4-chloro-1,2-phenylenediamine Induces Apoptosis in Mardin-Darby canine kidney cells via activation of caspases

Leong Chee Onn, Chen Ssu Ching, Tiong Yee Lian, Loh Veng Foon, Ng Chew Hee, Chye Soi Moi

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4-Chloro-1,2-phenylenediamine (4-Cl-o-PD) is a halogenated aromatic diamine that was used as a precursor for manufacturing permanent hair dyes. Despite its well-documented mutagenic and carcinogenic effects in a number of in vitro and in vivo models, its cytotoxicity and mode of action have not received similar attention. Here, we investigated the effect of 4-Cl-o-PD on Mardin-Darby canine kidney cells. It induced apoptosis and the evidence suggests its initiation by reactive oxygen species (ROS). The results of various assays used show a dose-dependent (i) decrease in cell viability, (ii) increase in cells at sub-G1 phase and the G0/G1 phase arrested in cell cycle, (iii) increase in intracellular ROS accompanied by depletion of glutathione, and (iv) that apoptotic cell death probably involves activation of both intrinsic and extrinsic pathways.

Original languageEnglish
Pages (from-to)655-664
Number of pages10
JournalEnvironmental Toxicology
Issue number6
StatePublished - Jun 2014


  • 4-chloro-1,2-phenylenediamine
  • Apoptosis
  • Caspase
  • Mardin-Darby canine kidney cells
  • Reactive oxygen species


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