The Influence Of Temozolomide On Livin And ABCG2 In Human Brain Glioma TJ905 And Stem Cells

Background:Our study is designed to detect anti-apoptosis Livin gene expression in TJ905 cells of human brain glioma, to observe the anti-tumor effects in vitro of chemotherapy drugs temozolomide on human brain glioma TJ905 cells and glioma stem cells, and to study the extent of glioma’s drug resistance by gioma cells and glioma stem cells as a modeol. So far, the quantity of research reports on the relationship between anti-apoptosis of glioma cells and giloma stem cells induced by anti-apoptosis Livin gene and drug resistance and its mechanism is still small.Objective:We are trying to reveal the roots of glioma drug resistance, enrich and develop the theoretical basis of cancer cells and stem cells by analysing human brain glioma TJ905 cells and glioma stem cells which control by temozolomide, detecting the changes in cell survival、cell cycle and the expression levels of Livin mRNA,drug resistance gene ABCG2 mRNA after intervention. And then, providing new ideas and methods for the treatment of glioma and laying a solid theoretical and experimental basis to overcome chemotherapy resistance in the treatment of glioma difficulties.Methods:The CD 133+positive TJ905 stem cells from human brain glioma TJ905 cells were isolated by using immune magnetic beads and identified by CD 133 antibody. To found the glioma cells and stem cells models. The changes in cell migration, the trend of cell proliferation, the changes in cell cycle and the expression of Livin mRNA, ABCG2 mRNA were detetced by using cell scratch technique CCK-8、flow cytometry and RT-PCR after the intervention with different drug concentrations(050,100,200,400 μmol/1) at different time points (24 h,48 h and 72 h).Results:1. At the same intervention time, the migration of human brain glioma TJ905 cells decreased with increasing of drug concentration (P<0.05). At the same temozolomide concentration, the migration of human brain glioma TJ905 cells decreased with increasing of intervention time (P<0.05).2. With increasing temozolomide concentration and intervention time, the density of human brain glioma TJ905 cells decreased, human brain glioma TJ905 cells shrinked, the proliferation of human brain glioma TJ905 cells was inhibited, scattered debris of human brain glioma TJ905 cells appeared and stem cells cracked and contracted.3. At the same intervention time, the survival rates of human brain glioma TJ905 cells and stem cells decreased with increasing of temozolomide concentration (P<0.05). At the same temozolomide concentration, the survival rates of human brain glioma TJ905 cells and stem cells decreased with increasing of intervention time (P<0.05). The survival rates of human brain glioma TJ905 cellss were lower than those of the stem cells.4. The cell cycle of human brain glioma TJ905 cells arrested in the GO-G1 period and G2-M period with temozolomide intervention. At the same intervention time, the proportion of human brain glioma TJ905 cells that arrested in the GO-G1 period and G2-M period was bigger and bigger with increasing of temozolomide concentration (P<0.05). At the same temozolomide concentration, the proportion of human brain glioma TJ905 cells that arrested in the GO-G1 period and G2-M period was bigger and bigger with increasing of intervention time (P<0.05). The proportion of human brain glioma TJ905 cells that arrested in the GO-G1 period was higher than in the G2-M period.5. The cell cycle of TJ905 stem cells arrested in the G2-M period and S period with temozolomide intervention. At the same intervention time, the proportion of human brain glioma TJ905 stem cells that arrested in the G2-M period and S period was higher and bigger with increasing of temozolomide concentration (P<0.05). At the same TMZ concentration, the proportion of human brain human brain glioma TJ905 stem cells that arrested in the G2-M period and S period was bigger and bigger with increasing of intervention time (P<0.05). The proportion of human brain glioma TJ905 stem cells that arrested in the S period was higher than in the G2-M period.6. At the same intervention time, Livin mRNA expression in human brain glioma TJ905 cells and stem cells was lower and lower while ABCG2 mRNA expression higher and higher with increasing of TMZ concentration (P<0.05). At the same temozolomide concentration, Livin mRNA expression in human brain glioma TJ905 cells and stem cells was lower and lower while ABCG2 mRNA expression higher and higher with increasing of intervention time (P<0.05). With increasing temozolomide concentration and intervention time, the Livin mRNA expression in human brain glioma TJ905 cells was higher than that in TJ905 stem cells, but the and ABCG2 mRNA expression was higher than that in TJ905 cells; The Livin mRNA rate of falling in human brain glioma TJ905 cells was higher than that in TJ905 stem cells, The ABCG2 mRNA rate of rising in human brain glioma TJ905 cells was lower than that in TJ905 stem cells..Conclusion:The expression of Livin mRNA, glioma cells and stem cells cycle arrest, proliferation speed were inhibited after intervention with temozolomide. However, the expression of ABCG2 mRNA, glioma cells and stem cells apoptosis enhanced. Human brain glioma TJ905 stem cells have stronger resistance than human brain glioma TJ905 cells to temozolomide, which interpret the drug resistance machanism of glioma cells and stem cells.