Biochemical and morphological events during okadaic acid-induced apoptosis of Tsc2-null ERC-18 cell line

Todd Matthew Kolb, Seung H. Chang, Myrtle A. Davis

Research output: Contribution to journalArticle

Abstract

Several tumor suppressor genes have been shown to regulate cellular susceptibility to proliferation or apoptotic cell death. An essential first step in studies with the long-range goal of determining the effect of a tumor suppressor gene on cellular susceptibility to apoptosis is careful characterization of the cell's response to an apoptotic stimulus. The goals of this study were to characterize the apoptotic response of a tuberous sclerosis complex-2 (Tsc2) tumor suppressor gene-null cell line, to establish valid biochemical events that can be used as apoptosis markers, and to determine how these events correlate with apoptosis-specific morphologic changes. For characterization of apoptosis, we treated Tsc2-null renal epithelial tumor cells (ERC-18) with okadaic acid (OKA, 0.143.25 μM), and measured the biochemical and morphologic events during the apoptotic response. Electron microscopic and immunocytochemical evaluation showed an early loss of microvilli and a loss of vinculin and talin staining from focal adhesions within 1 hour. During the first 2 hours of treatment with 0.25 μM OKA, ERC-18 cells rounded and ∼50% detached from the culture vessel with minimal membrane bleb formation. Phosphatidylserine externalization, chromatin margination and fragmentation, cytochrome C release, and caspase-3 and -7 cleavage were evident at 6 hours. Maximal membrane bleb formation occurred between 6 and 10 hours. Cells progressed to secondary oncotic necrosis between 10 and 24 hours of OKA treatment. Almost all cells had an oncotic phenotype after 24 hours, and 17.5% lost cell membrane integrity. A small subpopulation (

Original languageEnglish (US)
Pages (from-to)235-246
Number of pages12
JournalToxicologic Pathology
Volume30
Issue number2
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Okadaic Acid
Tuberous Sclerosis
Tumors
Tumor Suppressor Genes
Cells
Apoptosis
Cell Line
Genes
Blister
Talin
Vinculin
Membranes
Caspase 7
Null Lymphocytes
Focal Adhesions
Phosphatidylserines
Cell death
Cell membranes
Cytochromes
Microvilli

Keywords

  • Apoptosis
  • Caspase
  • Morphology
  • Tsc2, okadaic acid
  • Tumor cell line

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Biochemical and morphological events during okadaic acid-induced apoptosis of Tsc2-null ERC-18 cell line. / Kolb, Todd Matthew; Chang, Seung H.; Davis, Myrtle A.

In: Toxicologic Pathology, Vol. 30, No. 2, 2002, p. 235-246.

Research output: Contribution to journalArticle

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