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-  2015 

WNT6在BMSCs增殖、迁移和成骨分化中的作用

DOI: doi:10.7507/1002-1892.20150019

Keywords: WNT6, BMSCs, 细胞增殖, 细胞迁移, 成骨分化

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Abstract:

目的探讨WNT6对BMSCs增殖、成骨分化和迁移能力的影响。 方法取小鼠BMSCs培养至30%~50%融合时,分别行WNT6沉默及过表达观察。WNT6沉默实验分为3组,分别为转染WNT6特异性shRNA组(A1组)、转染对照shRNA组(B1组)、未转染的正常细胞组(C1组)。过表达观察实验分为3组,分别为转染WNT6重组表达质粒组(A2组)、转染空载体组(B2组)、未转染的正常细胞组(C2组)。倒置显微镜下观察细胞形态,A1、B1、A2、B2组筛选稳定转染细胞;于转染后48 h取细胞,采用实时荧光定量PCR检测WNT6 mRNA表达,Western blot检测WNT6及Ki73蛋白表达,Transwell法检测细胞迁移,MTT法检测细胞增殖;C1、C2组培养后相同时间点取细胞进行以上观测。各组细胞经成骨诱导培养12 d,检测ALP活性及钙沉积量。 结果倒置显微镜下,A1、B1、C1组及A2、B2、C2组细胞形态基本一致。A1组WNT6 mRNA及蛋白、Ki67蛋白表达、细胞增殖及细胞迁移数目、ALP活性以及钙沉积量均显著低于B1、C1组,差异均有统计学意义(P<0.05),B1、C1组间比较差异均无统计学意义(P>0.05);而A2组以上各指标均显著高于B2、C2组,差异均有统计学意义(P<0.05),B2、C2组间比较差异均无统计学意义(P>0.05)。 结论WNT6能促进小鼠BMSCs增殖和迁移,并增强其成骨分化能力

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