Antisense oligodeoxynucleotides for urokinase-plasminogen activator receptor have anti-invasive and anti-proliferative effects in vitro and inhibit spontaneous metastases of human melanoma in mice

Silvia D'Alessio, Francesca Margheri, Marco Pucci, Angela Del Rosso, Brett P. Monia, Mauro Bologna, Carlo Leonetti, Marco Scarsella, Gabriella Zupi, Gabriella Fibbi, Mario Del Rosso

Research output: Contribution to journalArticle

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Abstract

We have targeted the urokinase-type plasminogen activator receptor (uPAR) with phosphorothioate antisense oligonucleotides (aODN) in vitro to evaluate the anti-invasive and anti-proliferative effects of uPAR down-regulation, as well as in vivo to evaluate anti-tumor and anti-metastatic activity. aODN-dependent uPAR downregulation in vitro was induced in cells of human melanoma, mammary carcinoma, ovarian carcinoma and SV-40-transformed embryonic lung fibroblasts. uPAR was determined by an antibody-based assay and by semiquantitative polymerase chain reaction. Cell invasion was evaluated by Matrigel invasion assay and cell proliferation by direct cell counting. aODN reduced uPAR, invasion and proliferation in all the treated cell lines. Following aODN treatment, human melanoma cells exhibited a strong decrease of uPAR-dependent ERK1/2 activation and were used in vivo to control metastasis in CD-1 male nude (nu/nu) mice by uPAR aODN injection. 60 mice were injected in the hind leg muscles with a suspension of 106 melanoma cells. After 4 days, when a tumor mass of about 350 mg was evident in all the mice injected, 20 mice were treated i.v. with aODN and 20 with dODN at 0.5 mg/day for 5 consecutive days. Twenty control mice were not treated. A second and third cycle of treatment was administered at 2-day intervals. Treatment with aODN resulted into a 78% reduction of lung metastases and 45% reduction of the primary tumor mass with no loss of body weight. Our results suggest to evaluate the utility of uPAR aODN in controlling the metastatic spreading of human melanoma.

Original languageEnglish
Pages (from-to)125-133
Number of pages9
JournalInternational Journal of Cancer
Volume110
Issue number1
DOIs
Publication statusPublished - May 20 2004

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Urokinase Plasminogen Activator Receptors
Oligodeoxyribonucleotides
Melanoma
Neoplasm Metastasis
Down-Regulation
Phosphorothioate Oligonucleotides
Neoplasms
Lung
In Vitro Techniques
Antisense Oligonucleotides
Leg
Suspensions
Therapeutics
Fibroblasts
Body Weight
Cell Proliferation
Breast Neoplasms
Carcinoma
Cell Line
Muscles

Keywords

  • Antisense oligonucleotides
  • ERK
  • Melanoma
  • Metastasis
  • Urokinase receptor (CD87)

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Antisense oligodeoxynucleotides for urokinase-plasminogen activator receptor have anti-invasive and anti-proliferative effects in vitro and inhibit spontaneous metastases of human melanoma in mice. / D'Alessio, Silvia; Margheri, Francesca; Pucci, Marco; Del Rosso, Angela; Monia, Brett P.; Bologna, Mauro; Leonetti, Carlo; Scarsella, Marco; Zupi, Gabriella; Fibbi, Gabriella; Del Rosso, Mario.

In: International Journal of Cancer, Vol. 110, No. 1, 20.05.2004, p. 125-133.

Research output: Contribution to journalArticle

D'Alessio, Silvia ; Margheri, Francesca ; Pucci, Marco ; Del Rosso, Angela ; Monia, Brett P. ; Bologna, Mauro ; Leonetti, Carlo ; Scarsella, Marco ; Zupi, Gabriella ; Fibbi, Gabriella ; Del Rosso, Mario. / Antisense oligodeoxynucleotides for urokinase-plasminogen activator receptor have anti-invasive and anti-proliferative effects in vitro and inhibit spontaneous metastases of human melanoma in mice. In: International Journal of Cancer. 2004 ; Vol. 110, No. 1. pp. 125-133.
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abstract = "We have targeted the urokinase-type plasminogen activator receptor (uPAR) with phosphorothioate antisense oligonucleotides (aODN) in vitro to evaluate the anti-invasive and anti-proliferative effects of uPAR down-regulation, as well as in vivo to evaluate anti-tumor and anti-metastatic activity. aODN-dependent uPAR downregulation in vitro was induced in cells of human melanoma, mammary carcinoma, ovarian carcinoma and SV-40-transformed embryonic lung fibroblasts. uPAR was determined by an antibody-based assay and by semiquantitative polymerase chain reaction. Cell invasion was evaluated by Matrigel invasion assay and cell proliferation by direct cell counting. aODN reduced uPAR, invasion and proliferation in all the treated cell lines. Following aODN treatment, human melanoma cells exhibited a strong decrease of uPAR-dependent ERK1/2 activation and were used in vivo to control metastasis in CD-1 male nude (nu/nu) mice by uPAR aODN injection. 60 mice were injected in the hind leg muscles with a suspension of 106 melanoma cells. After 4 days, when a tumor mass of about 350 mg was evident in all the mice injected, 20 mice were treated i.v. with aODN and 20 with dODN at 0.5 mg/day for 5 consecutive days. Twenty control mice were not treated. A second and third cycle of treatment was administered at 2-day intervals. Treatment with aODN resulted into a 78{\%} reduction of lung metastases and 45{\%} reduction of the primary tumor mass with no loss of body weight. Our results suggest to evaluate the utility of uPAR aODN in controlling the metastatic spreading of human melanoma.",
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AU - Monia, Brett P.

AU - Bologna, Mauro

AU - Leonetti, Carlo

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AU - Zupi, Gabriella

AU - Fibbi, Gabriella

AU - Del Rosso, Mario

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