Fam83h 突变对小鼠成骨细胞增殖和迁移的影响

Effects of Fam83h Mutation on Proliferation and Migration of Mouse Osteoblasts

目的:探究Fam83h突变对小鼠成骨细胞(mouse osteoblasts,MOBs)增殖和迁移的影响以及其相关机制。方法:Micro-CT扫描分析5月龄Fam83h突变纯合子小鼠和野生型小鼠的全身骨;体外分离培养Fam83h突变纯合子小鼠和野生型小鼠的MOBs,利用CCK-8实验、划痕实验研究Fam83h突变对MOBs增殖和迁移的影响并测序分析细胞转录组差异基因,Western blot检测相关通路关键分子表达情况。结果:Fam83h突变纯合子小鼠全身骨长及骨体积分数均小于野生型小鼠(P<0.001)。体外实验结果显示,Fam83h突变纯合子小鼠MOBs的增殖和迁移能力均显著低于野生型小鼠(P<0.01),KEGG Pathway富集分析提示PI3K/AKT通路差异明显,Western blot结果显示Fam83h突变纯合子小鼠的MOBs的p-AKT1-S473的蛋白水平降低。结论:Fam83h突变可能通过PI3K/AKT通路抑制MOBs的增殖和迁移从而导致小鼠骨量减少。

Objective:To explore the effect of Fam83h mutation on proliferation and migration of mouse osteoblasts(MOB)and the related mechanism.Methods:The whole body of wild-type and Fam83h-mutated mice were analyzed by micro-CT.CCK-8 and wound healing were detected in MOBs isolated from 3-day-old wild-type and Fam83h-mutated mice.RNA-seq was performed to compare the transcriptional profile of MOBs of Fam83h-mutated mice with wild-type mice.Phosphorylation of AKT1-S473 and AKT1 were measured by Western blot.Results:Fam83h-mutated mice showed decreased bone volume fraction(P<0.001).Fam83h mutation significantly repressed cell proliferation and migration in MOBs(P<0.01).KEGG pathway analysis of differentially expressed genes(DEGs)identified PI3K/AKT signaling pathway.Furthermore,the protein expression level of p-AKT1-S473 was decreased in MOBs of Fam83h mutation mice.Conclusion:Fam83h mutation may inhibit the proliferation and migration of MOBs through PI3K/AKT signaling pathway,resulting in bone mass loss in Fam83h-mutated mice.