下调RNA去甲基化酶FTO的表达对胶质母细胞瘤干细胞功能的影响

Effect of downregulation of RNA demethylase FTO expression on glioblastoma stem cell function

目的 分析下调N6-甲基酰胺(m6A)去甲基化酶脂肪组织和肥胖相关蛋白(FTO)的表达对人胶质母细胞瘤干细胞(GSC)增殖与干性维持的影响。方法 构建稳定下调FTO表达的干细胞株,运用CCK-8法检测下调FTO对GSC细胞增殖的影响;应用神经球形成实验检测两组细胞神经球形成能力的变化;运用体外有限稀释实验检测两组细胞自我更新能力的影响;流式细胞术检测下调FTO对GSC细胞凋亡的调控作用。结果 CCK-8结果显示,与对照组相比,下调FTO表达后减慢了GSC细胞生长速度。成球实验结果表明实验组的GSC神经球大小和数量较对照组显著减少。体外有限稀释实验结果显示,下调FTO表达后神经球自我更新能力减弱;流式细胞术检测凋亡显示,下调FTO表达后增加了GSC细胞凋亡率。结论 下调FTO的表达可抑制GSC的生长与自我更新能力,靶向FTO可能是清除GSC的潜在策略之一。

Objective To analyze the effects of down-regulated expression of m6A demethylase FTO on the proliferation and maintenance of glioblastoma stem cells(GSCs).Methods FTO knock-down glioma stem cells were generated to detect the effect of down-regulated FTO on GSC cell proliferation.The neurosphere formation ability of two groups of GSC cells was assessed by neurosphere formation experiment.In vitro limited dilution assays were used to examine the effects on self-renewal ability of GSC cells after FTO knockdown.Flow cytometry was used to detect the effect of down-regulated FTO on apoptosis of GSC cells.Results The CCK-8 results showed that compared with the control group,downregulation of FTO expression slowed down the growth rate of GSC cells.The results of the ball formation experiment showed that the size and number of GSC nerve balls in the experimental group were significantly reduced compared to the control group.The results of the in vitro limited dilution experiment showed that the self-renewal ability of the neurosphere was weakened after downregulating FTO expression.Flow cytometry detection of apoptosis showed that downregulation of FTO expression increased the apoptosis rate of GSC cells.Conclusion Downregulating the expression of FTO can inhibit the growth and self-renewal ability of GSC,and targeting FTO may be one of the potential strategies to clear GSC.