Effects of Moringa oleifera Lam. extracellular vesicles on endogenous miRNAs involved in tumorigenesis in HeLa cell line.

Introduction: Cervical cancer (CC) is one of the most frequent cancers in women worldwide. The epithelial mesenchymal transition (EMT) and the extracellular release of TGF-β are phenomena associated with different tumorigenic processes. Specific human microRNAs (h-miRs) involved in these processes have been identified, becoming important diagnostic and prognostic markers, and potential therapeutic targets. As reported in the literature and in our previous studies, plant Extracellular Vesicles (EVs) modulated host’s gene expression in pathological conditions, such as cancer. In the present study, the effects of EVs from Moringa oleifera Lam (MOES-EVs) on the modulation of tumour associated h-miRs profile in CCderived HeLa cells were investigated. Methods: EVs were purified from MOES and characterized by CytoFLEX flow cytometer. The phenols and f lavonoids content were analysed by HPLC-DAD and quantified by spectrophotometric assays. The ability of MOES-EVs to enter HeLa cells was assessed by SYTO RNA staining analysed by CytoFLEX and inverted microscope. Eighty-four h-miRs were analysed in HeLa cells treated with MOES-EVs using the Human Cancer Pathway PCR Array. The effect of MOES-EVs treatment on cell migration and TGF-β expression were analysed by scratch test and ELISA respectively. Results: The HeLa cells treated with MOES-EVs showed a modified tumour suppressor and oncogenic h-miRs profile that affected cell proliferation, apoptosis and migration. Among the 84 h-miRs analysed, 19 were unexpressed in both treated and untreated HeLa, 47 increased their expression under treatment, 4 decreased and 14 were unchanged. The treatment with MOESEVs modulated 60,7% of h-miRs analysed in this study, upregulating 34,5% of tumour suppressors h-miRs and down-regulating 4,7% of oncomiRs. Finally, our data confirmed the ability of MOES-EVs to modulate migration reducing the TGF-β protein expression and significantly inhibiting cell motility in an in vitro model. Conclusion: Our results pave the way for the development of new potential adjuvants approach for CC treatment based on plant EVs modulating endogenous miRs involved in tumorigenesis.