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  • Investigating the Effects of miR-204-Enriched Mesenchymal Stem Cell-Derived Exosomes on TGF-β1-Induced EMT in 4T1 Breast Cancer Cells

  • Maryam Etesami ,1,*
    1. Department of Genetics, Faculty of Basic Sciences, Central Tehran Branch, Islamic Azad University, Tehran, Iran


  • Introduction: Breast cancer is a prevalent disease among females throughout the world. Epithelial-Mesenchymal Transition (EMT) is an essential procedure that plays a key role in cancer development, invasion, and metastasis. Transforming Growth Factor-Beta 1 (TGF-β1) is a significant inducer of EMT and is capable of inducing a malignant phenotype in breast cancer cells. Micro RNAs such as miR-204 have already been linked to the regulation of tumor progression and EMT. Mesenchymal Stem Cell-Derived Exosomes are novel vectors for bioactive materials and can influence cancer-related signaling pathways. The present study aims to determine the impact of miR-204-containing Mesenchymal Stem Cell-Derived Exosomes on TGF-β1-induced EMT in 4T1 breast cancer cells.
  • Methods: Mesenchymal stem cells (MSCs) were prepared and cultivated from the Iranian Pasteur Institute miR-204 mimic was transmitted to cells. The cellular RNA was extracted and the CDNA was synthesized and was conducted to investigate the miR-204 RT-QCR expression. The exosome was extracted from transformation cells and control. Exosomes' validity tests were performed. The size of the exosomes was evaluated by the DLS device. Exosome was investigated using an electron microscope or TEM. Exosome and TGF-1 treatment were performed. The MTT test was conducted to determine the toxicity of cells following treatment with different concentrations of TGF-1 in the presence and absence of exosome. Expression of EMT-related transcription factors in the cells treated with TGF-1 and Exosome Extraction of CDNA and RT-PCR Synthesis were performed to investigate the expression of N-Cadherin, E-Cadherin and vimentin genes.
  • Results: According to the analyzes, the spherical morphological exosomes with an approximate diameter of 50-50 nm were observed by an electron microscope or TEM. Transformation of mesenchymal stem cells with miR-204 mimic led to more meaningful expression of miR-204 in cells 4T1s. Real-Time PCR results showed that miR-204 with higher levels in derivatives of mesenchymal stem cells with miR-204 mimic Oligonucleotides were transmitted (**p< 0.01). Separate TGF-β also had increasing effects on the expression of N-Cadherin and Vimentin genes. While the expression of the E-Cadherin epithelial marker was reduced after the treatment. These results are presented compared to the control group (non -transfect stem cell exosomes). The results of MTT Assay showed that half of the cells were alive at a concentration of 20 nm of TGF-β at a concentration of 20 nm, so the IC50 was set at 20 nm.
  • Conclusion: Based on these results, it can be speculated that miR-204-loaded MSC exosomes might affect the EMT-associated molecular changes caused by TGF-β1 in 4T1 breast cancer cells. The changes in E-cadherin, N-cadherin, and vimentin expression levels suggest the possible involvement of miR-204-loaded exosomes in EMT regulation. Future investigations are needed to identify the mechanism behind this phenomenon and understand the clinical significance of this treatment strategy for breast cancer.
  • Keywords: Breast Cancer, Exosomes, MicroRNA, TGF-β, miR-204 mimic, Messania Stem Cells

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