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
吴佳艳 1黄海燕 1宋宸宇 1蒋备战1
作者信息
- 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)
- 折叠
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.
Abstract
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 aliz-arin 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,miner-alized 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Key words
Versican/mouse dental papilla cell/cell proliferation/cell migration/odontogenic differentiation引用本文复制引用
基金项目
上海市卫生健康委科研项目(202140357)
上海申康医院发展中心新兴前沿技术联合攻关项目(SHDC12023115)
上海申康医院发展中心新兴前沿技术联合攻关项目(SHDC12022120)
出版年
2024
NETL
NSTL
万方数据