Induction of apoptosis in BPH stromal cells by adenoviral-mediated overexpression of caspase-7

Marco Marcelli, T. C. Shao, Xiaoying Li, Heather Yin, Michela Marani, Larry Denner, Babie Teng, Glenn R. Cunningham

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Purpose: We hypothesized that expression/activity of critical components of the apoptotic pathway can be used to induce apoptosis of a human prostate cell line derived from benign prostatic hyperplasia (BPH) tissue. Materials and Methods: We analyzed the apoptotic pathway in BPH cells treated with the powerful inducer of apoptosis, staurosporine (STS), and adenoviruses overexpressing caspase-3, -7, or the control gene lacZ. Results: Twelve hours post-STS, most BPH cells were floating in the culture medium, TUNEL staining was widespread, and DEVDase activity (the catalytic activity of type II caspases) was increased. The pan-caspase inhibitor, Z-VAD-FMK, prevented STS- induced apoptosis. Based on these observations, we performed immunoblot analysis for the three known group II caspases (that is caspase-2, -3 and - 7), but none of them was detected with three commercially available antibodies. Nevertheless, in view of the presence of increased DEVDase activity, we reasoned that a group II caspase must be a critical mediator of apoptosis in this model. If correct, we postulated that overexpression and activation of a type II caspase should cause apoptosis. To test this hypothesis, we coupled the cDNAs encoding caspase-3 and caspase-7 to adenoviral vectors and obtained constructs AvC3 and AvC7. Cells infected with AvC3 or AvC7 overexpressed the protein for caspase-3 or -7 within 24 to 48 hours. Caspase-3 overexpression did not cause apoptosis above that observed in cells receiving the control adenovirus expressing the lacZ cDNA (AvLac-Z). In contrast, caspase-7 overexpression induced massive apoptosis. BPH cells were then infected with increasing multiplicity of infection (MOI) of AvC7 and AvlacZ. A positive correlation was found between the amount of caspase-7 expressed and the level of DEVDase activity measured. AvC7 at MOIs of 25:1 and 50:1 induced apoptosis in about 50% of BPH cells at 72 hours post- infection. This effect was AvC7 specific, because the same MOIs of AvlacZ were not apoptogenic. Conclusions: Adenoviral-mediated overexpression of caspase-7 induces apoptosis of BPH-derived cells.

Original languageEnglish (US)
Pages (from-to)518-525
Number of pages8
JournalJournal of Urology
Volume164
Issue number2
StatePublished - Aug 2000
Externally publishedYes

Fingerprint

Caspase 7
Prostatic Hyperplasia
Stromal Cells
Apoptosis
Caspase 3
Caspases
Staurosporine
Adenoviridae
Complementary DNA
Caspase 2
Lac Operon
Caspase Inhibitors
In Situ Nick-End Labeling
Infection
Culture Media
Prostate
Staining and Labeling
Cell Line

Keywords

  • Apoptosis
  • Benign prostate hyperplasia
  • Caspases

ASJC Scopus subject areas

  • Urology

Cite this

Marcelli, M., Shao, T. C., Li, X., Yin, H., Marani, M., Denner, L., ... Cunningham, G. R. (2000). Induction of apoptosis in BPH stromal cells by adenoviral-mediated overexpression of caspase-7. Journal of Urology, 164(2), 518-525.

Induction of apoptosis in BPH stromal cells by adenoviral-mediated overexpression of caspase-7. / Marcelli, Marco; Shao, T. C.; Li, Xiaoying; Yin, Heather; Marani, Michela; Denner, Larry; Teng, Babie; Cunningham, Glenn R.

In: Journal of Urology, Vol. 164, No. 2, 08.2000, p. 518-525.

