Establishment and biological characterization of a novel cell line derived from hepatoid adenocarcinoma originated at the ampulla of Vater

Abstract

Hepatoid adenocarcinoma is a rare gastrointestinal tumor and mostly reported in the stomach. Effective chemotherapy has yet to be developed to improve poor prognosis. The present study was undertaken to establish a useful cell line derived from a hepatoid adenocarcinoma, possibly leading to a new therapeutic strategy. The new human cell line VAT-39 was established from a metastatic lymph node of a 69-year-old Japanese male patient with hepatoid adenocarcinoma of the ampulla of Vater. The primary tumor and metastatic lymph node were composed of hepatoid adenocarcinoma cells exhibiting immunohistochemical reactivity for alpha-fetoprotein (AFP) and glypican-3 (GPC3). In the metastatic lymph node, Periodic acid-Schiff (PAS) staining clarified diffuse deposition of glycogen in the cytoplasm, indicating analogous characteristics to the primary hepatoid adenocarcinoma. Moreover, VAT-39 cells produced high levels of AFP in the cultured medium, and reverse-transcriptase polymerase chain reaction (RT-PCR) verified increased expression of GPC3 mRNA in this cell line. Further, we evaluated the sensitivity to major chemotherapeutic drugs against the bile duct cancer. Neither 5-fluorouracil nor gemcitabine showed particular sensitivity to this cell line. The tumorigenicity of the cultured cells was confirmed in athymic nude mice and the histological features of the explanted tumor were similar to the VAT-39 cell line. The present VAT-39 is the first hepatoid adenocarcinoma cell line that originates from the ampulla of Vater and it will be applicable for basic biological studies searching for new strategies of molecular targeted chemotherapy to this disease.

The HDAC Inhibitor, SAHA, Combined with Cisplatin Synergistically Induces Apoptosis in Alpha-fetoprotein-producing Hepatoid Adenocarcinoma Cells

“…VAT-39 human HAC cells from the ampulla of Vater [33] were maintained in Dulbecco’s Modified Eagle’s Medium with L-glutamine and phenol red (Fujifilm Wako Pure Chemical Corporation, Osaka, Japan). The medium was supplemented with 10% (v/v) heat-inactivated fetal bovine serum and penicillin-streptomycin-amphotericin B in a humidified atmosphere containing 5% CO 2 at 37°C.…”

“…Culture medium was harvested at 48 hr, and the AFP concentration was measured with an Architect AFP assay using an Architecto i200SR (Abbott, Chiba, Japan) [33].…”

“…Most cases occur in the stomach, but HAC is also found in the lung, pancreas, esophagus, ampulla of Vater, colon, urinary bladder, renal pelvis, ovary, and cervix [31, 34]. Clinically, HAC shows aggressive progression and a strong tendency to metastasize to lymph nodes and liver; and AFP production promotes neovascularization, high proliferative activity, and inhibition of apoptosis, which contribute to the poor prognosis [20, 33]. A recent meta-analysis found a 3-year survival rate of 7.36% and a median survival time of 10 months, indicating a poor prognosis [25].…”

“…In this study, we investigated the effects of a combination of cisplatin and SAHA in VAT-39 cells, a newly established HAC cell line [33]. Cell viability and apoptosis were examined by MTT assay, flow cytometry, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay.…”

Establishment and characterization of an immortalized human hepatocyte line for the development of bioartificial liver system

“…Cells grown on coverslips were fixed with 3% formaldehyde for 20 min, then the coverslips were stained using a PAS staining kit (Maixin. Bio, Fuzhou, China) following the manufacturer's instructions (Saraiva-Pava et al 2015;Takahashi et al 2014). The PAS-stained cells were counterstained with haematoxylin for 6 min, and finally examined under a light microscopy (IX73, Olympus, Beijing, China).…”