EpCAM-positive cells increased and accordingly, EMT and stemness in the HCC
cell line were activated. These results were similar in the directly and
indirectly co-cultured samples, indicating that humoral factors were at play.
Conversely, HCC cell lines co-cultured with siRNA?treated TWNT-1 showed decreased
migration ability, a decreased population of EpCAM-positive and
E-cadherin-/N-cadherin+ cells. Taken together, humoral factors secreted from
TWNT-1 promote upregulation of EpCAM and EMT in hepatic cancer cells.
free
free
free
