Establishment of an Allo-Transplantable Hamster Cholangiocarcinoma Cell Line and Its Application for In Vivo Screening of Anti-Cancer Drugs

Abstract

Opisthorchis viverrini (O. viverrini) is a well-known causative agent of cholangiocarcinoma (CCA) in humans. CCA is very resistant to chemotherapy and is frequently fatal. To understand the pathogenesis of CCA in humans, a rodent model was developed. However, the development of CCA in rodents is time-consuming and the xenograft-transplantation model of human CCA in immunodeficient mice is costly. Therefore, the establishment of an in vivo screening model for O. viverrini-associated CCA treatment was of interest. We developed a hamster CCA cell line, Ham-1, derived from the CCA tissue of O. viverrini-infected and N-nitrosodimethylamine-treated Syrian golden hamsters. Ham-1 has been maintained in Dulbecco's Modified Essential Medium supplemented with 10% fetal bovine serum for more than 30 subcultures. These cells are mostly diploid (2n=44) with some being polyploid. Tumorigenic properties of Ham-1 were demonstrated by allograft transplantation in hamsters. The transplanted tissues were highly proliferative and exhibited a glandular-like structure retaining a bile duct marker, cytokeratin 19. The usefulness of this for in vivo model was demonstrated by berberine treatment, a traditional medicine that is active against various cancers. Growth inhibitory effects of berberine, mainly by an induction of G1 cell cycle arrest, were observed in vitro and in vivo. In summary, we developed the allo-transplantable hamster CCA cell line, which can be used for chemotherapeutic drug testing in vitro and in vivo.

Anticancer effect of berberine based on experimental animal models of various cancers: a systematic review and meta-analysis

“…Study characteristics are summarized in Table 1. A total of 26 studies [10–35] from 2000 to 2018, focusing on various cancer types, including breast cancer [10–16], liver cancer [17–19], colorectal cancer [20–22], nasopharyngeal carcinoma [23, 24], lung cancer [25, 26], gastric cancer [27, 28], neuroepithelial tumor [29, 30], endometrial carcinoma [31], esophageal cancer [32], tongue cancer [33], cholangiocarcinoma [34], and sarcoma [35] were included. The studies used rats [14], hamsters [34], and mice [10–13, 15–33, 35] modeled via subcutaneous tumor implantation [10–13, 16–35] or induced tumor formation [14, 15].…”

“…A total of 26 studies [10–35] from 2000 to 2018, focusing on various cancer types, including breast cancer [10–16], liver cancer [17–19], colorectal cancer [20–22], nasopharyngeal carcinoma [23, 24], lung cancer [25, 26], gastric cancer [27, 28], neuroepithelial tumor [29, 30], endometrial carcinoma [31], esophageal cancer [32], tongue cancer [33], cholangiocarcinoma [34], and sarcoma [35] were included. The studies used rats [14], hamsters [34], and mice [10–13, 15–33, 35] modeled via subcutaneous tumor implantation [10–13, 16–35] or induced tumor formation [14, 15]. BBR was administered in doses ranging between 2.5 mg/kg and 200 mg/kg body weight through intraperitoneal injection [10, 15–17, 19, 21, 23, 24, 30, 33, 35] and gavage [11–14, 18, 20, 22, 25–29, 31, 32, 34] or from 1000 ppm to 5400 ppm in drinking water [13, 20, 25].…”

“…The studies used rats [14], hamsters [34], and mice [10–13, 15–33, 35] modeled via subcutaneous tumor implantation [10–13, 16–35] or induced tumor formation [14, 15]. BBR was administered in doses ranging between 2.5 mg/kg and 200 mg/kg body weight through intraperitoneal injection [10, 15–17, 19, 21, 23, 24, 30, 33, 35] and gavage [11–14, 18, 20, 22, 25–29, 31, 32, 34] or from 1000 ppm to 5400 ppm in drinking water [13, 20, 25]. The size of the study sample ranged from 6 to 20, while the follow-up ranged from 1 week to 32.5 weeks.…”

Establishment of Highly Transplantable Cholangiocarcinoma Cell Lines from a Patient-Derived Xenograft Mouse Model

“…For cell line development, fresh xenograft tissues were prepared as previously described [34]. Cells were cultured in DMEM/F12 (Wako) containing 1–10% FBS and insulin-transferrin-selenium (ITS, Gibco BRL, Carlsbad, CA, USA).…”

Preventive and Therapeutic Roles of Berberine in Gastrointestinal Cancers

“…Many studies have demonstrated that the therapeutic effect of BBR on cholangiocarcinoma and HCC occurs via inhibiting cancer cell proliferation and promoting cancer cell apoptosis. Puthdee et al demonstrated that the inhibitory effects of BBR on the proliferation of hamster cholangiocarcinoma cells in vitro and in vivo occurred via the induction of cell cycle arrest in the G1 phase[41].Li et al reported that BBR induces the cell cycle arrest of HCC cells in the G0/G1 phase by enhancing CDKIs p21Cip1 and p27Kip1 expression via Akt/FoxO3a/Skp2 axis regulation[42]. Saxena et al showed that BBR induces mitochondrial impairment and apoptosis of human hepatoma cells by modulating the PHLPP2-Akt-MST1 kinase signaling pathway[43].…”