Effects of calcitriol on the cell cycle of rhabdomyosarcoma cells

Abstract

Rhabdomyosarcoma (RMS) is a type of cancer of skeletal muscle. Calcitriol is the active form of vitamin D3, also recognised as a steroid hormone called 1α, 25-dihydroxy vitamin D3 (1,25D). We previously reported that 1,25D promoted cell proliferation and differentiation in non-cancerous skeletal muscle cells C2C12. The aim of this work is to evaluate some of the events triggered by 1,25D in RD cells, a human RMS cell line. In this work we reported that RD cells expressed vitamin D receptor (VDR) and treatment with 1,25D reduced VDR expression at 72 h. At the same time an acute decrease in viable cells as well as in cells in S-phase of cell cycle was also observed. Furthermore, up-regulation of p15INK4b was accompanied in a timely manner by down-regulation of cyclin D3, p21Waf1/Cip1 and myogenin protein levels. Simultaneously, 1,25D induced early apoptosis markers such as cyclin D1 and CDK4, and the disruption of the mitochondrial network together with a redistribution of mitochondria around the nucleus. Finally, 1,25D induced changes in the plasma membrane of RD cells associated with early and late apoptosis at 72 h, as determined by flow cytometry. Taken together, these results determine that treatment with 1,25D for 72 h triggers apoptosis in RD cells.