Study On The Cytotoxic Effect And Mechanism Of Ampelopsin Sodium Combined With Carboplatin On Human Lung Adenocarcinoma Cell Line SPC-A-1 In Vitro And Vivo

Objective:The present study aimed at investigating the cytotoxic effect and mechanism of ampelopsin sodium(AMP-Na) on human lung adenocarcionma cell line SPC-A-1 by alone or combined with carboplatin in vitro and vivo.Methods:SPC-A-1 cells were injected subcutaneously at right back of nude mice to establish transplanted human lung cancer model and to evaluate the anti-tumor effect of ampelopsin sodium alone or combined with carboplatin in vivo.The inhibitory effects on proliferation of human lung cancer cell line SPC-A-1 were examined by MTT colorimetric assay.Flow cytometry(FCM) was employed to analyze the cell apoptosis,and to detect the expression of Bcl-2,Bax and Caspase-3.Results:1.The experiment in vivo showed that ampelopsin sodium 125～260 mg/kg used alone has not a manifest anti-tumor effect on the transplanted human lung cancer cell line SPC-A-1 in nude mice.But when ampelopsin sodium combined with carboplatin, the anti-tumor activity of carboplatin was significantly increased,especially 200 mg/kgⅠand 260 mg/kg as compared with that in control group and in group treated with carboplatin alone,suggesting there was a synergistic anti-tumor action of AMP-Na and CBP.2.Experimental study on cytotoxicity in vitro showed that AMP-Na decre ased the survival rate of SPC-A-1 cells in a concentration-dependent manner ra nging from 6.25μg/ml to 200μg/ml.There was a synergistic cytotoxic effect on SPC-A-1 cells treated with AMP-Na combined with carboplatin.3.FCM analysis showed that ampelopsin sodium 12.5～100μg/ml significantly induced apoptosis of SPC-A-1 cells in vitro.The apoptosis rate in group treated with AMP-Na combined with carboplatin was significantly increased as compared with that in group treated with carboplatin alone.4.FCM analysis showed that the fluorescence intensity of Bcl-2 and Bax all increased after SPC-A-1 cells treated with AMP-Na alone or combined with carboplatin,the ratio of Bcl-2/Bax was decreased,and the positive express ratio of Caspase-3 was significantly increased.Conclusion:1.The result of experiment in vivo shows that ampelopsin sodium used alone has not a manifest anti-tumor effect,while combined with carboplatin,it can increase the anti-tumor effect of carboplatin on the transplanted human lung cancer of SPC-A-1 cell line in nude mice.2.AMP-Na decreases SPC-A-1 cells survival rate in a concentration-dependent manner,and shows significant cytotoxic activity.There is a synergistic cytotoxic effect on SPC-A-1 cells treated with AMP-Na combined with carboplatin.3.The mechanism of AMP-Na to induce SPC-A-1 cells apoptosis is probably that it can activate Caspase-3 mediated signal transduction pathway in cells and decrease the ratio of Bcl-2/Bax.