In vitro targeting of a cytotoxic analog of luteinizing hormone-releasing hormone AN-207 to ES-2 human ovarian cancer cells as demonstrated by microsatellite analyses

J. M. Arencibia, Andrew V Schally, G. Halmos, A. Nagy, H. Kiaris

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Targeting of cytotoxic agents represents a modern approach to the treatment of various cancers, that improves the efficacy and reduces peripheral toxicity. Recently we developed e powerful cytotoxic analog of luteinizing hormone-releasing hormone (LHRH), AN-207, designed to be targeted to tumors that express LHRH receptors. This analog consists of the superactive derivative of doxorubicin (DOX), 2-pyrrolino-DOX (AN-201), linked to [D-Lys6]LHRH carrier. In the present study we investigated the cytocidal effects of AN-207 and AN-201 on the LHRH receptor-positive ES-2 ovarian cancer cells. The targeting of AN-207 to ES-2 cells in the presence of LHRH receptor-negative UCI-107 ovarian cancer cells was also evaluated by semi-quantitative polymerase chain reaction (PCR) amplification of microsatellite markers. Ligand competition assays showed a single class of high-affinity and low-capacity binding sites in ES-2 cells with a mean dissociation constant (KD) of 3.93±0.1 nM and a mean maximal binding capacity (Bmax) of 271±26.1 fmol/mg membrane protein. Kinetic assays indicated that AN-207 caused cell death in a concentration- and time-dependent manner in ES-2 cells, but not in UCI-107 cells, while the kinetics of cytotoxic effects of AN-201 were similar in both cell lines. To investigate targeting, ES-2 cells were co-cultured with UCI-107 cells, treated with 10 nM AN-207 or AN-201 for different times and then cultured for 48 h in the absence of cytotoxic agents. Genomic DNA was extracted for microsatellite analyses using different markers. Semi-quantitative analyses of the intensity of the alleles that correspond to each cell line indicated that AN-207 was selectively targeted to ES-2 cells, while AN-201 showed no selectivity for either cell line. These results extend our previous findings that AN-207 can be targeted to ovarian cancers and other tumors that express receptors for LHRH. Cytotoxic analogs of LHRH, such as AN-207, should be considered for treatment of LHRH receptor-positive tumors.

Original languageEnglish
Pages (from-to)71-78
Number of pages8
JournalAnti-Cancer Drugs
Volume12
Issue number1
DOIs
StatePublished - Feb 20 2001
Externally publishedYes

Fingerprint

Gonadotropin-Releasing Hormone
Microsatellite Repeats
Ovarian Neoplasms
LHRH Receptors
Cytotoxins
Cell Line
Doxorubicin
Neoplasms
AN 207
In Vitro Techniques
Membrane Proteins
Cell Death
Alleles
Binding Sites
AN 204
Ligands
Polymerase Chain Reaction
DNA

Keywords

  • Cytotoxic LHRH analog
  • LHRH receptor
  • Ovarian cancer
  • Targeted cancer therapy

ASJC Scopus subject areas

  • Pharmacology
  • Cancer Research
  • Oncology

Cite this

In vitro targeting of a cytotoxic analog of luteinizing hormone-releasing hormone AN-207 to ES-2 human ovarian cancer cells as demonstrated by microsatellite analyses. / Arencibia, J. M.; Schally, Andrew V; Halmos, G.; Nagy, A.; Kiaris, H.

In: Anti-Cancer Drugs, Vol. 12, No. 1, 20.02.2001, p. 71-78.

Research output: Contribution to journalArticle

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