Cyclopamine对LN229细胞Nurr1基因表达的影响
王迪, 陈艳, 刘菲菲, 冷水龙, 龙大宏, 洪乐鹏
中国临床解剖学杂志 ›› 2012, Vol. 30 ›› Issue (5) : 539-542.
Cyclopamine对LN229细胞Nurr1基因表达的影响
Effect of cyclopamine on Nurr1 gene expression in LN229 cells
目的 探讨环巴胺(Cyclopamine)对恶性胶质瘤细胞株LN229帕金森相关基因Nurr1表达的影响。 方法 在脂多糖(LPS)刺激LN229细胞的条件下，运用实时定量PCR检测SHH通路基因PTCH、Gli1及Nurr1基因 mRNA含量，Western Blotting检测LPS对LN229细胞Nurr1蛋白表达的影响。进一步在Cyclopamine阻断Sonic Hedgehog (SHH)信号通路的条件下，实时定量PCR及Western Blotting检测Nurr1的mRNA含量及其蛋白的表达及下游炎症因子IL-1β mRNA的含量。 结果 LPS刺激下LN229细胞SHH通路相关基因mRNA，及Nurr1 mRNA升高，Western Blot结果进一步证明LPS可以促进Nurr1蛋白表达。Cyclopamine阻断组与对照组比较，在转录水平和翻译水平明显促进了Nurr1基因表达升高，及下游炎症因子IL-1βmRNA的含量。 结论 Cyclopamine对LN229细胞Nurr1表达的影响可能与帕金森发病过程中胶质细胞的活化有关。
Objective To investigate the effect of cyclopamine on the expression of Parkinson-related gene Nurr1 in malignant glioma cell line LN229. Methods Upon LPS stimulation, real time PCR was used to detect contents of SHH relational gene PTCH, Gli1and Nurr1 mRNA in LN229 cell. Western blotting was used to detect the impact of LPS on Nurr1 protein expression. Furthermore, the contents of both of Nurr1 and the downstream Inflammation factor IL-1β were detected after blocking SHH Signaling pathway with cyclopamine. Results Upon the LPS stimulation, the relational gene mRNA and Nurr1mRNA in LN229 cell elevated. LPS significantly elevated the expression of nurr1 protein. Contrast with the control group, it was significantly promoted on the expression of Nurr1 gene and the downstream Inflammation factor IL-1β at the level of transcription and translation after cyclopamine blocking. Conclusions The effects of cyclopamine on the Nurr1 gene expression in LN229 cell may be related to the activation of glial cells during the pathogenesis process of Parkinson's disease.
Cyclopamine / LN229细胞 / Nurr1
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国家自然科学基金资助项目(30900728)；广州市科技局基金资助项目(2700751-C0231)；广东省医学科学技术基金资助项目(A2010228)；广州市属高校科研基金资助项目(10A175)
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