Study On The Molecular Mechanisms Of Osteosarcoma Cell Apoptosis Induced By Adriamycin In Combination With Microwave

Surgical operation still serves as our current major method for treating bone tumor, aided with chemotherapy, radiotherapy, interventional therapy and immunotherapy. Adjuvant chemotherapy can improve five-year survival rate and remaining limbs rate among bone tumor patients, but in most cases, simplification chemotherapy for treating bone tumor is not enough. Thermotherapy, especially microwave thermotherapy, is testified to have a significant effect to improve remaining limbs rate. Vast previous reports have shown that the combination of chemo-thermotherapy is mutual complementation and synergistic effect, and will have a great prospect in future tumor treatment.Clinical application of operation-ADM-microwave combine therapy has proved to be effective and successful in treating osteogenic sarcoma, the most fatal and common bone tumor, but detail mechanism of action of thistreatment remains unclear. In this research, we analyzed the apoptosis effect of AMD, microwave and AMD-microwave combination to human osteogenic sarcoma cells through DNA laddering, flow cytometry and soft agar colony forming experiment. We found that time and concentration relied cell apoptosis occurred after all the three treatment, and the combine use of ADM and microwave is most effective in inducing cell apoptosis, and AMD is after microwave.We used several signaling molecule inhibitors to test which special signal transduction pathway or signaling molecules worked during cell apoptosis in these three groups. Except for P-38 in inhibitor SB203580 and JNKK 1/3 inhibitor PD98059 that could improve cell reproductive activity and even recover to normal level, no other inhibitors proved effective. It means these two signal transduction pathways should be connected with the apoptosis of osteogenic sarcoma.We used both phosphorylation and no phosphorylation specific antibodies aiming directly at MAPK family members of JNK, ERK, p38 to detect the changes of express and activation level about these three treatment, through western blot. We found there were no apparente changes of these kinases express level, but apparente changes of these kinases phosphorylation degree in sharp contrast to control group. The phosphorylation degree of ENK was inhibited, and the phosphorylation degree of JNK, p38 was increased. Because kinase phosphorylation degree embodies its activation level, we got a conclusion that three treatment suppressed the activity of ENK and increased the activity of JNK, p38. We also presumed that this cellular survival condition or apoptosis degree would have determinate associativity with accommodative effect of MAPK family members of JNK , p3 8.