Research output: Contribution to journalArticle

Marcelli, M, Shao, TC, Li, X, Yin, H, Marani, M, Denner, L, Teng, B & Cunningham, GR 2000, 'Induction of apoptosis in BPH stromal cells by adenoviral-mediated overexpression of caspase-7', Journal of Urology, vol. 164, no. 2, pp. 518-525.
Marcelli, Marco ; Shao, T. C. ; Li, Xiaoying ; Yin, Heather ; Marani, Michela ; Denner, Larry ; Teng, Babie ; Cunningham, Glenn R. / Induction of apoptosis in BPH stromal cells by adenoviral-mediated overexpression of caspase-7. In: Journal of Urology. 2000 ; Vol. 164, No. 2. pp. 518-525.
@article{31e220f90b5c4e30b3cc3f74503a8b03,
title = "Induction of apoptosis in BPH stromal cells by adenoviral-mediated overexpression of caspase-7",
abstract = "Purpose: We hypothesized that expression/activity of critical components of the apoptotic pathway can be used to induce apoptosis of a human prostate cell line derived from benign prostatic hyperplasia (BPH) tissue. Materials and Methods: We analyzed the apoptotic pathway in BPH cells treated with the powerful inducer of apoptosis, staurosporine (STS), and adenoviruses overexpressing caspase-3, -7, or the control gene lacZ. Results: Twelve hours post-STS, most BPH cells were floating in the culture medium, TUNEL staining was widespread, and DEVDase activity (the catalytic activity of type II caspases) was increased. The pan-caspase inhibitor, Z-VAD-FMK, prevented STS- induced apoptosis. Based on these observations, we performed immunoblot analysis for the three known group II caspases (that is caspase-2, -3 and - 7), but none of them was detected with three commercially available antibodies. Nevertheless, in view of the presence of increased DEVDase activity, we reasoned that a group II caspase must be a critical mediator of apoptosis in this model. If correct, we postulated that overexpression and activation of a type II caspase should cause apoptosis. To test this hypothesis, we coupled the cDNAs encoding caspase-3 and caspase-7 to adenoviral vectors and obtained constructs AvC3 and AvC7. Cells infected with AvC3 or AvC7 overexpressed the protein for caspase-3 or -7 within 24 to 48 hours. Caspase-3 overexpression did not cause apoptosis above that observed in cells receiving the control adenovirus expressing the lacZ cDNA (AvLac-Z). In contrast, caspase-7 overexpression induced massive apoptosis. BPH cells were then infected with increasing multiplicity of infection (MOI) of AvC7 and AvlacZ. A positive correlation was found between the amount of caspase-7 expressed and the level of DEVDase activity measured. AvC7 at MOIs of 25:1 and 50:1 induced apoptosis in about 50{\%} of BPH cells at 72 hours post- infection. This effect was AvC7 specific, because the same MOIs of AvlacZ were not apoptogenic. Conclusions: Adenoviral-mediated overexpression of caspase-7 induces apoptosis of BPH-derived cells.",
keywords = "Apoptosis, Benign prostate hyperplasia, Caspases",
author = "Marco Marcelli and Shao, {T. C.} and Xiaoying Li and Heather Yin and Michela Marani and Larry Denner and Babie Teng and Cunningham, {Glenn R.}",
year = "2000",
month = "8",
language = "English (US)",
volume = "164",
pages = "518--525",
journal = "Journal of Urology",
issn = "0022-5347",
publisher = "Elsevier Inc.",
number = "2",

}

TY - JOUR

T1 - Induction of apoptosis in BPH stromal cells by adenoviral-mediated overexpression of caspase-7

AU - Marcelli, Marco

AU - Shao, T. C.

AU - Li, Xiaoying

AU - Yin, Heather

AU - Marani, Michela

AU - Denner, Larry

AU - Teng, Babie

AU - Cunningham, Glenn R.

