Suppression of autophagy by CUB domain-containing protein 1 signaling is essential for anchorage-independent survival of lung cancer cells

Takamasa Uekita, Satoko Fujii, Yuri Miyazawa, Akinori Hashiguchi, Hitosi Abe, Michiie Sakamoto, Ryuichi Sakai

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

CUB (C1r/C1s, urchin embryonic growth factor, BMP1) domain-containing protein 1 (CDCP1) has been implicated in promoting metastasis of cancer cells through several mechanisms, including the inhibition of anoikis, which is cell death triggered by the loss of extracellular matrix interactions. However, the mechanism inhibiting cell death regulated by CDCP1 remains elusive. Inhibition of CDCP1 expression using small interfering RNA (siRNA) induced the cell death of suspended cancer cells without cleaving caspase-3, a marker of apoptosis; cell death was not inhibited by a general caspase inhibitor, suggesting that the loss of CDCP1 induces caspase-independent cell death. In contrast, knockdown of CDCP1 as well as protein kinase Cδ (PKCδ), a downstream effector of CDCP1, in a suspension culture of lung cancer cells resulted in marked induction of membranous microtubule-associated protein 1 light chain 3 (LC3)-II protein, a hallmark of autophagy, and caused the formation of an autophagosome structure visualized using green fluorescent protein-tagged LC3-II. Expression and phosphorylation of exogenous CDCP1 by Fyn kinase reduced the formation of autophagosomes and inhibited phosphorylation of CDCP1 by PP2, a Src kinase inhibitor or inhibited PKCδ by rottlerin, stimulating autophagosome formation. Moreover, death of suspended lung cancer cells induced by CDCP1 siRNA or by PKCδ siRNA was reduced by the autophagy inhibitor 3-methyladenine. These results indicate that CDCP1-PKCδ signaling plays a critical role in inhibiting autophagy, which is responsible for anoikis resistance of lung cancer cells.

Original languageEnglish
Pages (from-to)865-870
Number of pages6
JournalCancer Science
Volume104
Issue number7
DOIs
Publication statusPublished - 2013 Jul

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Autophagy
Lung Neoplasms
Cell Death
Protein Kinase C
Anoikis
Small Interfering RNA
Phosphorylation
Light
Caspase Inhibitors
Microtubule-Associated Proteins
src-Family Kinases
Caspases
Green Fluorescent Proteins
Caspase 3
Extracellular Matrix
Neoplasms
Intercellular Signaling Peptides and Proteins
Suspensions
Phosphotransferases
Protein Domains

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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Suppression of autophagy by CUB domain-containing protein 1 signaling is essential for anchorage-independent survival of lung cancer cells. / Uekita, Takamasa; Fujii, Satoko; Miyazawa, Yuri; Hashiguchi, Akinori; Abe, Hitosi; Sakamoto, Michiie; Sakai, Ryuichi.

In: Cancer Science, Vol. 104, No. 7, 07.2013, p. 865-870.

Research output: Contribution to journalArticle

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