调控Gpc1表达对神经胶质瘤细胞增殖的影响及其机制的初步研究

Hedgehog signaling pathway mediated Gpc1 regulation on the proliferation of neuroglioma cells

摘要:

目的：探讨调控Glypican1（Gpc1）表达对神经胶质瘤细胞株增殖能力的影响以及Hedgehog（Hh）通路在其中的作用，为神经胶质瘤的研究提供基础实验数据。方法：在４株人脑神经胶质瘤细胞中，采用Western blot挑选出Hh信号通路活性及Gpc1表达相对较高的细胞株，采用RNAi技术沉默Gpc1表达后，检测Hh信号通路关键分子Shh、Gli1和靶基因CyclinD1表达的变化，MTT法检测Gpc1 RNAi稳定细胞株增殖活力的改变；另构建Gpc1过表达质粒，检测对Hh信号通路活性改变。结果：采用RNAi沉默Gpc1表达后，Hh信号通路活性受到抑制（F=8 407.34，P=0.000），神经胶质瘤细胞株增殖活力下降（F=27.68，P=0.000）；上述结果可被Gpc1过表达所逆转。结论：Gpc1对神经胶质瘤细胞株的增殖能力有着明显的调节作用且该调节受Hh信号通路介导。

Abstract:

Objective：To explore the effects of altered expression of Gpc1 expression on the proliferation of glioma cells and to address the potential role of hedgehog signaling pathway in the mediation of Gpc1 action. Methods：One cell line having relatively high activa－tion in Hedgehog signaling pathway and higher Gpc1 expression from four glioma cell lines was selected via Western blot. Gpc1-RNAi stable cell line was constructed to detect the changes of Hedgehog pathway component. The activity of Hedgehog via luciferase reporter gene after altering Gpc1 expression was detected. Gpc1-RNAi stable cell line proliferation viability was detected by MTT. Results：The Hedgehog pathway was suppressed（F=8 407.34，P=0.000） and glioma cell viability was restricted（F=27.68，P=0.000） after knock－ing down Gpc1 expeession. Conclusion：Glioma cell proliferation can be regulated by Gpc1 via Hedgehog pathway