• The Effect of Silibinin on Apoptosis and PTEN in Human Breast Cancer Cell Line
  • Sanaz Ranapour,1 Nasrin Motamed,2,*
    1. Department of Cellular and Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran
    2. Department of Cellular and Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran


  • Introduction: Breast cancer is the most frequent malignancy in females. Amongst several naturally-occurring flavonoids, Silymarin is extracted from the seeds and fruits of milk thistle plant Silybum marianum which consists of main biologically active component as silibinin. It is known to inhibit cell proliferation, induce apoptosis, and curb angiogenesis. SB has demonstrated activity against many cancers, such as skin, liver, lung, bladder, and breast carcinomas. The PTEN gene acts as a tumor suppressor found in almost every tissue of the body. loss of function of the PTEN tumour suppressor is one of the most common events observed in many types of cancer.
  • Methods: 1. Cell lines and culture The human T47D breast cancer cell line was cultured in Medium DMEM. 2. MTT cell viability assay in T47D The human T47D breast cancer cell line was treated with different concentrations of silibinin (50- 250 μg/mL) for 24, 48 and 72 hours. The cytotoxic effect of silibinin on T47D viability was determined using Methyl-Thiazolyl-Tetrazolium (MTT) assay by IC50 determination. 3. Flow cytometric analysis for apoptotic Apoptosis was evaluated by Annexin V/propidium iodide staining. 4. Real-time PCR To assess the alterations of PTEN transcriptions, the real-time PCR reactions were performed using the Power SYBR-Green PCR Master Mix according to the manufacturer protocol. The relative gene expression levels were calculated using the 2−ΔΔCT method.
  • Results: IC50 concentrations of SB was144.6 % ± 9.41 for T47D cell after 48h treatment which demonstrated that SB cytotoxic activity on T47D was strong. Flow cytometry results illustrated that SB induced significant apoptosis cell death in T47D cell in comparison to untreated ones. mRNA expression levels demonstrate that SB significantly increased PTEN expression in T47D cell line, as compared to control groups.
  • Conclusion: Our results suggest that the apoptotic activity of SB in T47D cells. Our results have also revealed that SB can decelerate cancer cell progression and growth by targeting PTEN.
  • Keywords: Breast cancer, Silibinin,, Apoptosis, RT-PCR