Gli2在髓母细胞瘤中的表达及其靶向抑制剂的作用与意义

摘要

Objective To investigate the expression of downstream transcription factor glioma -associated oncogene homologue 2 (Gli2) in sonic Hedgehog signaling pathway of medulloblastoma (MB) and its targeted inhibitors on MB cell proliferation effect .Meth-ods Doay cells were treated with different concentrations of GANT 61.Changes in cell morphology was observed and the effect of Gli 2 on proliferation of Doay cells was detected by cell counting Kit -8 (CCK-8) assay.RT-PCR and Western blot were used for detecting the expression of Gli2 mRNA and the level of Gli2 protein.Results Exposure of Doay cells to GANT61 in different concentrations showed significant changes in cell morphology and resulted in significant growth inhibition compared to normal control group (P<0.05).And the CCK-8 assay results showed that proliferation inhibition was in a dose -dependent manner.The expression of Gli2 mRNA was signifi-cantly decreased in the groups treated with GANT 61 compared to normal control group (P<0.05).Similarly, the expression of Gli2 pro-tein was significantly decreased in GANT61 group compared to normal group (P<0.05).Conclusion GANT61 inhibited the prolifera-tion of Doay cells mainly by down -regulating the expression of Gli 2 mRNA and its protein level .Therefore, Gli2 could be an important biological marker of MB and a potential biological therapeutic target .%目的 探讨SHH ( sonic hedgehog signaling pathway )信号通路下游转录因子Gli2 在髓母细胞瘤( medulloblastoma , MB)中的表达及其靶向抑制剂对MB细胞增殖抑制的作用. 方法 使用不同浓度的Gli2特异性抑制剂GANT61作用Doay细胞,观察细胞形态的变化,采用CCK-8法检测细胞的增殖抑制作用,RT-PCR法和Western blot 法分别检测Doay细胞中Gli2 mRNA的表达量及Gli2蛋白表达水平的差异. 结果 使用GANT61作用细胞后,可见细胞形态出现明显改变,细胞异常突起增多;CCK-8法检测显示,与对照组相比,GANT61能够抑制Doay细胞的增殖(P<0.05),且细胞增殖的抑制率呈明显的剂量-效应关系;RT-PCR法显示Gli2特异性抑制剂GANT61组较对照组Gli2 mRNA的表达明显下降(P<0.05);Western blot法检测结果示GANT61抑制剂组较对照组Gli2蛋白表达水平明显下降(P<0.05). 结论 GANT61主要通过抑制Gli2 mRNA的表达,下调Gli2蛋白表达水平而抑制MB的增殖生长. 因此,Gli2可能是治疗MB的重要生物学标志物和生物治疗的潜在靶点.