Clathrin-dependent endocytosis of claudin-2 by DFYSP peptide causes lysosomal damage in lung adenocarcinoma A549 cells

Akira Ikari*, Saeko Taga, Ryo Watanabe, Tomonari Sato, Shun Shimobaba, Hiroyuki Sonoki, Satoshi Endo, Toshiyuki Matsunaga, Hideki Sakai, Masahiko Yamaguchi, Yasuhiro Yamazaki, Junko Sugatani

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Abstract Claudins are tight junctional proteins and comprise a family of over 20 members. Abnormal expression of claudins is reported to be involved in tumor progression. Claudin-2 is highly expressed in lung adenocarcinoma tissues and increases cell proliferation, whereas it is not expressed in normal tissues. Claudin-2-targeting molecules such as peptides and small molecules may be novel anti-cancer drugs. The short peptide with the sequence DFYSP, which mimics the second extracellular loop of claudin-2, decreased claudin-2 content in the cytoplasmic fraction of A549 cells. In contrast, it did not affect the content in the nuclear fraction. The decrease in claudin-2 content was inhibited by chloroquine (CQ), a lysosomal inhibitor, but not by MG-132, a proteasome inhibitor. In the presence of DFYSP peptide and CQ, claudin-2 was co-localized with LAMP-1, a lysosomal marker. The DFYSP peptide-induced decrease in claudin-2 content was inhibited by monodancylcadaverine (MDC), an inhibitor of clathrin-dependent endocytosis. DFYSP peptide increased lysosome content and cathepsin B release, and induced cellular injury, which were inhibited by MDC. Cellular injury induced by DFYSP peptide was inhibited by necrostatin-1, an inhibitor of necrotic cell death, but not by Z-VAD-FMK, an inhibitor of apoptotic cell death. Our data indicate that DFYSP peptide increases the accumulation of the peptide and claudin-2 into the lysosome, resulting in lysosomal damage. Claudin-2 may be a new target for lung cancer therapy.

Original languageEnglish
Article number81947
Pages (from-to)2326-2336
Number of pages11
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1848
Issue number10
DOIs
StatePublished - 2015/07/20

Keywords

  • Adenocarcinoma
  • Claudin-2
  • Lung
  • Lysosomal damage

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

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