[Effects of ropivacaine hydrochloride on the proliferation, invasion and apoptosis of osteosarcoma cells and its mechanism].

ABSTRACT

Objective: To investigate the effects and molecular mechanism of ropivacaine hydrochloride on osteosarcoma cell proliferation, invasion, and apoptosis. Methods: The osteosarcoma doxorubicin-resistant cell line (U2OS/DOX) was established by gradually increasing the drug doses. U2OS/DOX cells were treated with ropivacaine hydrochloride at the concentrations of 0, 20, 50 and 100 µg/ml, respectively; as different concentrations treatment groups of ropivacaine hydrochloride. pcDNA3.1 and pcDNA3.1-Livin were transfected into U2OS/DOX cells and then treated with 100 µg/ml ropivacaine hydrochloride, which were defined as ropivacaine hydrochloride 100 µg/ml+pcDNA3.1 group, ropivacaine hydrochloride 100 µg/ml+pcDNA3.1-Livin group. MTT was used to detect the cell proliferation inhibition rate and inhibitory concentration (IC50). Western blot was used to detect the expressions of cyclin-dependent kinase inhibitor 1A (P21) and activated cysteine aspartic protease-3 (Cleaved Caspase-3), E-cadherin, matrix metalloproteinase 2 (MMP-2) and Livin; clone formation experiments were used to detect the number of cell clones formed; flow cytometry was used to detect apoptosis; Transwell was used to detect cell migration and invasion; real-time fluorescent quantitative PCR (RT-qPCR) was used to detect Livin mRNA expression. Results: When the concentration of doxorubicin was more than 1 µg/ml, the proliferation inhibition rate of osteosarcoma cells U2OS was significantly increased in a concentration-dependent manner (P＜0.05); when the concentration of doxorubicin was more than 10 µg/ml, the proliferation inhibition rate of osteosarcoma resistant cell U2OS/DOX was significantly increased, and it was dose-dependent (P＜0.05). In U2OS/DOX cells treated with ropivacaine hydrochloride, the expressions of P21, Cleaved Caspase-3, and E-cadherin were increased significantly, the expression of MMP-2 was decreased significantly, the cell proliferation inhibition rate was increased significantly, the number of colony formation was decreased significantly, and the cells apoptosis rate was increased significantly, the number of cell migration and invasion was decreased significantly, and the expression of Livin was significantly reduced, in a concentration-dependent manner (P＜0.05). Overexpression of Livin partially reversed the inhibitory effect of ropivacaine hydrochloride on proliferation, migration, invasion, and promotion effect on apoptosis of cell U2OS/DOX. Conclusion: Ropivacaine hydrochloride can significantly inhibit the proliferation, migration and invasion of doxorubicin-resistant osteosarcoma cells, and significantly promote osteoma cell apoptosis. The mechanism may be related to Livin.