摘要
目的:探究配对相关同源框2(paired related homeobox 2,Prrx2)在小鼠磨牙胚不同发育时期的表达及其对磨牙胚发育的影响。方法:利用实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)分析配对相关同源框1(paired related homeobox 1,Prrx1)和Prrx2在小鼠胚胎12.5 d(E12.5)至E18.5,出生后1、3、7 d(P1、P3、P7)磨牙牙胚中的表达,以及其在E13.5磨牙、毛发、脑、心脏、结肠、肾、肺、唾液腺等组织中的相对表达情况;使用小干扰RNA(small interfering RNA,siRNA)沉默Prrx2基因的表达,通过对牙胚行体外器官培养,观察牙胚发育情况;并利用CCK8实验、细胞划痕实验观察牙胚间充质细胞的增殖和迁移能力。结果:RT-qPCR结果显示,相比其他组织,Prrx2在E13.5小鼠胚胎的磨牙、毛发、唾液腺中的表达量更高;相比于Prrx1,Prrx2磨牙胚发育后期(E16.5~E18.5)的表达水平仍高于初始阶段(E13.5);牙胚体外培养结果显示,沉默Prrx2后,牙胚宽度、高度及牙尖高度分别减少了(21.11±3.49)%、(12.08±5.30)%和(22.34±7.56)%(均P<0.05);CCK8实验、细胞划痕实验结果显示,沉默Prrx2后,牙胚间充质细胞增殖能力无变化(P>0.05)、迁移能力降低(P<0.05)。结论:Prrx2在牙胚形态成形的过程中,承担着重要的作用;Prrx2通过促进磨牙胚间充质细胞的迁移能力,进而影响磨牙胚生长和牙尖发育的进程。
Objective:To investigate the expression level of paired related homeobox 2(Prrx2)at different stages of mouse molar germ and its effect on molar germ development.Methods:Real-time quantitative polymerase chain reaction(RT-qPCR)was used to analyze the relative expression of paired related homeobox 1(Prrx1)and Prrx2 in E12.5 to E18.5,P1,P3,P7,and E13.5 molar germ,hair,brain,heart,colon,kidney,lung,salivary gland and other tissues;small interfering RNA(siRNA)was used to silence the expression of Prrx2,then tooth germ development was observed via vitro culture systems;CCK8 test and cell scratch test were utilized to observe the proliferation and migration ability of molar germ mesenchymal cells.Results:According to the results of RT-qPCR,comparing with Prrx2 in other organs,it was highly expressed in the molars,hair and salivary glands of E13.5 mice;comparing with Prrx1,the expression level of Prrx2 at the late stage of tooth germ development(E16.5-E18.5)is still higher than that at the initial stage of development(E13.5);the results of in vitro tooth germ culture demonstrated that after silencing Prrx2,tooth germ width,height and tooth cusp height were reduced by(21.11±3.49)%,(12.08±5.30)%and(22.34±7.56)%(all P<0.05),respectively;CCK8 test and cell scratch test showed that the proliferation rate of tooth germ mesenchymal cells did not change after silencing Prrx2(P>0.05),but the migration ability was suppressed(P<0.05).Conclusion:Prrx2 plays an important role in the process of molar germ morphogenesis;Prrx2 affects the growth of molar germ and the process of tooth cusp development via promoting the migration ability of molar germ mesenchymal cells.
作者
朱辙文
韩雪
刘力维
王佐林
ZHU Zhewen;HAN Xue;LIU Liwei;WANG Zuolin(Shanghai Engineering Research Center of Tooth Restoration and Regeneration,Department of Oral Implantology,School and Hospital of Stomatology,Tong ji University,Shanghai 200072,China)
出处
《口腔颌面外科杂志》
CAS
2023年第3期131-137,共7页
Journal of Oral and Maxillofacial Surgery
基金
国家自然科学基金(82000995)
国家重点研发计划(2018YFE0202200)
中央高校基本业务费专项资金(22120180196)。
