The Expression of CD90/Thy-1 in Hepatocellular Carcinoma: An In Vivo and In Vitro Study
et al. (2013) The Expression of CD90/Thy-1 in Hepatocellular Carcinoma: An In
Vivo and In Vitro Study. PLoS ONE 8(10): e76830. doi:10.1371/journal.pone.0076830
The Expression of CD90/Thy-1 in Hepatocellular Carcinoma: An In Vivo and In Vitro Study
Caecilia Hapsari Ceriapuri Sukowati 0
Beatrice Anfuso 0
Giuliano Torre 0
Paola Francalanci 0
Lory 0
Saveria Croc 0
Claudio Tiribelli 0
Antonio Paolo Beltrami, University of Udine, Italy
0 1 Centro Studi Fegato, Fondazione Italiana Fegato , Trieste , Italy , 2 Hepatology Unit, Gastroenterology and Nutrition, Department of Surgery and Transplantation, Ospedale Pediatrico Bambino Gesu , Rome , Italy , 3 Department of Laboratories, Ospedale Pediatrico Bambino Gesu , Rome , Italy , 4 Department of Medical Sciences, University of Trieste , Trieste , Italy
Although the CD90 (Thy-1) was proposed as biomarker of several tumors and cancer stem cells, the involvement of this molecule in the progression of hepatocellular carcinoma (HCC) and other less frequent hepatic neoplasms is still undefined. The distribution of CD90 was investigated both in in vivo (human tissues samples) and in vitro (human HCC cell line JHH-6). A total of 67 liver tumors were analyzed: 51 HCC, 6 cholangiocarcinoma and 10 hepatoblastoma. In all cases, paired tissue sample of both the tumor and cirrhotic liver was available. Hepatic tissue obtained in 12 healthy livers was used as control. CD90 gene expression was studied by RT-qPCR, protein expression was assessed by quantitative Western Blot, immunofluorescence and flow cytometry. The CD90 expression analysis showed a significant increment in tumor compared to both its paired cirrhotic tissue and normal liver (p<0.05 and p<0.001, respectively). This increase was accompanied by the up-regulation of stromal component in the cancer, as demonstrated by alpha smooth muscle actin staining. In vitro analysis of JHH-6 cell line showed a higher proliferation capacity of CD90+ compared to CD90- cells (p<0.001), also noticed in 3D clonogenic assay (p<0.05), associated by a significant higher expression of the promoting factors (hepatocyte growth factor, fibroblast associated protein and alpha smooth muscle actin 2). A higher expression of the breast cancer resistance protein was found in CD90+ subpopulation while the multidrug resistance protein 1 showed an opposite behavior. Collectively, these results point to the importance of CD90 in the HCC.
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Funding: CHCS was supported by fellowships of the Italian Ministry of Foreign Affairs of the Istituto Italiano di Cultura, Jakarta, Indonesia, Fondazione
Umberto Veronesi, Milan, Italy, and Fondazione Aldo Duca of the University of Trieste, Italy; BA fellowship was partially supported by grant PORFESR
2007/2013 (2000/ISTR/2011); This work was supported by a grant from the Italian Liver Foundation. The funders had nor role in study design, data
collection and analysis, decision to publish, or preparation of the manuscripts. No additional external funding received for this study.
Competing interests: The authors declare they have no competing interests as defined by PLoS ONE or other interests that might be perceived to
influence the results and discussion reported in this paper.
Primary liver cancer (PLC) is the fifth most common
neoplasms in the world and the third most common cause of
cancer-related death. PLC accounts for around 1% of all death
worldwide [1]. Approximately more than 500,000 new cases
are diagnosed per year, with an age-adjusted worldwide
incidence of 5.514.9 per 100,000 populations [2].
Hepatocellular carcinoma (HCC) accounts for around 85-95%
of all PLC cases. The development of HCC is usually derived
as a final consequence of long term liver injury, in which liver
cirrhosis takes part as the strongest risk factor [3,4]. Most of
HCC patients with intermediate-advanced stages receive
systemic chemotherapies, such as conventional
chemoembolization with doxorubicin [5].
The CD90 (Thy-1) had been proposed as one of the
important molecules in cancer, including in HCC. CD90 is a
25-37 kDa glycophosphatidylinositol (GPI)-anchored protein
expressed in many cells such as T-cells, thymocytes, neurons,
endothelial cells and fibroblast. CD90 operates as an important
regulator of cell to cell and cell to matrix interaction, apoptosis,
adhesion, migration, cancer and fibrosis [6]. CD90 is also
expressed in bone-marrow derived stem cells [7], hepatic stem/
progenitor cells both in adult or fetal livers, but not in adult
hepatocytes [810]. Cells with phenotype
CD90+CD44+CD29+CD73+ isolated from normal adult liver
show osteogenic and endothelial potential differentiation, and
could be also induced to pancreatic islet-like structures [8]. In
prostate cancer, an increased expression of CD90 was
associated with the presence of cancer associated fibroblasts
(CAFs) in the tumor microenvironment and served as cancer
biomarker [11,12].
In liver, CD90 expression was found preferably in poorly
differentiated HCC an (...truncated)