The Effects Of AZT On The Telomerase Activity And Proliferation In Human Brain Glioma Stem Cell

Objective:To investigate the effects of AZT on the human brain glioma stem cells and malignant glioma cells, so as to gain further insight about the therapy of malignant glioma.Methods:1. Isolated from fresh specimens of brain and cultured brain glioma stem cells and glioma cells.2. We use Cellular immune fluorescence to detect tumor stem cells ball of tumor-specific markers CD 133 and nestin.3. The experiment was divided into experimental group (0.5mol/L,0.25mol/L 0.125mol/L) and control group.We performed immunocytochemistry of BTSCs for CD 133 and nestin expression.MTT method was used to detect the cells survival rate. Flow cytometry(FCM) was used to examine the cell apoptosis rate and phases of cell cycle. Telomerase activity was determined by TRAP-PCR-ELISA.Results:1. A few cancer cells can survive in serum-free media and suspension proliferate, and form cloned glioma stem cell ball. After passage, brain glioma stem cells remain strong self-renewal and proliferative capacity, express the tumor-specific markers CD 133 and nestin.2. AZT function 24h,0.25mol/L group of glioma stem cell proliferation was inhibited (P<0.05),0.125mol/L group of glioma cell proliferation was inhibited (P<0.05), 0.5mol/L group brain tumor stem cells appear significantly different (P<0.05). 3. AZT function 24h, the 0.5mol/L group telomerase activity in glioma stem cells decreased (P<0.05); After 24h, the 0.25mol/L group telomerase activity in brain glioma cells decreased (P<0.05). In the experimental range, it is showing the time-dose dependent manner.4. AZT function 72h, the apoptosis of glioma stem cell peak was 4.21% (0.25mol/L),10.6%(0.50mol/L), peak of apoptosis of glioma cell was 6.75% (0.25mol/L),14.3%(0.50mol/L).Conclusion:1. This experiment preliminary confirmed that the brain glioblastoma multiforme have a brain tumor stem cells existed in, we can get it by the serum-free culture method.2. The markers neural stem cells of nestin and CD 133 can be used to identify brain stem cells.3.AZT can inhibit the activity of brain glioma stem cell and glioma cell.Compared with glioma cell, brain glioma stem cell was resistant to AZT.