V-ATPase inhibition by archazolid leads to lysosomal dysfunction resulting in impaired cathepsin B activation in vivo

Int J Cancer. 2014 May 15;134(10):2478-88. doi: 10.1002/ijc.28562. Epub 2013 Nov 14.

Abstract

The myxobacterial agent archazolid inhibits the vacuolar proton pump V-ATPase. V-ATPases are ubiquitously expressed ATP-dependent proton pumps, which are known to regulate the pH in endomembrane systems and thus play a crucial role in endo- and exocytotic processes of the cell. As cancer cells depend on a highly active secretion of proteolytic proteins in order to invade tissue and form metastases, inhibition of V-ATPase is proposed to affect the secretion profile of cancer cells and thus potentially abrogate their metastatic properties. Archazolid is a novel V-ATPase inhibitor. Here, we show that the secretion pattern of archazolid treated cancer cells includes various prometastatic lysosomal proteins like cathepsin A, B, C, D and Z. In particular, archazolid induced the secretion of the proforms of cathepsin B and D. Archazolid treatment abrogates the cathepsin B maturation process leading to reduced intracellular mature cathepsin B protein abundance and finally decreased cathepsin B activity, by inhibiting mannose-6-phoshate receptor-dependent trafficking. Importantly, in vivo reduced cathepsin B protein as well as a decreased proteolytic cathepsin B activity was detected in tumor tissue of archazolid-treated mice. Our results show that inhibition of V-ATPase by archazolid reduces the activity of prometastatic proteases like cathepsin B in vitro and in vivo.

Keywords: V-ATPase; archazolid; cancer; cathepsin B; metastasis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Cathepsin B / genetics
  • Cathepsin B / metabolism*
  • Cell Line, Tumor
  • Enzyme Activation / drug effects
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Lysosomes / drug effects
  • Lysosomes / enzymology
  • MCF-7 Cells
  • Macrolides / pharmacology*
  • Mice
  • Mice, Inbred BALB C
  • Neoplasms / genetics
  • Neoplasms / metabolism
  • Neoplasms / pathology
  • Neoplasms, Experimental / genetics
  • Neoplasms, Experimental / metabolism
  • Neoplasms, Experimental / prevention & control
  • Protein Transport / drug effects
  • RNA Interference
  • Receptor, IGF Type 2 / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Thiazoles / pharmacology*
  • Tumor Burden / drug effects
  • Vacuolar Proton-Translocating ATPases / antagonists & inhibitors*
  • Vacuolar Proton-Translocating ATPases / genetics
  • Vacuolar Proton-Translocating ATPases / metabolism

Substances

  • ATP6V0C protein, human
  • Macrolides
  • Receptor, IGF Type 2
  • Thiazoles
  • archazolid B
  • cation-dependent mannose-6-phosphate receptor
  • Cathepsin B
  • Vacuolar Proton-Translocating ATPases