Silencing the expression and function of breast cancer resistance protein in MCF7/MX100 cells by shRNA expressing lentivirus

Caihong Yu, Siyun Xu, Aiming Yu, Yanqing Liu, Haihong Hu, Liping Li, Lushan Yu, Hui Zhou, Huidi Jiang, Su Zeng

Research output: Contribution to journalArticle

Abstract

Overexpression of breast cancer resistance protein (ABCG2/BCRP) in cancer cells may cause tumor resistance to chemotherapeutic drugs. RNA interference (RNAi) can selectively silence the expression of a target gene of interest. In the present study, we aimed to modulate the BCRP expression and examine the functional consequence using RNAi approach. Three siRNAs (siBCRP1, siBCRP2 and siBCRP3) targeting BCRP were evaluated in drugresistant MCF7/MX100 cells overexpressing BCRP. The BCRP expression at the mRNA and protein levels was inhibited by siBCRP2 and siBCRP3 over 90% and 70%, respectively. As a result, the intracellular mitoxantrone accumulation was sharply increased in MCF7/MX100 cells after the transfection. Furthermore, shRNA sequences bearing siBCRP2 and siBCRP3 were cloned into lentiviral expression plasmid (pTRIPZ) to package lentivirus, and MCF7/MX100 cells stably expressing siRNA targeted to human ABCG2/BCRP were established by lentivectormediated gene transfer system. The stable cells exhibited an increased miotxantrone accumulation, among which the BCRP expression at the mRNA level was reduced by LentiBCRP2 and LentiBCRP3 around 72% and 56%, respectively. Moreover, the BCRP expression at the protein level was reduced by 70% and 53%, respectively. Furthermore, the cell lines were used to screen active ingredients in traditional herbal medicines in order to evaluate BCRP substrates or inhibitors. Our data suggested that the BCRP knockdown cell lines could serve as good cell models for preclinical studies.

Original languageEnglish (US)
Pages (from-to)743-750
Number of pages8
JournalJournal of Chinese Pharmaceutical Sciences
Volume23
Issue number11
DOIs
StatePublished - 2014

Keywords

  • BCRP
  • Drug resistance reversal
  • Lentiviral expression system
  • Mitoxantrone
  • RNA interference
  • TCM

ASJC Scopus subject areas

  • Pharmaceutical Science

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