Effect of cortisol on calpains in the C2C12 and 3T3-L1 cells

Pandurangan Muthuraman, Sambandam Ravikumar, Veerappan Muthuviveganandavel, Jongpil Kim

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

The present study was carried out to understand the effect of cortisol on calpain system in the C2C12 and 3T3-L1 adipocyte cells under co-culture system. Cells were co-cultured by using transwell inserts with a 0.4 μm porous membrane to separate C2C12 and 3T3-L1 preadipocyte cells. Each cell type was grown independently on the transwell plates. Following cell differentiation, inserts containing 3T3-L1 cells were transferred to C2C12 plates. Ten microgram per milliliter of cortisol was added to the medium. Following treatment for 3 days, the cells in the lower well were harvested for analysis. Calpains such as μ-calpain, m-calpain, and calpastatin were selected for the analysis. RT-PCR results indicated the significant increase in the mRNA expression of μ-calpain, m-calpain, and calpastatin. In addition, the confocal microscopical investigation indicated the cortisol treatment increases calpain expression in the C2C12 and 3T3-L1 cells. Taking all these together, cortisol treatment with co-culture system shows most reliable status of calpains expression in the cells, which is quite distinct from one-dimensional monocultured cells.

Original languageEnglish
Pages (from-to)3153-3162
Number of pages10
JournalApplied Biochemistry and Biotechnology
Volume172
Issue number6
DOIs
StatePublished - Mar 2014

Keywords

  • C2C12
  • Calpains
  • Protein
  • RT-PCR

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