P04.05 The Effects and Mechanism Involved in Paclitaxel combined with Shikonin on U87 Glioma Cells

Abstract BACKGROUND Glioma is the most common intracranial primary tumor,chemotherapy seems to be the main treatment for brain glioma.Chemotherapy is currently widely used in clinical treatment of gliomas,however,drug-resistant always appears,so combination of chemotherapy has became an inevitable choice. Paclitaxel and shikonin are stable and efficient chemotherapy drugs which can promote apoptosis and ininhibite the growth of tumor.Recent research reports,paclitaxel and shikonin can respectively activate and inhibit the expression of p-ERK.The purpose of this study was to investigate whether we can inhibite the activation of p-ERK caused by paclitaxel to enhance the effect of paclitaxel,furthermore use the combination of the two drugs to reach a better effect of chemotherapy. MATERIAL AND METHODS 1.Cell Culture2. Drugs Administration.3. Growth Inhibition Assays (CCK-8) .4.Scratch test to observe cell migration 5.Transwell assay for the determination of cell migration 6.Transwell assay for the determination of cell invasion 7. Annexin V-FITC and PI Assay for Flow Cytometry to Detect Apoptosis 8. PI Assay for Flow Cytometry to Detect Cell Cycle 9. Western Blot to Detect the Expression of P-ERK/ERK 10. Statistical Analysis. RESULTS 1.The results tested by CCK-8 showed these two drugs have synergic effects with a CDI equals to 0.72 which means these two drugs are significant synergistic.2.Compared with paclitaxel group or shikonin group,the 1/2paclitaxel+1/2shikonin group had a significant inhibitory effect upon the migration of U87 cells.3.Compared with paclitaxel group and shikonin group,the 1/2paclitaxel+1/2shikonin group had a significant inhibitory effect upon the invation of U87 cells.4.Compared with paclitaxel group and shikonin group,the 1/2paclitaxel+1/2shikonin group had a significant promoting effect upon inducing apoptosis of U87 cells.5. The 1/2paclitaxel+1/2shikonin group significantly improved the apoptosis rate of U87 cells,the RNA content of the G1-phase was significantly increased in shikonin group, the RNA content of the G2- phase significantly increased in paclitaxel group and 1/2paclitaxel+1/2shikonin group.6.Compared with control group,the rate of p-ERK/ERK was increased in paclitaxel group(P<0.05),while the rate of p-ERK/ERK was significantly decreased in the shikonin group and 1/2paclitaxel+1/2shikonin group(P<0.05). CONCLUSION 1. Compared with paclitaxel group and shikonin group,the combination group with half dosage can make synergic effects and significantly inhibit the proliferation,migration and invation ability of U87 cells,besides promote apoptosis of U87 cells and block U87 cell cycle.2. Paclitaxel combined with shikonin could decrease the expression of p-ERK/ERK,which may be involved in the synergic effects of these two drugs.