Wnt-3a信号途径对毛乳头细胞聚集性生长作用的初步研究

Effect of Wnt-3a signaling on aggregative growth of dermal papilla cells： a pilot study

目的探讨Wnt-3a信号途径对体外培养的毛乳头细胞聚集性生长特性的影响。方法构建重组腺病毒pAdEasy-1／Wnt-3a，并以该重组腺病毒感染体外培养的毛乳头细胞，观察细胞生长特性的改变；应用激光共聚焦技术和Western印迹检测β-连环素及相关蛋白多功能蛋白聚糖的表达情况。结果Wnt-3a的过表达可以有效诱导毛乳头细胞聚集性生长。激光共聚焦显微镜检查提示，Wnt～3a过表达的毛乳头细胞β-连环素和多功能蛋白聚糖的荧光强度分别由34．16±9．62和18．81±9．59升高至87．35±17．28和92．18±18．39（t值分别为11．98，17．88，P值均〈0．01）；Western印迹也提示，Wnt-3a过表达的毛乳头细胞β-连环素和多功能蛋白聚糖的表达强度由15．27±2．17和19．32±2．46升高至60．09±7．24和58．46±2．78（t值分别为10．10，20．63，P值均〈0．01）。结论Wnt-3a是调节毛乳头细胞聚集性生长的重要分子，主要通过以β-连环素为核心蛋白的经典途径发挥调节作用，多功能蛋白聚糖作为Wnt信号途径的重要相关基因，是该调节的重要环节。

Objective To investigate the influence of Wnt-3a signaling on aggregative growth of dermal papilla cells. Methods Recombinant adenovirus pAdEasy-1/Wnt-3a was constructed at first, and used to transfect cultured dermal papilla cells （DPCs）. Then, the growth pattern of DPCs was observed by inverse microscopy; laser confocal microscopy and Western blot were utilized to detect the expressions of Wnt-3a, β-catenin and versican in low-passage DPCs, high-passage DPCs and transfected high-passage DPCs. Results The over-expression of Wnt-3a protein could effectively induce the aggregative growth of DPCs. Laser confocal microscopy showed that the fluorescence intensity of β-catenin and versican increased from 34.16± 9.62 and 18.81 ± 9.59 in high-passage DPCs to 87.35 ± 17.28 and 92.18 ± 18.39 in transfected high-passage DPCs （t = 11.98, 17.88, both P 〈 0.01 ）, respectively. Also, Western blot proved a significant increase of expression intensity of β-catenin （15.27 ± 2.17 vs 60.09 ± 7.24, t = 10.10, P 〈 0.01） and versican （19.32 ± 2.46 vs 58.46 ± 2.78, t = 20.63, P 〈 0.01） in transfected high-passage DPCs compared with untransfected high-passage DPCs. Conclusions Wnt-3a signaling plays an important role in modulating aggregative growth of DPCs, which is mainly through the classical pathway with β-catenin as the key protein. As an important downstream gene of Wnt signaling, vesican is an indispensable part for the modulation of aggregative growth of DPCs.