Title

Effect of Cortisol on Calpains in the C2C12 and 3T3-L1 Cells.

Authors

Muthuraman, Pandurangan; Ravikumar, Sambandam; Muthuviveganandavel, Veerappan; Kim, Jongpil

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.

Subjects

HYDROCORTISONE regulation; CALPAIN; CYSTEINE proteinases; CELL differentiation; CELL culture; CELL membranes

Publication

Applied Biochemistry & Biotechnology, 2014, Vol 172, Issue 6, p3153

ISSN

0273-2289

Publication type

Academic Journal

DOI

10.1007/s12010-014-0753-1