小GTP酶Arl6调控原纤毛生成和Shh信号转导
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国家自然科学基金青年基金项目(81101497);国家重点基础研究发展计划(973计划)(2012CB945003)


Small GTPase Arl6 regulates ciliogenesis and sonic hedgehog signal transduction
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    摘要:

    目的:研究小GTP酶Arl6对sonic hedgehog(Shh)信号通路的调控作用-方法:构建Arl6敲低稳转细胞株,检测Shh通路靶基因Gli1及Ptch1的mRNA表达水平;激光共聚焦显微镜下观察Arl6在原纤毛的定位,并且检测Arl6敲低细胞中原纤毛的生成情况-结果:Arl6的敲低抑制Shh信号通路的完全激活,在高浓度Shh条件性培养基或Smo激动剂Purm刺激下,靶基因Gli1 mRNA表达水平明显低于对照shControl组(P < 0.01),Ptch1 mRNA的表达水平也明显降低(P < 0.05);Arl6定位于原纤毛的基部小体,Arl6的缺失会影响原纤毛的生成;同时,Shh信号会刺激Arl6的表达(P < 0.05)-结论:小GTP酶Arl6的敲低抑制Shh通路的完全活化,与其抑制原纤毛生成有关-

    Abstract:

    Objective:To investigate the mechanism that small GTPase Arl6 regulates sonic hedgehog(Shh) signal transduction. Methods:The mRNA expression level of Gli1 and Ptch1,two Shh pathway target genes,were detected in Arl6 knockdown stable cell lines. The localization of Arl6 in the primary cilia was observed under confocal microscope. Ciliogenesis of Arl6 knockdown cells was quantified by counting the percentage of ciliated cells. Results:Knockdown of Arl6 inhibited full activation of the Shh pathway. After treatment with high-concentration Shh medium or Smo agonist Purm,Gli1 mRNA expression was significantly lower in the shArl6 group than in the control group (P < 0.01),and Ptch1 mRNA expression was also decreased (P < 0.05). Arl6 was located in the basal body of the primary cilia, and its absence resulted in defective ciliogenesis. Meanwhile,Shh signaling stimulated the expression of Arl6 (P < 0.05). Conclusion:Knockdown of small GTPase Arl6 inhibits full activation of Shh signaling,which is related to defective ciliogenesis.

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沈秋红,汤 颖,乐 珅,程 雁.小GTP酶Arl6调控原纤毛生成和Shh信号转导[J].南京医科大学学报(自然科学版),2014,(7):853-857

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  • 收稿日期:2014-03-25
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  • 在线发布日期: 2014-07-05
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