Effect of cortisol on calpains in the C2C12 and 3T3-L1 cells.
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.
Department of Biomedical Engineering, Dongguk University, Seoul, South Korea, frenzram80@gmail.com.
Pandurangan
P
Muthuraman
Sambandam
S
Ravikumar
Veerappan
V
Muthuviveganandavel
Jongpil
J
Kim
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D002154:Calpain / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D018920:Coculture Techniques; D015194:Diffusion Chambers, Culture; D015870:Gene Expression / Q000187:drug effects*; D006854:Hydrocortisone / Q000494:pharmacology*; D051379:Mice; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism
D000893:Anti-Inflammatory Agents; D002135:Calcium-Binding Proteins; D012333:RNA, Messenger; C033668:calpastatin; D002154:Calpain; C449000:m-calpain; C449002:mu-calpain; D006854:Hydrocortisone