miR-24-2 targets Akt and inhibits cell survival, EMT, and tumour growth in triple-negative breast cancer.

BACKGROUND

Breast cancer is one of the most common types of cancer in the world, and triple-negative breast cancer (TNBC) is an extremely aggressive subtype. This study aimed to investigate the efficacy and mechanisms of miR-24-2 against TNBC.

METHODS

MDA-MB-231 (TNBC), HEK293T and HEK293 cells were used for various assays including cell viability and proliferation, cell migration, and invasion. RT-PCR, immunoblotting, acridine orange-ethidium bromide and DCFDA assays, tumour xenograft, H&E and IHC methods were employed in the study.

RESULTS

miR-24-2 overexpression induced cell death and inhibited clonogenic potential. It also reduced the expression of Slug, Twist1, and Vimentin and inhibited cell invasion and migration. miR-24-2 enhanced apoptosis, ROS production, and expression of pH2A.X (S139) while decreased Chk1, Chk2, and Rad51. miR-24-2 inhibited survival signaling by downregulating Akt, Erk1/2, Bcl-2, and cytochrome c (mitochondrial), and enhancing Bax, cytochrome c (cytosolic), and cleaved-caspase 3. miR-24-2 binds to the coding sequence (CDS) of Akt which was validated at mRNA and protein expression levels. Further, miR-24-2 strongly reduced TNBC tumour volume (58 %, p < 0.01) and weight (54 %, p < 0.001) during 20 days. Tumour immunohistochemistry showed that miR-24-2 decreased proliferation and increased apoptosis. Moreover, increased Bax/Bcl-2 ratio and decreased expression of Akt, PCNA, Chk1, Chk2, and Rad51 by miR-24-2 were found in tumours, validating the findings of in vitro and in vivo tumour models.

CONCLUSION

Overall, this study finds that miR-24-2 directly targets Akt and plays a significant role in TNBC as a key mechanism for its growth, survival, and progression.