The Empirical Study Of Chemotherapy With Nimustine Plus Cisplatin For C6 Glioma

Object.The brain glioma is the most common central nervous system primary tumor; the treatment of glioma is a difficult problem that has been perplexing in the field of department of neurosurgery for a long time.Especially advanced malignant glioma growed widespreadly,invading,with unclear border,high recurrence of postoperative, devastating prognosis and high mortality.The biological characteristics of glioma result in complex treatment being adopted in treatment,which means surgery, cooperated with radiotherapy,chemotherapy,immunotherapy,etc.while being postoperative.The position in the tumour treatment of chemotherapy has been already clear and definite.But we often find chemical therapeutic was unsatisfactory in clinical work,because of the drug resistence gene MGMT and blood brain barrier existence,which serious influence chemotherapy function normally.We aimed to explore the effects of ACNU combined with CDDP in glioma treatment by experiment in vitro and rat glioma xenograft,and analyze clinical feasibility and validity of drug combination,compared with the curative effect of single drug ACNU or CDDEMethod:C6 glioma cells were cultured into 4 groups at random.Control group, ACNU group,CDDP group,and ACNU+CDDP group.We get the amount of drug used in the experiment through the formula referring to dosage of the clinical medicine;four groups were offered chemotherapy medicine separately.Cell cycle and cell apoptosis was analyzed by flow cytometry(FCM) skill and dyed with Annexin V-FITC/PI.The establishment of Wistar rats with intracranial C6 glioma model were confirmed successful by Gd-DTPA enhanced MRI scan,and then rats were randomly divided into 4 groups as same as above.After delivery for 3 days, some rats of each group were sacrificed,HE staining was used to observed morphology of tumor tissue,immunohistochemistry examined level of a proliferation-related protein PCNA and apoptosis- -related protein bcl-2.By the way, the survival of the remaining rats' situation was observed.Results:Cells in ACNU+CDDP group were more sensitive to the medicine than other three group,we can see that cells died in a large amount,the surplus cell shape is obviously thin and weak.FCM analysis showed no obvious change in cell cycle of each group,but cell apoptosis rate of ACNU+CDDP group was 2.1%,which increased a lot compared with others(p＜0.01).Rats with C6 glioma showed less activity and diet,dispirited spirit,late dysfunction gradually.Its survival time was 29.41 days on average,the survival analysis reveals life cycle of group ACNU+CDDP was obviously longer than three other groups'.Pathology detection showed significant patterns of cell shapes,large nuclear with deeply stained,and cell polar permutation,irregular,also the existence of large areas of apoptosis and necrosis in ACNU + CDDP group.Discrete tumor foci counting analysis of the margin revealed ACNU + CDDP group is much less than the remaining three groups, with significant statistically differences.Immunohistochemical test showed PCNA-positive located in the nuclei,expression of ACNU + CDDP group was the lowest,about 44.84%;Bcl-2 protein located in the cytoplasm,the expression of ACNU + CDDP group was also the lowest(p＜0.01).Conclusions:In this study,both cell and animal experiment showed ACNU + CDDP combination is more than single drug group inhibiting glioma cell proliferation and promoting apoptosis,which indicated that the two drug combination therapy was superior to the effect of glioma using single ACNU or CDDP.In clinical studies we also found that ACNU combined with CDDP delivered injectively for 72 hours can effectively inhibit tumor growth,but its severe bone marrow suppression influence the repeated chemotherapy.Histopathology in this study also revealed that there were a large number of tumor necrosis in the combination group,and more importantly, the number of discrete tumor foci around could be significantly reduced.According to the above conclusions,we consider the possibility of ACNU+CDDP treatment as adjuvant chemotherapy prior to surgical,so as to reduce tumor recurrence rates, extended survival time,so that patients with no surgery opportunities because of focus locating in the hypothalamus,brain stem,motor areas and other important functional areas could get chance to take operation.Neoadjuvant chemotherapy may bring new hope for glioma treatment.