Differential Analysis Of Two-dimensional Gel Electrophoresis Profiles Of Proteins In Multidrug Resistant Human Hepatocellular Carcinoma Cell Line HepG2/ADM And Its Counterpart HepG2

Objective Primary hepatocellular carcinoma (HCC) which has high malignancy level, high mortality and poor prognosis, is one of the most common malignant tumors in China and recently the morbidity of HCC tends to go rising. The Individual combined therapy which mainly consisted of surgery has been became the standard treatment plan for HCC at present. Chemotherapy, as a critical adjuvant treatment method, still play an important role in clinical therapy of HCC. Although many novel chemotherapeutic drugs and plans emerged constantly, the effect of chemotherapy for HCC has been not improved conspicuously. The main reason is multidrug resistance (MDR), which means once the cancer cells are resistant to one chemotherapeutic drug and they are also cross resistant to the other chemotherapeutic drugs which have different structures and different mechanisms of action from the former one. Therefore the MDR is an urgent problem need to be solved for malignant tumors treatment. Our objective in this research is to establish two-dimensional gel electrophoresis profiles of proteins in multidrug resistant human hepatocellular carcinoma cell line HepG2/ADM and its counterpart HepG2, and to analyze the differences in proteins expression between these two cell lines.Methods MTT assay was used to determine the sensitivity of HepG2/ADM and HepG2 to different chemotherapeutic drugs. The total proteins of HepG2/ADM and HepG2 were separated by two-dimensional gel electrophoresis and visualized by silver staining. The differences in proteins expression between these two cell lines were analyzed by ImageMaster 2D Platinum software.Results The HepG2/ADM was demonstrated conspicuous multidrug resistant compared with HepG2. The basic fame distributions of total proteins in the two cell lines were similar. 45 differential protein spots expressed in these two cell lines were distinguished, 25 protein spots were up-regulated in HepG2/ADM,9 protein spots were down-regulated in HepG2/ADM, 7 protein spots expressed only in HepG2/ADM, 4 protein spots expressed only in HepG2.Conclusion The two-dimensional gel electrophoresis profiles with high resolution and reproducibility of total proteins in HepG2/ADM and HepG2 were established, and some differential protein spots were found. The data obtained from this study prepared the ground for the further study on multidrug resistance of hepatocellular carcinoma by proteomic methods.