Paclitaxel induces apoptosis in Saos-2 cells with CD95L upregulation and Bcl-2 phosphorylation

Bruna Pucci, Lorenza Bellincampi, Marco Tafani, Valeria Masciullo, Gerry Melino, Antonio Giordano

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

We examined the effect of paclitaxel on human osteoblastic cells Saos-2 to determine if paclitaxel can affect, proliferation and apoptosis. We used a p53-negative cell line in order to mimic the loss of function frequently observed at the clinical level. Paclitaxel induced cell death in a dose- and time-dependent manner. Marked nuclear condensation and fragmentation of chromatin were observed by Hoechst 33258 stain, DNA ladder formation, electron microscopy, and flow cytometry at concentrations as low as 100 nM, a concentration which can be achieved by infusion in human plasma. At 100 nM, paclitaxel induced a G2 arrest at 8 h of treatment. The cells then continued to accumulate in G2 until 72 h when the percentage of apoptotic events reached 54%. At the molecular level, Bcl-2 protein was phosphorylated at 16 h and PARP protein was cleaved, indicating the activation of caspase-3-like proteases. Caspase inhibitors Z-VAD-FMK and Z-DEVD-FMK rescued Saos-2 cells from paclitaxel-induced apoptosis. CD95 expression was constantly high, while CD95L showed a threefold increase in expression. This suggests that, following the G2 arrest, apoptosis is induced through the CD95/CD95L system.

Original languageEnglish
Pages (from-to)134-143
Number of pages10
JournalExperimental Cell Research
Volume252
Issue number1
DOIs
Publication statusPublished - Oct 10 1999

Fingerprint

Fas Ligand Protein
Paclitaxel
Up-Regulation
Phosphorylation
Apoptosis
Bisbenzimidazole
Caspase Inhibitors
Caspase 3
Chromatin
Electron Microscopy
Flow Cytometry
Proteins
Peptide Hydrolases
Cell Death
Coloring Agents
Cell Line
DNA

Keywords

  • Apoptosis
  • CD95°Bcl-2
  • Osteosarcoma
  • Paclitaxel

ASJC Scopus subject areas

  • Cell Biology

Cite this

Paclitaxel induces apoptosis in Saos-2 cells with CD95L upregulation and Bcl-2 phosphorylation. / Pucci, Bruna; Bellincampi, Lorenza; Tafani, Marco; Masciullo, Valeria; Melino, Gerry; Giordano, Antonio.

In: Experimental Cell Research, Vol. 252, No. 1, 10.10.1999, p. 134-143.

Research output: Contribution to journalArticle

Pucci, Bruna ; Bellincampi, Lorenza ; Tafani, Marco ; Masciullo, Valeria ; Melino, Gerry ; Giordano, Antonio. / Paclitaxel induces apoptosis in Saos-2 cells with CD95L upregulation and Bcl-2 phosphorylation. In: Experimental Cell Research. 1999 ; Vol. 252, No. 1. pp. 134-143.
@article{4a483f77e1c444d7908e917fe32f0078,
title = "Paclitaxel induces apoptosis in Saos-2 cells with CD95L upregulation and Bcl-2 phosphorylation",
abstract = "We examined the effect of paclitaxel on human osteoblastic cells Saos-2 to determine if paclitaxel can affect, proliferation and apoptosis. We used a p53-negative cell line in order to mimic the loss of function frequently observed at the clinical level. Paclitaxel induced cell death in a dose- and time-dependent manner. Marked nuclear condensation and fragmentation of chromatin were observed by Hoechst 33258 stain, DNA ladder formation, electron microscopy, and flow cytometry at concentrations as low as 100 nM, a concentration which can be achieved by infusion in human plasma. At 100 nM, paclitaxel induced a G2 arrest at 8 h of treatment. The cells then continued to accumulate in G2 until 72 h when the percentage of apoptotic events reached 54{\%}. At the molecular level, Bcl-2 protein was phosphorylated at 16 h and PARP protein was cleaved, indicating the activation of caspase-3-like proteases. Caspase inhibitors Z-VAD-FMK and Z-DEVD-FMK rescued Saos-2 cells from paclitaxel-induced apoptosis. CD95 expression was constantly high, while CD95L showed a threefold increase in expression. This suggests that, following the G2 arrest, apoptosis is induced through the CD95/CD95L system.",
keywords = "Apoptosis, CD95°Bcl-2, Osteosarcoma, Paclitaxel",
author = "Bruna Pucci and Lorenza Bellincampi and Marco Tafani and Valeria Masciullo and Gerry Melino and Antonio Giordano",
year = "1999",
month = "10",
day = "10",
doi = "10.1006/excr.1999.4591",
language = "English",
volume = "252",
pages = "134--143",
journal = "Experimental Cell Research",
issn = "0014-4827",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Paclitaxel induces apoptosis in Saos-2 cells with CD95L upregulation and Bcl-2 phosphorylation

