Establishment and Characterization of a New Intrahepatic Cholangiocarcinoma Cell Line Resistant to Gemcitabine

Abstract

Intrahepatic cholangiocarcinoma (ICC) is one of the most lethal liver cancers. Late diagnosis and chemotherapy resistance contribute to the scarce outfit and poor survival. Resistance mechanisms are still poorly understood. Here, we established a Gemcitabine (GEM) resistant model, the MT-CHC01R1.5 cell line, obtained by a GEM gradual exposure (up to 1.5 µM) of the sensitive counterpart, MT-CHC01. GEM resistance was irreversible, even at high doses. The in vitro and in vivo growth was slower than MT-CHC01, and no differences were highlighted in terms of migration and invasion. Drug prediction analysis suggested that Paclitaxel and Doxycycline might overcome GEM resistance. Indeed, in vitro MT-CHC01R1.5 growth was reduced by Paclitaxel and Doxycycline. Importantly, Doxycycline pretreatment at very low doses restored GEM sensitivity. To assess molecular mechanisms underlying the acquisition of GEM resistance, a detailed analysis of the transcriptome in MT-CHC01R1.5 cells versus the corresponding parental counterpart was performed. Transcriptomic analysis showed that most up-regulated genes were involved in cell cycle regulation and in the DNA related process, while most down-regulated genes were involved in the response to stimuli, xenobiotic metabolism, and angiogenesis. Furthermore, additional panels of drug resistance and epithelial to mesenchymal transition genes (n = 168) were tested by qRT-PCR and the expression of 20 genes was affected. Next, based on a comparison between qRT-PCR and microarray data, a list of up-regulated genes in MT-CHC01R1.5 was selected and further confirmed in a primary cell culture obtained from an ICC patient resistant to GEM. In conclusion, we characterized a new GEM resistance ICC model that could be exploited either to study alternative mechanisms of resistance or to explore new therapies.

Omics-Based Platforms: Current Status and Potential Use for Cholangiocarcinoma

“…Peraldo Neia et al revealed the gene and microRNA pattern of the iCCA patient-derived xenograft (PDX) model (GSE84918/GSE84938) [ 85 ]. Varamo analyzed the specific signatures to indicate the gemcitabine resistance CCA cell lines (GSE116118) [ 23 ]. Long-term exposure of gemcitabine-resistance cell lines (>1.5 uM, nine months) compared with sensible counterpart cells.…”

“…In addition, they also found new molecules with differently expressed in their model (QPRT, RRM1, FOSB, PRSS1, etc.) [ 23 ]. Moreover, Gu et al tried to find the linkage between microcystin–leucine–arginine (MC-LR) and iCCA (GSE140687).…”

“…However, the response to such therapy remains limited [ 22 ]. Most importantly, CCA can escape several known tolerance mechanisms induced by chemotherapy (including cisplatin, gemcitabine and other combinations) [ 23 ]. Drug development that relies on well-known signal transductions needs in-depth discussion.…”

Recreating Tumour Complexity in a Dish: Organoid Models to Study Liver Cancer Cells and their Extracellular Environment

“…For example, a wide panel of 25 HCC cell lines was used to investigate the relationships between HCC molecular subtype and specific drug responses [37]. Research with 2D cell lines is still ongoing, with a particular focus on establishing novel cell lines with unique aspects that are not well-represented in previous cell lines (e.g., gemcitabine resistance in iCCA [38] or a highly tumorigenic HCC) [39].…”

Antitumor Efficacy of the Herbal Recipe Benja Amarit against Highly Invasive Cholangiocarcinoma by Inducing Apoptosis both In Vitro and In Vivo

“…In Thailand, CCA is highly prevalent and is the predominant pathological type, accounting for more than 80% of all detected primary liver cancer [ 2 , 3 ]. Due to its aggressiveness and the poor prognosis often given to patients suffering from this disease, CCA is still associated with high mortality rates, especially in the northeast of Thailand [ 1 , 4 , 5 ]. At present, only a quarter of CCA patients are eligible for surgical resection of the tumor, while most of them are prescribed chemotherapy as a form of treatment [ 6 ].…”