微小RNA-203靶向抑制P63诱导人表皮干细胞向汗腺细胞分化

来源 :中国细胞生物学学会2015年全国学术大会 | 被引量 : 0次 | 上传用户:ferret
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  本研究观察微小RNA-203(miR-203)对人表皮干细胞表型转化的影响,探讨其诱导表皮干细胞向汗腺细胞分化的可能性及机制。取包皮环切术后的人正常包皮组织采用胰酶消化法分离表皮获得单细胞悬液,应用Ⅳ型胶原差速贴壁法分离培养人表皮干细胞。倒置显微镜下观察细胞的生长状况,免疫细胞化学染色法行β 1整合素(ITGB1)、CK19、CK1、CK10、CK18、 CEA单克隆抗体检测鉴定;根据人源miR-203序列,设计并合成其双链模拟物。通过脂质体转染将miR-203模拟物分子导入人表皮干细胞中,转染无关microRNA模拟物作为对照;免疫细胞化学染色法对转染后的2组细胞行ITGB1、CK19、CK1、CK10、CK18、CEA单克隆抗体检测鉴定;逆转录实时定量PCR技术检测2组细胞转染前后miR-203、P63、ITGB1、CK19、CK1、CK10、CK18、CEA基因表达量的变化;Western blot法检测2组细胞转染前后P63、ITGB1、CK19、CK1、CK10、CK18、CEA蛋白表达量的变化。
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