›› 2015, Vol. 35 ›› Issue (5): 341-344.

• Basic Research • Previous Articles     Next Articles

Effects of ERα gene silencing by ERα-shRNA mediated with lentiviral vector on theodonto /osteogenic differentiation of stem cells from apical papilla

  

  • Received:2014-12-16 Revised:2015-02-13 Online:2015-05-28 Published:2015-05-13

Abstract: 目的 构建雌激素受体α(estrogen receptor α,ERα)短发夹RNA(short hairpin RNA,shRNA)慢病毒表达载体,探讨其对根尖牙乳头干细胞(stem cells from apical papilla,SCAPs)体外成牙/成骨向分化的影响。方法 根据shRNA设计原则,合成用于体内干扰ERα的两条shRNA序列,重组入慢病毒载体PLKO.1。将PLKO.1对照组与ERα-shRNA (shERα-1和shERα-2)实验组分别感染SCAPs,western blot检测ERα的表达及其对SCAPs成牙/成骨向分化的影响。结果 测序结果证实重组质粒构建成功;体外实验表明,SCAPs转染ERα-shRNA后ERα蛋白表达水平降低,其中shERα-2组对ERα的下调作用更明显,两组ERα-shRNA中核心结合蛋白因子2(runt-related transcription factor-2,RUNX2)、成骨细胞特异性转录因子(osterix,OSX)、牙本质涎蛋白(dentin sialoprotein,DSP)和骨钙素(osteocalcin,OCN)的表达均有不同程度下调,与对照组相比具有显著性差异(P<0.05)。结论 慢病毒介导的ERα-shRNA成功在体外抑制了目的基因的表达,并显著影响了根尖牙乳头干细胞的成牙/成骨向分化。

Key words: Short hairpin RNA, Lentiviral vector, Estrogen receptor α, Stem cells from apical papilla

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