Preliminary Study And Safety Evaluation Of The Inhibition Of Nasopharyngeal Carcinoma Cell Line CNE-1 By Total Flavonoids From Rhizoma Aristolochiae

Objective:In this paper,in vivo and in vitro model experiments were conducted to investigate the effects of total flavonoids of Lycium barbarum L.on the proliferation,cloning,scratching,migration,invasion,apoptosis,cycle and anti-tumor effects of nasopharyngeal carcinoma cell line CNE-1;Western blot was used to detect the expression of related proteins,and then the mechanism of total flavonoids from Rhizoma Aristolochiae on nasopharyngeal carcinoma cell line CNE-1 was elucidate preliminarily.In order to evaluate its safety,the long-term and acute toxicity of total flavonoids from Rhizoma Lobeliae were observed by intragastric administration in mice and rats,which provided a theoretical basis for improving the utilization value of total flavonoids in Rhizoma Lobeliae and developing new medicines.method:1.MTT assay,single cell cloning assay,cell scratch assay,cell invasion assay,cell cycle apoptosis assay were used to detect the effects of total flavonoids?RDB?in vitro on the proliferation,cloning,migration,invasion,apoptosis and cycle of CNE-1 cells,which led to arrive at the concentration of RDB in Rhizoma Dioscoreae had an effect on CNE-1 cells.2.The therapeutic effect of RDB on nasopharyngeal carcinoma was observed by in vivo model of nasopharyngeal carcinoma nude mice.3.whether RDB alter the expression of related proteins was observed by western blot.4.The safety of RDB was evaluated by acute toxicity test and long-term toxicity test of RDB.result:1.The IC₅₀0 of CNE-1 cells treated with RDB for 24h,48h and72h was 63.23?g/ml,54.50?g/ml and 41.20?g/ml,respectively.The concentration of RDB was 30?g/ml by single cell cloning assay.The cell clone survival rate was 49.25%and 22.5%at 60?g/ml,respectively.The healing rate of CNE-1 cells treated with 30?g/ml and 60?g/ml at 48 h was 44.01±3.54%,respectively.And 37.96±2.32%;CNE-1 cells with a RDB concentration of 30?g/ml through the chamber at 72 h were only28.3%of the control group,and the concentration of RDB was 60?g/ml through the chamber.-1 cells only contained 1.1%of the control group;the apoptosis rate of the CNE-1 cells reached 55.58%after 72 hours of RDB concentration of 60?g/ml.Through the measurement of cell cycle,the apoptosis rate of CNE-1 cells treated with RDB increased significantly,and cell block occurred in G0/G1 phase.2.In vivo experiments in nude mice showed that RDB had significant inhibitory effect on nasopharyngeal carcinoma cell line CNE-1in nude mice.The inhibition rates of high,medium and low doses of RDB after 21 days of administration were 52.15%+14.65%,40.61%+17.51%,37.89%+12.79%,respectively.3.Western blot analysis showed that the expression of p53,p21 and caspase-3 was up-regulated and the expression of Bcl-xL and Bcl-2 was down-regulated by RDB in nasopharyngeal carcinoma cell line CNE-1.4.According to the acute toxicity test and long-term toxicity test of RDB,no mice died during the acute toxicity test,and no obvious pathological changes were found in the liver sections of mice after administration.During the long-term toxicity experiment,rats took RDB for a long time,but their body weight,organ index,pathological section and serum index were normal except sodium ion.conclusion:1.RDB could inhibit the proliferation and cloning of nasopharyngeal carcinoma cell line CNE-1,and reduce its migration and invasion ability,block the cell cycle at G0/G1,and promote the apoptosis.2.RDB could effectively inhibit the growth of nasopharyngeal carcinoma CNE-1 cells in nude mice.3.CNE-1 cells treated with RDB could induce apoptosis of nasopharyngeal carcinoma CNE-1 cells through Bcl-2 family,caspase family and p53 signaling pathway.4.The acute and long-term toxicity studies of RDB indicate that RDB was safe to take orally.