The Synergic Effects of Crocus Sativus L. and Low Frequency Electromagnetic Field on VEGFR2 Gene Expression in Human Breast Cancer Cells

Abstract

Background: Angiogenesis, which is required for embryonic development and many physiological events, plays crucial role in many pathological conditions such as tumor growth and metastasis. Recent studies indicate anticancer and antitumor properties of saffron against human cancers. Many processes are affected by Electromagnetic Field (EMF) and its effect on proliferation and gene expression were examined. In this experimental study, the synergic effects of saffron and EMF on VEGFR2 gene expression in MCF7 cells were investigated.

Methods: Saffron was extracted using freeze dryer. MCF7 cells were grown in RPMI 1640 medium supplemented with 10% FBS and incubated at 37°C with 5% CO2. After 24 hr cells were treated with saffron extract at concentrations of 100, 200, 400 and 800 µg/ml. Forty eight hr after treatment all flasks were exposed with EMF (50 Hz, 0.004 T). Then total RNA was extracted and cDNA was synthetized using specific primer. Synthetized products were analyzed by Real Time PCR to determine expression level of VEGFR2. Data were analyzed by SPSS (ANOVA & Tukey).

Results: Critical inhibitory effect on VEGFR2 gene expression was 20% at 400 µg/ml. Synergic use of EMF and saffron extract showed most reduction (38%) at 100 µg/ml. On the other hand synergic use of 200, 400 and 800 µg/ml saffron aqua extract and EMF decline noticeably the VEGFR2 level of gene expression to 29, 35 and 36%, respectively. EMF itself also reduced VEGFR2 up to 25% in comparison with control group which is remarkable at p < 0.001.

Conclusion: Results indicate a decrease in the expression of vascular endothelial growth factor receptor in the treated samples with saffron extract compared to control. This reduction in VEGFR2 level induced by synergic treatment of saffron and EMF which reveals induction of inhibitory effects of saffron on angiogenesis and could be also considered as a promising chemotherapeutic agent in breast cancer treatment.

Keywords: Angiogenesis; Cancer; MCF7 cells; Saffron; VEGFR2.

Figure 1

Housekeeping gene (β-Actin) expression in comparison with target gene (VEGFR₂)

Figure 2

VEGFR₂ gene expression in samples treated with saffron aqueous extract in different concentrations and samples treated with saffron aqueous extract µg/ml exposed to electromagnetic field for one hr. * p < 0.05, **p < 0.001

Figure 3

CT scaling down by using standard dilution samples with quantities of 100, 10, 0.1, and 0.01.