SIRT3 silencing sensitizes breast cancer cells to cytotoxic treatments through an increment in ROS production

Abstract

SIRT3, the major deacetylase in mitochondria, plays a crucial role modulating ROS production and scavenging by regulating key proteins implicated in mitochondrial turnover and in antioxidant defenses. Therefore, SIRT3 could confer resistance to chemotherapy-induced oxidative stress, leading to a lower ROS production and a higher cell survival. Our aim was to analyze whether SIRT3 silencing in breast cancer cells through a specific siRNA could increase oxidative stress and thus compromise the antioxidant response, resulting in a sensitization of the cells to cisplatin (CDDP) or tamoxifen (TAM). For this purpose, we studied cell viability, ROS production, apoptosis and autophagy in MCF-7 and T47D cell lines treated with these cytotoxic compounds, these either alone, or in combination with SIRT3 silencing. Moreover, protein levels regulated by SIRT3 were also examined and survival curves were analyzed to study the importance of SIRT3 expression for the overall survival of breast cancer patients. When SIRT3 was silenced and combined with cytotoxic treatments, cell viability was highly decreased, and was accompanied by a significant increase in ROS production. While TAM treatment increased autophagic cell death, CDDP significantly triggered apoptosis, whereas SIRT3 silencing produced an enhancement of these two action mechanisms. SIRT3 knockdown also affected PGC-1α and TFAM (mitochondrial biogenesis), and MnSOD and IDH2 (antioxidant defenses) protein levels. Finally, survival curves showed that higher SIRT3 expression is correlated to a poorer prognosis for patients with grade 3 breast cancer. In conclusion, SIRT3 could be a therapeutic target for breast cancer, improving the effectiveness of CDDP and TAM treatments.