AU - Pucci, Bruna

AU - Bellincampi, Lorenza

AU - Tafani, Marco

AU - Masciullo, Valeria

AU - Melino, Gerry

AU - Giordano, Antonio

PY - 1999/10/10

Y1 - 1999/10/10

N2 - We examined the effect of paclitaxel on human osteoblastic cells Saos-2 to determine if paclitaxel can affect, proliferation and apoptosis. We used a p53-negative cell line in order to mimic the loss of function frequently observed at the clinical level. Paclitaxel induced cell death in a dose- and time-dependent manner. Marked nuclear condensation and fragmentation of chromatin were observed by Hoechst 33258 stain, DNA ladder formation, electron microscopy, and flow cytometry at concentrations as low as 100 nM, a concentration which can be achieved by infusion in human plasma. At 100 nM, paclitaxel induced a G2 arrest at 8 h of treatment. The cells then continued to accumulate in G2 until 72 h when the percentage of apoptotic events reached 54%. At the molecular level, Bcl-2 protein was phosphorylated at 16 h and PARP protein was cleaved, indicating the activation of caspase-3-like proteases. Caspase inhibitors Z-VAD-FMK and Z-DEVD-FMK rescued Saos-2 cells from paclitaxel-induced apoptosis. CD95 expression was constantly high, while CD95L showed a threefold increase in expression. This suggests that, following the G2 arrest, apoptosis is induced through the CD95/CD95L system.

AB - We examined the effect of paclitaxel on human osteoblastic cells Saos-2 to determine if paclitaxel can affect, proliferation and apoptosis. We used a p53-negative cell line in order to mimic the loss of function frequently observed at the clinical level. Paclitaxel induced cell death in a dose- and time-dependent manner. Marked nuclear condensation and fragmentation of chromatin were observed by Hoechst 33258 stain, DNA ladder formation, electron microscopy, and flow cytometry at concentrations as low as 100 nM, a concentration which can be achieved by infusion in human plasma. At 100 nM, paclitaxel induced a G2 arrest at 8 h of treatment. The cells then continued to accumulate in G2 until 72 h when the percentage of apoptotic events reached 54%. At the molecular level, Bcl-2 protein was phosphorylated at 16 h and PARP protein was cleaved, indicating the activation of caspase-3-like proteases. Caspase inhibitors Z-VAD-FMK and Z-DEVD-FMK rescued Saos-2 cells from paclitaxel-induced apoptosis. CD95 expression was constantly high, while CD95L showed a threefold increase in expression. This suggests that, following the G2 arrest, apoptosis is induced through the CD95/CD95L system.

KW - Apoptosis

KW - CD95°Bcl-2

KW - Osteosarcoma

KW - Paclitaxel

UR - http://www.scopus.com/inward/record.url?scp=0033544005&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033544005&partnerID=8YFLogxK

U2 - 10.1006/excr.1999.4591

DO - 10.1006/excr.1999.4591

M3 - Article

VL - 252

SP - 134

EP - 143

JO - Experimental Cell Research

JF - Experimental Cell Research

SN - 0014-4827

IS - 1

ER -