Colony formation and transwell assay were used to determine
breast cancer cells proliferation. Flow Cytometry (
annexin V and PI staining) was used to investigate
breast cancer cells apoptosis. The effects of
levobupivacaine on cellular signalling and molecular response were studied with Quantitative Polymerase Chain Reaction and western blot. Induction of apoptosis was confirmed by cell viability, morphological changes showed cell shrinkage, rounding, and detachments from plates. The results of the western blot and Quantitative Polymerase Chain Reaction indicated activation of active
caspase-3 and inhibition of FOXO1. The results of the flow Cytometry confirmed that
levobupivacaine inhibited
breast cancer cell proliferation and enhanced apoptosis of
breast cancer cells. Quantitative Polymerase Chain Reaction and Western blot analysis showed increased p21 and decreased
cyclin D. Quantitative Polymerase Chain Reaction and western blot analysis showed that
levobupivacaine significantly increased Bax expression, accompanied by a significant decreased Bcl-2 expression and inhibition of PI3K/Akt/mTOR signalling pathway. These findings suggested that
levobupivacaine inhibits proliferation and promotes
breast cancer cells apoptosis in vitro.