Soft substrate induces apoptosis by the disturbance of Ca2+ homeostasis in renal epithelial LLC-PK1 cells

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Abstract

Different rigidities of adhesive collagen substrate affect cellular functions with unclear mechanisms. Here, we cultured a renal epithelial cell line (LLC-PK1) and a tumor cell line (HeLa)on substrates of different rigidities and compared the cell type-specific responses. The culture dish was coated with a very thin layer of collagen gel (control group) or overlaid with collagen gel (soft substrate). LLC-PK1 cells contracted as they grew on collagen gel and the apoptotic bodies obviously appeared with time. The protein levels of procaspase-12 and its downstream target procaspase-3 were decreased when LLC-PK1 cells cultured on collagen gel. μ-calpain was activated on collagen gel. Collage gel also induced the cleavage of α-spectrin which resulted in the disorganization of actin cytoskeleton. In contrast, there was no significant change in cytochrome c revelation, mitochondrial membrane potential, and the protein levels of procaspase-8 and procaspase-9. Moreover, soft substrate caused elevated cytosolic Ca , Ca2+ overload in ER and upregulation of capacitative calcium entry. Ca2+ chelator or channel blocker partially rescued the collagen-gel induced apoptosis by inhibiting μ-calpain activation. In contrast, for HeLa cells cultured either on collagen gel or on gel-coated dish, there was no significant change in positive Annexin V staining, no activation of procaspase-12 and no cleavage of μ-calpain. Thus, soft substrate induces apoptosis in LLC-PKI cells by the disturbance of Ca 2+ homeostasis.

Original languageEnglish
Pages (from-to)401-410
Number of pages10
JournalJournal of Cellular Physiology
Volume212
Issue number2
DOIs
Publication statusPublished - 2007 Aug 1

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

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