Study On Anti-tumor Effects And Its Mechansim Of Ampelopsin Sodium Combined With Carboplatin On The Human Lung Adenocarcinoma GLC-82 Cells

Objective:1.To investigate the inhibitory effect of Amp-Na alone and combined with carboplatin on the proliferation of human lung adenocarcinoma GLC-82 cells. To study the effective dose of ampelopsin sodium (Amp-Na) alone and combined with carboplatin.2.To study the mechanism of the anti-tumor effect of Amp-Na.3.To investigate the therapeutical effect of Amp-Na and Amp-Na combined with carboplatin on growth of transplanted lung adenocarcinoma GLC-82 in nude mouse.Methods:1.Use The MTT reduction assay(Mono-nuclear cell direc cytotoxicity assay) to investigate cytotoxicity effection of Amp-Na alone and combined with carboplatin.2.Use flow cytometry (FCM) to detect apoptosis of lung adenocarcinoma cell GLC-82,and also detect expression of caspase-3,Bax and Bcl-2.3.To study the therapeutical effect of Amp-Na alone or combined with carboplatin on growth of transplanted lung carcinoma GLC-82 in nude mouse in vivo.Results:1.Proliferation of human lung adenocarcinoma GLC-82 cells was inhibited by Amp-Na alone in a concentration-dependent manner ranging from 12.5-200μg/ml,and it's IC50 was 56.16μg/ml.2. Proliferation of human lung adenocarcinoma GLC-82 cells was inhibited by carboplatin alone in a concentration-dependent manner ranging from 3.13-100μg/ml,and it's IC50 was 39.93μg/ml.3. Combined with carboplatin, Amp-Na 50 and 100μg/ml had synergistic effect on the proliferation of GLC-82 cells .4.Flow cytometry analysis on GLC-82 cell showed that either treated by Amp-Na alone or Combined with carboplatin, apoptotic cells and necrotic cells all increased The expression of caspase-3 and Bax were increased significantly after treated with Amp-Na or combined with carboplatin for 48 hours;The ratio of Bcl-2 and Bax was decreased significantly after treated with Amp-Na alone or combined with carboplatin for 48 hours.5.The experiment indicate that there was no effect on human transplanted lung adenocarcinoma GLC-82 in nude mouse treated with Amp-Na alone and treated with carboplatin alone ; When combined with carboplatin, Amp-Na 50,80,125,200 mg/kg had synergistic depress effect on growth of transplanted lung adenocarcinoma GLC-82 in nude mouse in vivo.Conclusion:1.The inhibitory effect on the proliferation of GLC-82 is observed in groups treated with Amp-Na and carboplatin in a concentration-dependent manner.2. Combined with carboplatin,Amp-Na has synergistic effect on the proliferation of GLC-82 cell.3.Apoptotic cells and necrotic cells are found on GLC-82 cell treated with Amp-Na alone or combined with carboplatin.One of the mechanisms to induce apoptosis by Amp-Na is probably that activation of caspase-3 mediated signal transduction pathway in cells.4. Amp-Na and carboplatin alone have no effect on growth of transplanted lung carcinoma GLC-82 in nude mouse, and when Amp-Na combined with carboplatin there is synergistic depress effect on the growth of transplanted lung adenocarcinoma GLC-82 in nude mouse.