Signaling pathway activated during apoptosis of the prostate cancer celt line LNCaP: Overexpression of caspase-7 as a new gene therapy strategy for prostate cancer

Marco Marcelli, Glenn R. Cunningham, Margaret Walkup, Zening He, Lydia Sturgis, Carolina Kagan, Roberta Mannucci, Ildo Nicoletti, BaBie Teng, Larry Denner

Research output: Contribution to journalArticle

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Abstract

We studied the molecular mechanisms of apoptosis in the prostate cancer cell line LNCaP and whether overexpression of caspase activity could force this cell line to undergo apoptosis. The inhibitor of phosphomevalonate decarboxylase, sodium phenylacetate, and the protein kinase inhibitor staurosporine induced (a) release of cytochrome c from the mitochondria to the cytosol; (b) reduction in mitochondrial transmembrane potential; (c) proteolytic processing of caspase-3 and -7 but not -2; (d) cleavage of the DEVD substrate and the death substrates poly(ADP-ribose) polymerase and DNA fragmentation factor; and (e) apoptosis. The panspecific inhibitor of caspase activation N-benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (z-VAD- FMK) prevented all of these events except release of mitochondrial cytochrome c into the cytosol. None of these apoptotic signaling events were elicited by staurosporine or sodium phenylacetate treatment of LNCaP-Bcl-2 cells that overexpress the oncoprotein Bcl-2. Because caspase-7 is activated in every model of apoptosis that we have characterized thus far, we wished to learn whether overexpression of this protease could directly cause apoptosis of LNCaP cells. By using a replication-defective adenovirus, overexpression of caspase-7 protein in both LNCaP and LNCaP-Bcl-2 cells was accompanied by induction of cleavage of the DEVD substrate and TUNEL. These studies have demonstrated that caspase-7 and -3 are critical mediators of apoptosis in LNCaP cells. Caspase-7 was proteolytically activated in every model of apoptosis that we have developed, and the overexpression of it induced apoptosis of LNCaP and LNCaP-Bcl-2 cells. Thus, adenoviral-mediated transfer of caspase-7 may offer a new effective approach for the treatment of prostate cancer.

Original languageEnglish (US)
Pages (from-to)382-390
Number of pages9
JournalCancer Research
Volume59
Issue number2
StatePublished - Jan 16 1999
Externally publishedYes

Fingerprint

Caspase 7
Genetic Therapy
Prostatic Neoplasms
Apoptosis
Staurosporine
Cytochromes c
Caspase 3
Cytosol
Cell Line
Caspase Inhibitors
Carboxy-Lyases
Poly(ADP-ribose) Polymerases
Oncogene Proteins
In Situ Nick-End Labeling
DNA Fragmentation
Protein Kinase Inhibitors
Caspases
Adenoviridae
Membrane Potentials
Mitochondria

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Signaling pathway activated during apoptosis of the prostate cancer celt line LNCaP : Overexpression of caspase-7 as a new gene therapy strategy for prostate cancer. / Marcelli, Marco; Cunningham, Glenn R.; Walkup, Margaret; He, Zening; Sturgis, Lydia; Kagan, Carolina; Mannucci, Roberta; Nicoletti, Ildo; Teng, BaBie; Denner, Larry.

In: Cancer Research, Vol. 59, No. 2, 16.01.1999, p. 382-390.

Research output: Contribution to journalArticle

Marcelli, M, Cunningham, GR, Walkup, M, He, Z, Sturgis, L, Kagan, C, Mannucci, R, Nicoletti, I, Teng, B & Denner, L 1999, 'Signaling pathway activated during apoptosis of the prostate cancer celt line LNCaP: Overexpression of caspase-7 as a new gene therapy strategy for prostate cancer', Cancer Research, vol. 59, no. 2, pp. 382-390.
Marcelli, Marco ; Cunningham, Glenn R. ; Walkup, Margaret ; He, Zening ; Sturgis, Lydia ; Kagan, Carolina ; Mannucci, Roberta ; Nicoletti, Ildo ; Teng, BaBie ; Denner, Larry. / Signaling pathway activated during apoptosis of the prostate cancer celt line LNCaP : Overexpression of caspase-7 as a new gene therapy strategy for prostate cancer. In: Cancer Research. 1999 ; Vol. 59, No. 2. pp. 382-390.
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AU - Walkup, Margaret

AU - He, Zening

AU - Sturgis, Lydia

AU - Kagan, Carolina

AU - Mannucci, Roberta

AU - Nicoletti, Ildo

AU - Teng, BaBie

AU - Denner, Larry

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