Proteomic Analysis to Identify Functional Molecules in Drug Resistance Caused by E-Cadherin Knockdown in 3D-Cultured Colorectal Cancer Models.

摘要

In the second year of research, we successfully optimized the SILAC methods and phosphoproteomic analysis methods. The optimized method has increased the phosphopeptide identification for 2 folds. We also optimized the system for testing drug effects. Four different drugs including irinotecan, oxaliplatin, 5-FU, and gefitinib were tested with CDH1 knock down cells for drug resistance effects. Cells with down regulated CDH1 levels show resistance to all drugs at high concentration except 5-FU. The mass spectrum data for drug resistance pathways are under collection and investigation. We also analyzed the proteomic and phosphoproteomic differences between 2D and 3D cultured cells. In conclusion, during the second year, we accomplished most of the work we proposed in our original proposal, and finished the tasks that got delayed during the first year. The second year s research is a good extension of the first year research, and provided us the foundation for our future work.