Cytochrome c release and endoplasmic reticulum stress are involved in caspase-dependent apoptosis induced by G418

Q. H. Jin, B. Zhao, X. J. Zhang

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

G418 is used extensively in transfection experiments to select eukaryotic cells that have acquired neomycin resistance genes, but the mechanism is still elusive. To investigate this, we treated normal rat kidney cells with G418 for 3 days and found that the cells presented typical apoptotic features such as cell shrinkage, nuclear fragmentation, and caspase-3 activation. However, there was no low-molecular DNA ladder. The pan caspase inhibitor z-VAD-fmk completely inhibited this type of apoptosis, suggesting a caspase-dependent mechanism. Caspase cascades in apoptosis induced by G418 were initiated by at least two pathways: the release of cytochrome c from mitochondria, which was observed under confocal microscopy, and endoplasmic reticulum stress, demonstrated by the increase in Ca2+ concentration and the cleavage of m-calpain and procaspase-12. Both pathways activated caspase-9. Inhibition of caspase-9 activity by z-LEHD-fmk prevented most of the cells from apoptosis, and E-64d, an inhibitor of calpain accentuated this block. The cleavage of casapse-9 and caspase-12 was blocked only by simultaneous application of z-VAD-fmk and E-64d, but not by either alone. E-64d did not prevent the release of cytochrome c. These results indicated that these two pathways were independent of each other.

Original languageEnglish (US)
Pages (from-to)1816-1825
Number of pages10
JournalCellular and Molecular Life Sciences
Volume61
Issue number14
StatePublished - Jul 1 2004

Fingerprint

Endoplasmic Reticulum Stress
Caspases
Cytochromes c
Apoptosis
Caspase 9
Caspase 12
Caspase Inhibitors
Neomycin
Eukaryotic Cells
Confocal Microscopy
Caspase 3
Transfection
Mitochondria
Kidney
antibiotic G 418
DNA
Genes

Keywords

  • Apoptosis
  • Cytochrome c
  • Endoplasmic reticulum stress
  • G418

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Cellular and Molecular Neuroscience
  • Cell Biology

Cite this

Cytochrome c release and endoplasmic reticulum stress are involved in caspase-dependent apoptosis induced by G418. / Jin, Q. H.; Zhao, B.; Zhang, X. J.

In: Cellular and Molecular Life Sciences, Vol. 61, No. 14, 01.07.2004, p. 1816-1825.

Research output: Contribution to journalArticle

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