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Enhanced gemcitabine cytotoxicity with knockdown of multidrug resistance protein genes in human cholangiocarcinoma cell lines
Author(s) -
Yang Jiaqi,
Sontag David,
Gong Yuewen,
Minuk Gerald Y
Publication year - 2021
Publication title -
journal of gastroenterology and hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.214
H-Index - 130
eISSN - 1440-1746
pISSN - 0815-9319
DOI - 10.1111/jgh.15289
Subject(s) - gene knockdown , gemcitabine , cytotoxicity , multidrug resistance associated protein 2 , small hairpin rna , cancer research , biology , multiple drug resistance , downregulation and upregulation , cell culture , medicine , drug resistance , chemotherapy , gene , microbiology and biotechnology , atp binding cassette transporter , in vitro , biochemistry , genetics , transporter
Background and Aim Cholangiocarcinoma (CCA) is an often fatal primary cancer of the liver that tends to be resistant to chemotherapy. Multidrug resistance proteins (MRPs) contribute to the chemoresistance of these tumors. The objectives of the study were to document MRP expression profiles in two representative human intrahepatic and extrahepatic CCA cells lines (HuCCT1 and KMBC, respectively) and gemcitabine‐induced cytotoxicity prior to and following MRP knockdown. Methods Multidrug resistance protein mRNA and protein expression were documented by real‐time reverse transcription–polymerase chain reaction and western blots, respectively. MRP knockdown was achieved with lentivirus small hairpin RNA constructs. Results Prior to gemcitabine exposure, MRP1, MRP2, MRP4, MRP5, and MRP6 mRNA were expressed in HuCCT1 cells and MRP1, MRP3, MRP4, and MRP5 in KMBC cells. Following gemcitabine exposure, MRP5 and MRP6 expressions were significantly upregulated in HuCCT1 cells and MRP5 in KMBC cells. In HuCCT1 cells, although MRP5 knockdown had no effect, MRP6 knockdown significantly increased gemcitabine‐induced cytotoxicity. In KMBC cells, MRP5 knockdown significantly increased gemcitabine cytotoxicity. Conclusions Inhibition of MRP6 expression in intra‐hepatic and MRP5 in extra‐hepatic should be explored as potential treatments for CCA in humans.