Distinct Oxidative Stress Adaptations Driven by the Overexpression of miR-526b, miR-655, and COX-2 in Breast Cancer

Oxidative stress has a dual role in breast cancer, promoting growth at moderate levels while causing cell death at higher levels, such as during therapeutic interventions that increase reactive oxygen species production. Oncogenic microRNAs miR-526b and miR-655 promote aggressive cancer traits-such as proliferation, migration, invasion, hypoxia response, cancer stem cell properties, and metastasis-via COX-2/EP4/PI3K pathways. These miRNAs and oxidative stress appear to engage in a self-amplifying loop, where miRNA overexpression increases ROS levels, and moderate oxidative stress, in turn, enhances miRNA expression-although the mechanisms are not yet fully understood. This study investigates how overexpressing miR-526b, miR-655, and COX-2 influences breast cancer cell responses to oxidative stress induced by H₂O₂. We examined cell viability, DNA damage, and transcriptomic changes in MCF7, MCF7-miR526b, MCF7-miR655, and MCF7-COX2 cell lines. Overexpression of COX-2 provided the most significant protection against oxidative stress, decreasing apoptosis and promoting cell cycle progression. Cells with miR-526b and miR-655 exhibited distinct yet overlapping stress responses, including decreased expression of DNA damage markers and alterations in p53 signaling. RNA-sequencing and network analyses identified hub genes involved in redox balance, immune, and metabolic pathways, which may have clinical significance (<i>OAS2</i>, <i>TNF</i>, <i>CACNA1C</i>, <i>CALML5</i>). Overall, these findings suggest that miR-526b, miR-655, and COX-2 play novel roles in promoting resistance to oxidative stress through transcriptional reprogramming in breast cancer; the identified markers could serve as potential biomarkers or therapeutic targets.