Expression of Apoptotic Genes in MCF-7 Can-cer Cells after Induction with Human Adipose Stem Cells Conditioned Medium and Rosemary Extract

The effect of conditioned medium on apoptosis and invasion of MCF-7 is still debated. Carnosic acid, a component of rosemary extract, is also reported to have anti-cancer property. Therefore, we studied the occurrence of apoptosis through AIF-dependent pathway in MCF-7 cells treated with conditioned medium and rosemary extract by evaluating the expression of AIF, P53, Bcl-2 and Bax genes.

Method and Materials:

In this studty, the experimental groups were: cultured MCF7 cells in medium containing serum (control), cultured cells in conditioned medium containing serum (CM group), cultured cells in medium containing different concentrations of rosemary extract (50 and 100 μg / ml) for 24 and 48 hours (treatment groups). The cell proliferation rate was studied by MTT test. Gene expression was assessed by RT-PCR.

The survival rate of cells treated by conditioned medium and serum for 24 and 48 h was significantly decreased as compared to control. Meanwhile, there was a significant increase in the survival rate of MCF-7 cells cultured with rosemary 50 and 100 μg / ml containing serum for 48 h, compared to group treated with conditioned medium and serum. A significant increase in P53, AIF and Bax gene expression, and also a significant decrease in Bcl-2 gene expression were observed in cells treated with conditioned medium and serum for 24 and 48h, as compared to the control. Rosemary at the concentration of 100 µg/ml and serum for 48 h significantly decreased P53, AIF and Bcl-2 genes and rosemary at concentration of 50 µg/ml containing serum for 24 and 48 h significantly decreased Bax gene expression compared to conditioned medium and serum group.

The AIF-dependent apoptotic pathway was activated in MCF-7 cells after treatment with conditioned medium 24 and 48 h and rosemary 100 containing serum after 48 h.