Paclitaxel-induced apoptosis in non-small cell lung cancer cell lines is associated with increased caspase-3 activity

J Thorac Cardiovasc Surg. 2000 Apr;119(4 Pt 1):795-803. doi: 10.1016/S0022-5223(00)70016-X.

Abstract

Objective: Our objective was to determine whether paclitaxel-induced apoptosis in human lung cancer cells is Fas dependent.

Methods: Human lung cancer cell lines were evaluated for morphologic evidence of apoptosis, DNA fragmentation (TUNEL positivity), and caspase-3 activation after paclitaxel treatment. Human lung adenocarcinoma, squamous cell carcinoma, undifferentiated lung carcinoma, and bronchoalveolar carcinoma cell lines were each cultured in 10 micromol/L paclitaxel.

Results: After 24 hours of culture in paclitaxel, a 22% to 69% increase in the number of apoptotic cells was evident by means of methylene blue-azure A-eosin staining with characteristic blebbing and nuclear condensation. TUNEL assay also confirmed an increase of 19.9% to 73.0% of cells with nuclear fragmentation. Caspase-3 activity, assayed by Z-DEVD cleavage, increased from 20% to 215% (P <.05). ZB4, an antagonistic anti-Fas antibody, did not block paclitaxel induction of caspase-3 activity (155.8 vs 165.8 U, not significant). Apoptotic morphologic changes were inhibited in cells cultured in the presence of paclitaxel and Ac-DEVD-CHO, a caspase-3 inhibitor.

Conclusions: Paclitaxel induces apoptosis in lung cancer cell lines, as assessed by a consistent increase in caspase-3 activity, DNA laddering, and characteristic morphologic changes. Paclitaxel-induced apoptosis in human lung cancer cells is associated with caspase-3 activation but is not Fas dependent.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Apoptosis / drug effects*
  • Carcinoma, Non-Small-Cell Lung / chemistry
  • Carcinoma, Non-Small-Cell Lung / enzymology
  • Carcinoma, Non-Small-Cell Lung / pathology*
  • Caspase 3
  • Caspase Inhibitors
  • Caspases / metabolism*
  • DNA Fragmentation
  • Humans
  • In Situ Nick-End Labeling
  • Lung Neoplasms / chemistry
  • Lung Neoplasms / enzymology
  • Lung Neoplasms / pathology*
  • Oligopeptides / pharmacology
  • Paclitaxel / pharmacology*
  • Tumor Cells, Cultured
  • fas Receptor / analysis

Substances

  • Antineoplastic Agents, Phytogenic
  • Caspase Inhibitors
  • Oligopeptides
  • acetyl-aspartyl-glutamyl-valyl-aspartal
  • fas Receptor
  • CASP3 protein, human
  • Caspase 3
  • Caspases
  • Paclitaxel