LncRNA-XIST promotes dermal papilla induced hair follicle regeneration by targeting miR-424 to activate hedgehog signaling
LncRNA-XIST promotes dermal papilla induced hair follicle regeneration by targeting miR-424 to activate hedgehog signaling
复制标题
LncRNA-XIST 通过靶向 miR-424 激活刺猬信号促进真皮乳头诱导的毛囊再生
DOI：
10.1016/j.cellsig.2020.109623
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发表时间：
2020-08-01
影响因子：
4.8
通讯作者：
Yin, Guo-Qian
中科院分区：
文献类型：
--
作者：
Lin, Bo-Jie;Zhu, Jiang-Ying;Yin, Guo-Qian
背景：脱发是一种以脱发为特征的高度流行的疾病。毛乳头(DP)细胞是毛囊再生的诱导剂，体外三维培养的DP细胞已被证明可以诱导毛囊再生。然而，3D培养的DP细胞的调控机制尚不清楚。方法：以3D培养的DP细胞作为体外细胞模型。将DP球移植到裸鼠体内进行毛囊再生的研究。QRT-PCR、Western blotting和免疫荧光法分别检测XIST、miR-424和Hedgehog通路相关蛋白的表达水平。用H&E染色观察毛发新生情况。采用四甲基偶氮唑盐比色法、CCK-8比色法和ELISA法分别检测细胞存活率、细胞增殖和碱性磷酸酶活性。结果：Xist和Shh在3D-DP细胞中表达上调，miR-424表达下调。分子调控研究表明，XIST海绵miR-424促进Shh的表达。XIST通过抑制miR-424的表达，抑制DP细胞活性、细胞增殖、ALP活性和其他DP标志物的表达。XIST基因敲除通过靶向miR-424抑制Shh介导的Hedgehog信号传导。结论：XIST海绵可促进Shh的表达，从而激活Hedgehog信号转导通路，促进DP介导的毛囊再生。
Background: Alopecia is a highly prevalent disease characterizing by the loss of hair. Dermal papilla (DP) cells are the inducer of hair follicle regeneration, and in vitro three-dimensional (3D) culturing DP cells have been proven to induce hair follicle regeneration. However, the molecular mechanisms behind the regulation of 3D culturing DP cells remain unclear.Methods: 3D-cultivated DP cells were used as in vitro cell model. DP sphere xenograft to nude mice was performed for in vivo study of hair follicle regeneration. qRT-PCR, Western blotting, and immunofluorescence were used for detecting the level of XIST, miR-424 and Hedgehog pathway-related proteins, respectively. H&E staining was used to examine hair neogenesis. Cell viability, proliferation and ALP activity were measured by MTT, CCK-8 and ELISA assays, respectively. Luciferase assays were used for studying molecular regulation between XIST, miR-424 and Shh 3'UTR.Results: XIST and Shh were up-regulated, and miR-424 was down-regulated in 3D DP cells. Molecular regulation studies suggested that XIST sponged miR-424 to promote Shh expression. Knockdown of XIST suppressed DP cell activity, cell proliferation, ALP activity and the expression of other DP markers by sponging miR-424. Knockdown of XIST suppressed Shh mediated hedgehog signaling by targeting miR-424. Moreover, the knockdown of XIST inhibited DP sphere induced in vivo hair follicle regeneration and hair development.Conclusion: XIST sponges miR-424 to promote Shh expression, thereby activating hedgehog signaling and facilitating DP mediated hair follicle regeneration.