Establishment and characterization of a new intrahepatic cholangiocarcinoma cell line resistant to gemcitabine

Chiara Varamo, Caterina Peraldo-Neia, Paola Ostano, Marco Basiricò, Chiara Raggi, Paola Bernabei, Tiziana Venesio, Enrico Berrino, Massimo Aglietta, Francesco Leone, Giuliana Cavalloni

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Article number519
JournalCancers
Volume11
Issue number4
DOIs
Publication statusPublished - Apr 1 2019

Fingerprint

gemcitabine
Cholangiocarcinoma
Cell Line
Doxycycline
Paclitaxel
Genes
Polymerase Chain Reaction
Epithelial-Mesenchymal Transition
Primary Cell Culture
Delayed Diagnosis
Gene Expression Profiling
Xenobiotics
Liver Neoplasms
Growth
Drug Resistance
Cell Cycle

Keywords

  • Cholangiocarcinoma
  • Drug resistance
  • Gemcitabine
  • Gene expression profiling
  • Preclinical models

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Establishment and characterization of a new intrahepatic cholangiocarcinoma cell line resistant to gemcitabine. / Varamo, Chiara; Peraldo-Neia, Caterina; Ostano, Paola; Basiricò, Marco; Raggi, Chiara; Bernabei, Paola; Venesio, Tiziana; Berrino, Enrico; Aglietta, Massimo; Leone, Francesco; Cavalloni, Giuliana.

In: Cancers, Vol. 11, No. 4, 519, 01.04.2019.

Research output: Contribution to journalArticle

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AU - Peraldo-Neia, Caterina

AU - Ostano, Paola

AU - Basiricò, Marco

AU - Raggi, Chiara

AU - Bernabei, Paola

AU - Venesio, Tiziana

AU - Berrino, Enrico

AU - Aglietta, Massimo

AU - Leone, Francesco

AU - Cavalloni, Giuliana

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AB - 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.

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