Evaluation of apoptosis and necrosis induced by statins using fluorescence-enhanced flow cytometry

J Pharm Biomed Anal. 2005 Sep 15;39(3-4):712-7. doi: 10.1016/j.jpba.2005.04.022.

Abstract

The purpose of this study was to evaluate the apoptosis and necrosis induced by five kinds of statins in IM-9 human lymphoblasts with fluorescence-enhanced flow cytometry using avidin-biotin complex. IM-9 human lymphoblasts (2 x 10(4) cells/cm2) were seeded into tissue culture plates and incubated with five kinds of statins. Statin-treated cells were first incubated with biotin-annexin V, followed by addition of avidin-FITC and propidium iodide, and then subjected to flow cytometry. The fluorescence intensity was enhanced using an avidin-biotin complex system, resulting in successful separate determination of the statin-induced apoptosis and necrosis by flow cytometry, which enabled us to quantitatively evaluate the statin-induced cell damage. Flow cytometric analysis results in the intensity of statin-induced apoptosis in IM-9 cells as follows: atorvastatin cerivastatin>fluvastatin simvastatin>pravastatin. The intensity of statin-induced necrosis in IM-9 cells was expressed as follows: atorvastatin cerivastatin>fluvastatin simvastatin>pravastatin. The total damage of IM-9 cells induced by five kinds of statins were expressed as the sum of both percentages of apoptosis and necrosis as follows: atorvastatin cerivastatin>fluvastatin simvastatin>pravastatin. Our studies show that fluorescence enhancement with avidin-biotin complex is useful for the identification and quantitation of annexin-positive apoptosis cells and thus, the fluorescence-enhanced flow cytometry was shown to be applicable for screening of statins as new anti-leukemia agents.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis*
  • Atorvastatin
  • Avidin / chemistry
  • Biotin / chemistry
  • Cell Line
  • Cell Separation / methods
  • Cells, Cultured
  • Drug Industry
  • Drug Screening Assays, Antitumor
  • Fatty Acids, Monounsaturated / pharmacology
  • Flow Cytometry / methods*
  • Fluorescein-5-isothiocyanate / pharmacology
  • Fluvastatin
  • Heptanoic Acids / pharmacology
  • Humans
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors / analysis*
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors / pharmacology
  • Indoles / pharmacology
  • Leukemia / drug therapy
  • Microscopy, Fluorescence / methods*
  • Necrosis*
  • Pharmaceutical Preparations / analysis
  • Pravastatin / pharmacology
  • Propidium / pharmacology
  • Pyridines / pharmacology
  • Pyrroles / pharmacology
  • Simvastatin / pharmacology
  • Time Factors

Substances

  • Fatty Acids, Monounsaturated
  • Heptanoic Acids
  • Hydroxymethylglutaryl-CoA Reductase Inhibitors
  • Indoles
  • Pharmaceutical Preparations
  • Pyridines
  • Pyrroles
  • Avidin
  • Propidium
  • Fluvastatin
  • Biotin
  • Atorvastatin
  • Simvastatin
  • cerivastatin
  • Fluorescein-5-isothiocyanate
  • Pravastatin