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, Ba Bie Teng, Larry Denner

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115 Scopus citations

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

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ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Marcelli, M., Cunningham, G. R., Walkup, M., He, Z., Sturgis, L., Kagan, C., Mannucci, R., Nicoletti, I., Teng, B. 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, 59(2), 382-390.