Knockdown of the cytoprotective chaperone, clusterin, chemosensitizes human breast cancer cells both in vitro and in vivo

Mol Cancer Ther. 2005 Dec;4(12):1837-49. doi: 10.1158/1535-7163.MCT-05-0178.

Abstract

Clusterin is a stress-associated cytoprotective chaperone up-regulated by various apoptotic triggers in many cancers and confers treatment resistance when overexpressed. The objectives of this study were to evaluate clusterin expression levels in human breast cancer and to determine whether antisense oligonucleotides or double-stranded small interfering RNAs (siRNA) targeting the clusterin gene enhance apoptosis induced by paclitaxel. Clusterin immunostaining was evaluated in a tissue microarray of 379 spotted breast cancers. The effect of hormone withdrawal, paclitaxel treatment, clusterin antisense oligonucleotide (OGX-011), and siRNA treatments on clusterin expression was examined in MCF-7 and MDA-MB-231 cells. Northern, quantitative real-time PCR, and Western analyses were used to measure change in clusterin mRNA and protein levels. The effect of OGX-011 or siRNA clusterin treatment on chemosensitivity to paclitaxel was done in both cell lines in vitro, whereas the ability of OGX-011 to chemosensitize in vivo was evaluated in athymic mice bearing MCF-7 tumors. Clusterin was expressed in 62.5% of tumors within the tissue microarray. Clusterin expression increased after estrogen withdrawal and paclitaxel treatment in vitro in MCF-7 cells. OGX-011 or siRNA clusterin decreased clusterin levels by >90% in a dose-dependent, sequence-specific manner and significantly enhanced chemosensitivity to paclitaxel in vitro. When combined, OGX-011 or siRNA clusterin reduced the IC50 by 2-log compared with paclitaxel alone. In vivo administration of OGX-011 enhanced the effects of paclitaxel to significantly delay MCF-7 tumor growth. These data identify clusterin as a valid therapeutic target and provides preclinical proof-of-principle to test OGX-011 in multimodality therapies for breast cancer.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Blotting, Western
  • Breast Neoplasms / pathology*
  • Cell Division / drug effects
  • Cell Line, Tumor
  • Clusterin / genetics
  • Clusterin / metabolism
  • Clusterin / physiology*
  • Female
  • Humans
  • Middle Aged
  • Oligonucleotides, Antisense / pharmacology*
  • Paclitaxel / pharmacology*
  • Polymerase Chain Reaction
  • RNA, Small Interfering / genetics
  • Thionucleotides / pharmacology*
  • Transplantation, Heterologous

Substances

  • Antineoplastic Agents, Phytogenic
  • Clusterin
  • OGX-011
  • Oligonucleotides, Antisense
  • RNA, Small Interfering
  • Thionucleotides
  • Paclitaxel