Pharmaceutical Research

, Volume 31, Issue 11, pp 3106–3119 | Cite as

Evaluating the Anticancer Properties of Liposomal Copper in a Nude Xenograft Mouse Model of Human Prostate Cancer: Formulation, In Vitro, In Vivo, Histology and Tissue Distribution Studies

  • Yan Wang
  • San Zeng
  • Tien-Min Lin
  • Lisa Krugner-Higby
  • Doug Lyman
  • Dana Steffen
  • May P. XiongEmail author
Research Paper



Although Cu complexes have been investigated as anticancer agents, there has been no description of Cu itself as a cancer killing agent. A stealth liposomal Cu formulation (LpCu) was studied in vitro and in vivo.


LpCu was evaluated in prostate cancer origin PC-3 cells by a metabolic cytotoxicity assay, by monitoring ROS, and by flow cytometry. LpCu efficacy was evaluated in vivo using intratumoral and intravenous injections into mice bearing PC-3 xenograft tumors. Toxicology was assessed by performing hematological and blood biochemistry assays, and tissue histology and Cu distribution was investigated by elemental analysis.


LpCu and free Cu salts displayed similar levels of cell metabolic toxicity and ROS. Flow cytometry indicated that the mechanisms of cell death were both apoptosis and necrosis. Animals injected i.t. with 3.5 mg/kg or i.v. with 3.5 and 7.0 mg/kg LpCu exhibited significant tumor growth inhibition. Kidney and eye were the main organs affected by Cu-mediated toxicities, but spleen and liver were the major organs of Cu deposition.


LpCu was effective at reducing tumor burden in the xenograft prostate cancer model. There was histological evidence of Cu toxicity in kidneys and eyes of animals treated at the maximum tolerated dose of LpCu 7.0 mg/kg.


liposomes copper reactive oxygen species cancer therapy toxicity 



Alanine aminotransferase


Aspartate aminotransferase


Bicinchoninic acid






Dilution factor


Dynamic light scattering


Dulbecco’s modified eagle medium


Dulbecco’s phosphate buffered saline


Enhanced permeability and retention


Fetal bovine serum


Hank’s balanced salt solution


2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid buffer






Stealth formulation of liposomal Cu


Large unilamellar vesicle


Multilamellar vesicle


Maximum tolerated dose


Phosphate buffered saline


Polydispersity index


Reactive oxygen species


Tert-butyl hydroperoxide


Wisconsin veterinary diagnostic laboratory



This research was supported by NIH grants R00CA136970 and R01DK099596, and startup funds from the University of Wisconsin-Madison, School of Pharmacy. We are grateful to Professor Manish Patankar and Dr. Arvinder Kapur (Dept. of Ob/Gyn, University of Wisconsin-Madison), and the UWCCC Flow Cytometry Laboratory for invaluable assistance with all the flow cytometry assays. We would also like to thank the WVDL Chemistry Section for performing all the elemental tissue analyses.

Supplementary material

11095_2014_1403_MOESM1_ESM.docx (8.5 mb)
ESM 1 (DOCX 8748 kb)


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Yan Wang
    • 1
  • San Zeng
    • 1
  • Tien-Min Lin
    • 1
  • Lisa Krugner-Higby
    • 2
  • Doug Lyman
    • 3
  • Dana Steffen
    • 1
  • May P. Xiong
    • 1
    Email author
  1. 1.School of PharmacyUniversity of Wisconsin – MadisonMadisonUnited State of America
  2. 2.Research Animal Resources CenterUniversity of Wisconsin-MadisonMadisonUnited State of America
  3. 3.Wisconsin Veterinary Diagnostic LaboratoryMadisonUnited State of America

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