Characterisation of the Cell Line HC-AFW1 Derived from a Pediatric Hepatocellular Carcinoma

Abstract

Current treatment of paediatric hepatocellular carcinoma (HCC) is often inefficient due to advanced disease at diagnosis and resistance to common drugs. The aim of this study was to generate a cell line derived from a paediatric HCC in order to expand research in this field. We established the HC-AFW1 cell line from a liver neoplasm of a 4-year-old boy through culturing of primary tumor specimens. The cell line has been stable for over one year of culturing and has a doubling time of 40 h. The tumour cells have an epithelial histology and express HCC-associated proteins such as Alpha-fetoprotein (AFP), Glypican 3, E-cadherin, CD10, CD326, HepPar1 and Vimentin. Forty-nine amino acids in exon 3 of β-Catenin that involve the phosphorylation sites of GSK3 were absent and β-Catenin is detectable in the cell nuclei. Cytogenetic analysis revealed large anomalies in the chromosomal map. Several alterations of gene copy numbers were detected by genome-wide SNP array. Among the different drugs tested, cisplatin and irinotecan showed effective inhibition of tumour cell growth in a proliferation assay at concentrations below 5 µg/ml. Subcutaneous xenotransplantation of HC-AFW1 cells into NOD/SCID mice resulted in fast growing dedifferentiated tumours with high levels of serum AFP. Histological analyses of the primary tumour and xenografts included national and international expert pathological review. Consensus reading characterised the primary tumour and the HC-AFW1-derived tumours as HCC. HC-AFW1 is the first cell line derived from a paediatric HCC without a background of viral hepatitis or cirrhosis and represents a valuable tool for investigating the biology of and therapeutic strategies for childhood HCC.

Activation of β-Catenin and Yap1 in Human Hepatoblastoma and Induction of Hepatocarcinogenesis in Mice

“…In fact HepG2 cells show deletion of 116 and HC-AFW1 a deletion of 49 amino acids. 15,16 Successful silencing of β-catenin or Yap gene in these two cell lines (Figure 2A) led to a notable decrease in cell proliferation and a significant increase in apoptosis as well (Figure 2B-C). However, more pronounced and synergistic anti-growth effect was observed following simultaneous silencing of β-catenin and Yap genes in HepG2 and HC-AFW1 cells (Figure 2B-C).…”

Clinicopathological significance and prognostic value of the expression of the cancer stem cell marker CD133 in hepatocellular carcinoma: a meta-analysis

“…Using CD133 as a biomarker is also limited in terms of predicting clinicopathological significance and prognosis in HCC patients. In addition to CD133, some other cell surface molecules have been considered potential CSC markers in HCC patients, including CD90, CD44, and EpCAM [44,45]. Ma et al found that CD133 + ALDH + cells were significantly more tumorigenic than CD133 + ALDH − or CD133 − ALDH − cells both in vitro and in vivo [46].…”

Anti-tumor activity of sorafenib in a model of a pediatric hepatocellular carcinoma

“…Cell lines and culture conditions HC-AFW1 was derived from a 5-year old boy suffering from HCC [11]. Huh7, a cell line derived from a differentiated aHCC, was obtained from the Japanese Collection of Research Bioresources (JCRB0403).…”

“…Viability was measured using MTT assay as described above. Sensitization was further investigated combining sorafenib with cisplatin (3.9 mg/ml) and doxorubicin (4.0 mg/ml), using IC50 doses as previously described [11].…”

“…Thus a preclinical evaluation may enable a rational selection of new therapeutic concepts. Recently, we described the successful establishment of the continuous epithelial cell line HC-AFW1 derived from a pHCC without viral background [11]. This unique model for pHCC renders a systematic analysis of different treatment strategies in this tumor entity possible.…”