Transient activation of hedgehog pathway rescued irradiation-induced hyposalivation by preserving salivary stem/progenitor cells and parasympathetic innervation.
Transient activation of hedgehog pathway rescued irradiation-induced hyposalivation by preserving salivary stem/progenitor cells and parasympathetic innervation.
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DOI：
10.1158/1078-0432.ccr-13-1434
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发表时间：
2014-01-01
期刊：
影响因子：
--
通讯作者：
Liu F
中科院分区：
文献类型：
--
作者：
Hai B;Qin L;Yang Z;Zhao Q;Shangguan L;Ti X;Zhao Y;Kim S;Rangaraj D;Liu F
研究短暂激活 Hedgehog 通路对挽救放疗引起的头颈癌幸存者唾液分泌不足的作用和机制。小鼠唾液腺和培养的人唾液上皮细胞接受单次15Gy剂量照射。 Hedgehog 通路在小鼠唾液腺中通过短期过表达 Sonic hump (Shh) 转基因或施用 Smoothened 激动剂而被短暂激活，而在人唾液上皮细胞中则通过感染编码 Gli1 的腺病毒而被短暂激活。通过 Ptch1-lacZ 报告基因和内源 Hedgehog 靶基因的表达来检查 Hedgehog 信号传导的活性。在毛果芸香碱刺激后测量唾液流速。通过流式细胞术、唾液球测定、IHC、定量 RT-PCR、Western blot 和 ELISA 检查唾液干/祖细胞 (SSPC)、副交感神经支配和相关基因的表达。辐射不会激活小鼠唾液腺中的 Hedgehog 信号传导。短暂的Shh过表达激活了导管上皮中的Hedgehog通路，并且辐射后恢复了雄性小鼠的唾液功能，这与功能性SSPC的保存和副交感神经支配有关。 SSPC 的保存可能是通过挽救 Bmi1 和 Chrm1/HB-EGF 途径的信号活性来介导的。副交感神经支配的保存与神经营养因子（如 Bdnf 和 Nrtn）表达的恢复有关。雌性唾液腺和培养的人唾液上皮细胞中与唾液干/祖细胞维持和副交感神经支配相关的基因表达同样受到辐射和瞬时刺猬激活的影响。这些发现表明，Hedgehog 通路的瞬时激活有可能恢复辐射引起的唾液腺功能障碍。
To examine effects and mechanisms of transient activation of Hedgehog pathway on rescuing radiotherapy-induced hyposalivation in head and neck cancer survivors. Mouse salivary glands and cultured human salivary epithelial cells were irradiated by single 15Gy dose. Hedgehog pathway was transiently activated in mouse salivary glands by shortly over-expressing Sonic hedgehog (Shh) transgene or administrating Smoothened Agonist and in human salivary epithelial cells by infecting with adenovirus encoding Gli1. Activity of Hedgehog signaling was examined by expression of Ptch1-lacZ reporter and endogenous Hedgehog target genes. Salivary flow rate was measured following pilocarpine stimulation. Salivary stem/progenitor cells (SSPCs), parasympathetic innervation and expression of related genes were examined by flow cytometry, salisphere assay, IHC, quantitative RT-PCR, Western blot and ELISA. Irradiation does not activate Hedgehog signaling in mouse salivary glands. Transient Shh over-expression activated Hedgehog pathway in ductal epithelia and that after irradiation rescued salivary function in male mice, which is related with preservation of functional SSPCs and parasympathetic innervation. The preservation of SSPCs was likely mediated by rescue of signaling activities of Bmi1 and Chrm1/HB-EGF pathways. The preservation of parasympathetic innervation was related with rescue of expression of neurotrophic factors such as Bdnf and Nrtn. The expression of genes related with maintenance of salivary stem/progenitor cells and parasympathetic innervation in female salivary glands and cultured human salivary epithelial cells was similarly affected by irradiation and transient Hedgehog activation. These findings suggest that transient activation of Hedgehog pathway has the potential to restore irradiation-induced salivary gland dysfunction.