Establishment and characterization of two 5-fluorouracil-resistant hepatocellular carcinoma cell lines

Abstract

5-Fluorouracil (5-FU) chemotherapy is the first choice treatment for advanced hepatocellular carcinoma (HCC), and resistance is the major obstacle to successful treatment. Recent studies have reported that epithelial-to-mesenchymal transition (EMT) is associated with chemoresistance in cancers. We speculated that EMT and 5-FU metabolism are related to the mechanism of 5-FU resistance. First, two 5-FU-resistant cell lines, HLF-R4 and HLF-R10, were established from the HLF undifferentiated human HCC cell line. Whereas cell growth was similar in the HLF and HLF-R cell lines, HLF-Rs are about 4- and 10-fold more resistant compared with the HLF cells; thus, we named these cell lines HLF-R4 and HLF-R10, respectively. The terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling assay also showed a dramatically decreased number of apoptotic cells in the HLF-Rs after treatment with 5-FU. We next assessed the characteristics of the HLF, HLF-R4 and HLF-R10 cells. Consistent with our hypothesis, the HLF-Rs had typical morphologic phenotypes of EMT, loss of cell-cell adhesion, spindle-shaped morphology and increased formation of pseudopodia. Real-time quantitative reverse transcriptase polymerase chain reaction data showed downregulated E-cadherin and upregulated Twist-1 and also indicated that EMT changes occurred in the HLF-Rs. We also found decreased ribonucleotide reductase and increased multidrug resistance protein 5 genes in the HLF-R cells. Our results suggested that the metabolism of EMT and 5-FU has important roles in 5-FU chemoresistance in the HLF-R cells, and that the HLF-R cells would be useful in vitro models for understanding the 5-FU-resistant mechanisms in HCC.

Characterization and Resistance Mechanisms of A 5-fluorouracil-resistant Hepatocellular Carcinoma Cell Line

“…The RI of our established HCC cell line is a moderate value of 18 which might be more similar to actual RIs for HCC clinically. Our results were consistent with another two 5-FU-resistant HCC cell lines with 4-and 10-fold RIs (Uchibori et al, 2012). Besides the RI values, the time to development of MDR (10 months), and the maximum concentration of 5-FU to maintain the MDR phenotype (50 µmol/L) have more clinical similarities to clinical practice, which indicate that the BEL-7402/5-FU cells established by us could reflect the MDR development accurately for HCC patients received chemotherapy with 5-FU.…”

Development and characterization of a hydrogen peroxide-resistant cholangiocyte cell line: A novel model of oxidative stress-related cholangiocarcinoma genesis

“…Interestingly, ox-MMNK1-L cells showed the formation of pseudopodia and the loss of cell-to-cell adhesion (multi-layers) under additional oxidative stress. These changes are typical of the epithelial-to-mesenchymal transition [32,33]. Taken together, our results indicate that ox-MMNK1-L cells have acquired several properties of cancer cells.…”

Potential molecular, cellular and microenvironmental mechanism of sorafenib resistance in hepatocellular carcinoma

“…As is well known, EMT is of paramount relevance for various developmental processes, including gastrulation, neural crest formation and carcinoma progression. In the last decades, EMT has been extensively studied and validated in the progression of various carcinomas such as HCC [23,24]. A large body of researches has claimed the important role of EMT in facilitating tumor development and progression by driving metastasis, through the acquisition of enhanced migratory and invasive potential [25][26][27][28][29][30].…”