The Influence Of Tmz On The Expression Of Livin,mrp1 And Mrp3 In Plant Tumor Of Glioma Stem Cells

ObjectGlioma is a kind of common and dangerous intracranial malignant tumor. Even accepted combination treatment including surgery, radiotherapy and chemotherapy, the middle survival time of glioblastoma patients were only 12 to 18 months. We can not cure even the low grade glioma. One of the most important reason lies in the chemotherapy drugs resistance of glioma. Therefore, further study of etiology, pathogenesis and chemotherapy drug resistance has important clinical and social value. In recent years, Cancer stem cells were thought to be the basis of tumor occurrence, development and maintain, and were related to tumor recurrenc, metastasis and resistance to drug. Research showed that chemotherapy drug resistance of glioma stem cells was stronger than glioma cells. The main reason may because glioma stem cells could generate strong chemotherapy drug resistance and eventually led to the tumor recurrence. Therefore, researching glioma chemotherapy drug resistance by cancer stem cells model is likely to further clarify the mechanism of tumor chemotherapy drug resistance and produce new medicine for glioma.ContentThis topic was focused on the expression rule of Livin, MRPl and MRP3 in tumors of U251 cells and it’s stem cells. Analysis possible mechanism of glioma stem cells chemotherapy drug resistance.We cultivated U251 cells and separated CD133 + stem cells. Making tumor models in nude mice brain. Researching the effect of TMZ on the expression of Livin, MRP1 and MRP3.Method1. CD133 positive stem cells were isolated from U251 cells line by immune magnetic bead. 2. Tumor models in nude mice brain of U251 cells and it’s stem cells were built by brain stereotactic technology and Trace injection technology. Mice were kepted for 12 days, then intervented with TMZ for six days. The change of weight, growth and behavior of mice were observed. 3.Mice were executed, tumor samples were collected and tumor pathology were observed by paraffin embedding, HE staining and biopsy and immunohistochemical staining. 4.RT-PCR was used to detect the expression of livin, MRP1 and MRP3 in U251 cells and stem cells tumors.Result1. U251 stem cells presented suspended, spherical growth in serum-free medium withthe typical "stem cells" characteristics. U251 stem cells expressed positive after Nestin testing. After differentiation in medium containing serum, the cells presented positive after GFAP and β-tubulin testings. 2. The weight of mice in U251 stem cells group loss sooner and faster than U251 cells group and cachexia appeared stronger. 3.There was no obviously boundaries between U251 stem cells tumor and brain tissue. There was obviously boundaries between U251 cells tumor and brain tissue. The expression of GFAP in U251 stem cells tumor was stronger than U251 cells tumor. 4.RT-PCR showed the expression of Livin m RNA in U251 stem cells tumor was obviously higher than U251 cells tumor(p<0.05). The expression of MRP1 m RNA in U251 stem cells tumor was obviously higher than U251 cells tumor(p<0.01). The expression of MRP3 m RNA in U251 stem cells tumor was obviously lower than U251 cells tumor(p<0.01).With the increase of TMZ concentration, the expression of MRP1 and MRP3 m RNA in U251 stem cells tumor enhanced(p<0.01). The expression of MRP1 m RNA in U251 cells tumor enhanced While MRP3 m RNA reduced(p<0.01). The difference was statistically significant.Conclusions1.Weight of mice in U251 stem cells group loss sooner and faster than U251 cells group, Cachexia and limb hemiplegia appeared. It showed malignance of U251 stem cells tumor was stronger. 2.There was no obviously boundaries between U251 stem cells tumor and brain tissue. There was obviously boundaries between U251 cells tumor and brain tissue.It showed invasiveness of U251 stem cells tumor was stronger than U251 cells tumor. 3.Multiple drug resistance gene MRP1, MRP3 played different roles in chemotherapy resistance mechanism of U251 cells tumor and U251 stem cells tumor. MRP1 may play a more important role in U251 cells tumor while MRP1 and MRP3 may play synergy in U251 stem cells tumor.