PY - 2000/8

Y1 - 2000/8

N2 - Purpose: We hypothesized that expression/activity of critical components of the apoptotic pathway can be used to induce apoptosis of a human prostate cell line derived from benign prostatic hyperplasia (BPH) tissue. Materials and Methods: We analyzed the apoptotic pathway in BPH cells treated with the powerful inducer of apoptosis, staurosporine (STS), and adenoviruses overexpressing caspase-3, -7, or the control gene lacZ. Results: Twelve hours post-STS, most BPH cells were floating in the culture medium, TUNEL staining was widespread, and DEVDase activity (the catalytic activity of type II caspases) was increased. The pan-caspase inhibitor, Z-VAD-FMK, prevented STS- induced apoptosis. Based on these observations, we performed immunoblot analysis for the three known group II caspases (that is caspase-2, -3 and - 7), but none of them was detected with three commercially available antibodies. Nevertheless, in view of the presence of increased DEVDase activity, we reasoned that a group II caspase must be a critical mediator of apoptosis in this model. If correct, we postulated that overexpression and activation of a type II caspase should cause apoptosis. To test this hypothesis, we coupled the cDNAs encoding caspase-3 and caspase-7 to adenoviral vectors and obtained constructs AvC3 and AvC7. Cells infected with AvC3 or AvC7 overexpressed the protein for caspase-3 or -7 within 24 to 48 hours. Caspase-3 overexpression did not cause apoptosis above that observed in cells receiving the control adenovirus expressing the lacZ cDNA (AvLac-Z). In contrast, caspase-7 overexpression induced massive apoptosis. BPH cells were then infected with increasing multiplicity of infection (MOI) of AvC7 and AvlacZ. A positive correlation was found between the amount of caspase-7 expressed and the level of DEVDase activity measured. AvC7 at MOIs of 25:1 and 50:1 induced apoptosis in about 50% of BPH cells at 72 hours post- infection. This effect was AvC7 specific, because the same MOIs of AvlacZ were not apoptogenic. Conclusions: Adenoviral-mediated overexpression of caspase-7 induces apoptosis of BPH-derived cells.

AB - Purpose: We hypothesized that expression/activity of critical components of the apoptotic pathway can be used to induce apoptosis of a human prostate cell line derived from benign prostatic hyperplasia (BPH) tissue. Materials and Methods: We analyzed the apoptotic pathway in BPH cells treated with the powerful inducer of apoptosis, staurosporine (STS), and adenoviruses overexpressing caspase-3, -7, or the control gene lacZ. Results: Twelve hours post-STS, most BPH cells were floating in the culture medium, TUNEL staining was widespread, and DEVDase activity (the catalytic activity of type II caspases) was increased. The pan-caspase inhibitor, Z-VAD-FMK, prevented STS- induced apoptosis. Based on these observations, we performed immunoblot analysis for the three known group II caspases (that is caspase-2, -3 and - 7), but none of them was detected with three commercially available antibodies. Nevertheless, in view of the presence of increased DEVDase activity, we reasoned that a group II caspase must be a critical mediator of apoptosis in this model. If correct, we postulated that overexpression and activation of a type II caspase should cause apoptosis. To test this hypothesis, we coupled the cDNAs encoding caspase-3 and caspase-7 to adenoviral vectors and obtained constructs AvC3 and AvC7. Cells infected with AvC3 or AvC7 overexpressed the protein for caspase-3 or -7 within 24 to 48 hours. Caspase-3 overexpression did not cause apoptosis above that observed in cells receiving the control adenovirus expressing the lacZ cDNA (AvLac-Z). In contrast, caspase-7 overexpression induced massive apoptosis. BPH cells were then infected with increasing multiplicity of infection (MOI) of AvC7 and AvlacZ. A positive correlation was found between the amount of caspase-7 expressed and the level of DEVDase activity measured. AvC7 at MOIs of 25:1 and 50:1 induced apoptosis in about 50% of BPH cells at 72 hours post- infection. This effect was AvC7 specific, because the same MOIs of AvlacZ were not apoptogenic. Conclusions: Adenoviral-mediated overexpression of caspase-7 induces apoptosis of BPH-derived cells.

KW - Apoptosis

KW - Benign prostate hyperplasia

KW - Caspases

UR - http://www.scopus.com/inward/record.url?scp=0033927781&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033927781&partnerID=8YFLogxK

M3 - Article

VL - 164

SP - 518

EP - 525

JO - Journal of Urology

JF - Journal of Urology

SN - 0022-5347

IS - 2

ER -