MiR-144 as a novel biomarker in breast cancer diagnosis and treatment

Abstract

Exosomes naturally carry the biomolecules in the body; they perform this task efficiently without compromising the immune system and by breaking through all the biological barriers, so they can be the best choice for designing and introducing a drug and gene transfer system. Extraction of the exosomes from the cell culture medium was performed by precipitation with an Exoquick kit solution. Nanoparticle specificity analysis was performed using scanning electron microscopy and dynamic light scattering. Trizol reagent (Invitrogen) was used for RNA extraction. Single-strand cDNA synthesis was performed from the miRNA and RT-PCR. Data were analyzed using a threshold cycle comparative method and cell cycle analysis using flow cytometry. Exosomes containing miR-144 can dramatically decrease the expression level of crucial TGF-β pathway genes, SMAD4 and TGF-βR2, in breast cancer cells. Botulinum toxin A inhibits cancer cell growth by inhibiting the TGF-β pathway. The simultaneous combination of engineered exosomes containing miR-144 and bacterial botulinum toxin A has increased effects on inhibiting the TGF-β signaling pathway. It causes cell cycle arrest in breast cancer cells. The present study's findings showed that overexpression of miR-144 in breast tumor cells results in the packaging of miRNA in exosomes derived from these cells. As a result, the exosomal platform for nucleic acid transfer to the cell appears to be an effective transducer for gene transfer to the cell. It could be used as a suitable adjunct to cancer therapeutic studies. Keywords: Breast cancer, botulinum toxin A, exosome, miRNA, biomarker.