沉默Versican V0/V1对小鼠牙乳头细胞生物学行为的影响  

Effects of Versican V0/V1 gene silencing on biological behaviors of mouse dental papilla cells

作　　者：吴佳艳 黄海燕 宋宸宇 蒋备战[1] WU Jiayan;HUANG Haiyan;SONG Chenyu;JIANG Beizhan(Department of Pediatric Dentistry,Stomatological Hospital and Dental School of Tongji University,Shanghai Engineering Research Center of Tooth Restoration and Regeneration,Shanghai 200072,China)

机构地区：[1]同济大学口腔医学院·附属口腔医院儿童口腔科,上海牙组织修复与再生工程技术研究中心,上海200072

出　　处：《口腔医学》2024年第5期349-355,共7页Stomatology

基　　金：上海市卫生健康委员会科研项目(202140357);上海申康医院发展中心新兴前沿技术联合攻关项目(SHDC12023115,SHDC12022120)。

摘　　要：目的探究多功能蛋白聚糖(Versican)V0/V1对小鼠牙乳头细胞(mouse dental papilla cells,mDPCs)生物学行为的影响。方法体外培养E16.5d胎鼠mDPCs,使用小干扰RNA(siRNA)沉默mDPCs的Versican V0/V1基因,通过qRT-PCR和免疫荧光染色验证沉默效率;通过EdU实验检测细胞增殖率,划痕实验及Transwell细胞迁移实验检测细胞迁移能力;碱性磷酸酶(ALP)染色和茜素红染色分别评估mDPCs的矿化情况,并利用qRT-PCR检测其成牙和矿化相关基因表达水平。结果siRNA转染后,与si-NC组相比,si-Versican V0/V1组mDPCs的增殖能力及迁移能力减弱(P<0.01),si-Versican V0/V1组碱性磷酸酶染色更深且矿化结节数量增加,si-Versican V0/V1组成牙和矿化相关的基因表达量增加(P<0.05)。结论沉默Versican V0/V1抑制mDPCs增殖和迁移,但促进mDPCs成牙分化和矿化。Objective To investigate the effect of Versican V0/V1 on biological behaviors of mouse dental papilla cells(mDPCs).Methods mDPCs were isolated from C57BL/6J mice at embryonic day 16.5(E16.5).A small interfering RNA(siRNA)constructed specifically for Versican V0/V1 was transfected into mDPCs.The silencing efficiency was verified by quantitative real-time fluorescence polymerase chain reaction(qRT-PCR)and immunofluorescence staining.The proliferation rate of mDPCs was determined using EdU assay;the migration ability of mDPCs was evaluated by scratch test and transwell assay.Alkaline phosphatase(ALP)staining and alizarin red staining were used to assess the mineralization capability of mDPCs.The molecules related to odontogenic differentiation and mineralization at mRNA levels were measured by qRT-PCR.Results After siRNA transfection,the mDPCs of si-Versican V0/V1 group showed weaker proliferation and migration abilities compared with si-NC group(P<0.01).An enhanced ALP staining intensity,mineralized nodule formation and up-regulations of the molecules related to odontogenic differentiation and mineralization at mRNA levels(P<0.05)were observed in the mDPCs of si-Versican V0/V1 group.Conclusion Versican V0/V1 silencing inhibits the proliferation and migration of mDPCs,but enhances the abilities of odontogenic differentiation and mineralization.

关 键 词：多功能蛋白聚糖 小鼠牙乳头细胞 细胞增殖 细胞迁移 成牙分化 

分 类 号：Q28[生物学—细胞生物学]