初级纤毛对人牙周膜细胞成骨相关基因表达的影响

Effect of Primary Cilia on Expression of Osteogenic-related Genes of Human Periodontal Ligament Cells

目的:通过沉默纤毛运输蛋白88(intraflagellar transport 88,IFT88)基因抑制人牙周膜细胞(human periodontal ligament cells,hPDLCs)中初级纤毛的形成,进一步研究其对hPDLCs成骨相关基因表达的影响。方法:实验分为对照组和shIFT88组。对照组感染无意序列,shIFT88组感染含IFT88基因有效短发卡RNA(short hairpin RNA,shRNA)的慢病毒。qRT-PCR、Western blot检测各组细胞中IFT88表达情况,免疫荧光检测初级纤毛形成,qRT-PCR检测成骨相关基因COL1A1,OCN,Runx2,BMP2表达量的变化。结果:与对照组相比,shIFT88组IFT88表达量明显下调(P<0.01),纤毛率降低(P<0.01),成骨相关基因COL1A,OCN,Runx2,BMP2基础表达量明显下调(P<0.01)。结论:沉默IFT88基因后,hPDLCs初级纤毛形成受阻,成骨相关基因表达量明显下调。

Objective：To study the expression of osteogenic-related genes of human periodontal ligament cells（hPDLCs）by knocking down IFT88 to inhibit the formation of primary cilia.Methods：Control shRNA and shIFT88 were infected into hPDLCs.The expression of IFT88 was detected by qRT-PCR and Western blot.The formation of primary cilia was detected by immunofluorescence and the expressions of osteogenic-related genes COL1 A1,OCN,Runx2,and BMP2 were detected by qRT-PCR.Results：The PKLO.1-shIFT88 was stably expressed in hPDLCs and inhibited the expression of IFT88 effectively（P〈0.01）.The formation of primary cilia was partly suppressed（P〈0.01）.The expressions of osteogenic-related genes like COL1 A1,OCN,Runx2,and BMP2 were decreased significantly（P〈0.01）.Conclusion：Knockdown IFT88 inhibited the formation of primary cilia and reduced the expressions of osteogenic-related genes of hPDLCs.