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  • Evaluation of miR-622 and OCT4 Expression in Breast Cancer Cells Treated with Extracellular Vesicles Derived from Stromal Cells

  • Fateme Hanifi,1 Mina ramezani ,2,*
    1. Department of Biology, CT.C., Islamic Azad University, Tehran, Iran.
    2. Department of Biology, CT.C., Islamic Azad University, Tehran, Iran. an.


  • Introduction: Breast cancer is one of the most common cancers in women and remains a major cause of cancer-related mortality worldwide. The tumor microenvironment plays an important role in regulating cancer cell behavior and disease progression. Tumor stromal cells can release extracellular vesicles (EVs) and modulate intercellular communication within the tumor microenvironment through the transfer of various bioactive molecules. MicroRNAs are important regulatory molecules involved in cancer-related cellular processes. miR-622 has been implicated in the regulation of cancer-associated pathways, while OCT4 is a key transcription factor involved in maintaining stem cell characteristics. Dysregulated OCT4 expression in cancer cells may be associated with stem-like properties and tumor progression. Given the potential role of stromal-derived EVs in modulating tumor cell characteristics, this study aimed to investigate the effect of EVs derived from breast tumor stromal cells on miR-622 and OCT4 expression in MDA-MB-231 breast cancer cells.
  • Methods: Breast tumor stromal cells obtained from primary culture were characterized based on their morphological features, including spindle-shaped morphology and cell adhesion, and were maintained under standard cell culture conditions. The MDA-MB-231 breast cancer cell line was also cultured and maintained under standard conditions. To investigate the potential effects of stromal-derived EVs on breast cancer cells, EVs were isolated from the culture supernatant of breast tumor stromal cells using the ExoQuick kit and characterized by dynamic light scattering (DLS). The appropriate dose was determined by MTT assay. MDA-MB-231 cells were subsequently treated with the isolated EVs. Following treatment, cellular RNA was extracted, and its quality was assessed. The extracted RNA was then reverse-transcribed into cDNA, and the expression levels of miR-622 and OCT4 were evaluated using real-time PCR. Expression levels were normalized to the appropriate internal control and compared between the treatment and control groups. Data were expressed as mean ± standard deviation and analyzed using the unpaired Student’s t-test.
  • Results: Treatment of MDA-MB-231 cells with EVs (30 µg/mL) derived from breast tumor stromal cells resulted in a significant decrease in miR-622 expression compared with the control group (*p<0.05). In contrast, a significant increase in OCT4 expression was observed in EV-treated MDA-MB-231 cells compared with the untreated control group (*p<0.05).
  • Conclusion: The findings indicate that EVs derived from breast tumor stromal cells with concentration can modulate miR-622 and OCT4 expression in MDA-MB-231 breast cancer cells. The decreased miR-622 expression accompanied by increased OCT4 expression following stromal-derived EV treatment may reflect alterations in molecular pathways associated with stem-like and malignant characteristics of breast cancer cells. These findings highlight the potential role of EV-mediated communication between stromal and tumor cells and provide a basis for further investigation of the molecular mechanisms underlying interactions between the tumor microenvironment and breast cancer cells.
  • Keywords: Breast cancer, extracellular vesicles , tumor stromal cells, miR-622, OCT4, MDA-MB-231

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