Levobupivacaine Inhibits Proliferation And Induce Apoptosis In Breast Cancer Cells Through The PI3K/AKT/mTOR Signaling Pathway.

Purpose:This study aimed to investigate the effect of levobupivacaine on breast cancer cell proliferation and apoptosis.Background:Breast cancer is the most prevalent cancer and is responsible for many deaths in women.In this regard the urgent need to find new therapeutic agents to improve breast cancer treatment is imminent.Recent studies have demonstrated that local anesthetics exhibit an anti-tumor effect in cancer cells.This study aimed to investigate the inhibition effect of levobupivacaine on breast cancer cells thus promoting apoptosis through molecular signaling pathway.Methods:Human breast cancer cells,MCF-7 and MDA-MB 231 were cultured and treated with 1mM and 2mM concentrations of levobupivacaine for 24hr.Cell survival was assessed by cck-8 assay.The proliferation was determined by western blot,qPCR,colony formation and transwell assay.Apoptosis was measured by annexin V and PI staining on flow Cytometry.The effects of levobupivacaine on cellular signaling and molecular response were studied with quantitative PCR assay and western blot.Results:Induction of Apoptosis was confirmed by cell viability,morphological changes showed cell shrinkage,rounding,and detachments from plates.Results of western blot and qPCR downstream indicated activation of Caspase-3 and inhibition of Fox01.The results of the flow cytometry confirmed that levobupivacaine induced cell cycle arrest in a dose-dependent manner which resulted an increase in apoptosis.Also,the activation of p21 and a decrease in cyclin D expression in qPCR and Western blot confirmed the inhibition of cells’ ability to proliferate compared with the control.However,the qPCR and western blot also showed that Levobupivacaine could significantly up-regulate the expression of Bax,accompanied by a significant down-regulation of Bcl2 expression and decrease in protein expressions of PI3K-AKT-mTOR upstream when phosphorylated.Conclusions:These findings suggested that Levobupivacaine at significant concentrations inhibits proliferation and promotes apoptosis in breast cancer in vitro.