雄激素受体RNA干扰质粒的构建及其对毛乳头细胞雄激素受体表达的抑制作用研究

Construction of RNA interference plasmid targeting to androgen receptor gene and its effects on expression of androgen receptor in human dermal papilla cell

目的:构建雄激素受体(androgen receptor,AR)RNA干扰的真核表达质粒,并将其转染人毛乳头细胞(dermal papillacell,DPC)后观测其对AR表达的影响。方法:应用基因重组技术构建AR的RNA干扰质粒,经测序鉴定后以脂质体转染方法将质粒转染DPC,Western blot检测被转染细胞AR蛋白的表达。结果:成功构建了雄激素受体RNA干扰的真核表达质粒,将其转染了人DPC后,经Western blot证实AR RNA干扰质粒能使DPC细胞AR蛋白含量显著下降(P<0.05)。结论:AR的RNA干扰质粒能抑制DPC内AR的表达,该实验为雄激素性脱发基因治疗的实验研究提供了必要的基础。

Objective： To construct RNA interference （RNAi） expression vector targeting to androgen receptor （AR） gene and investigate its effects on the AR expression in human dermal papilla cell （DPC） after transfection. Methods： The RNAi plasmid vectors were constructed by using gene recombination technique, after selecting AR-specific siRNAs. Then, the DPCs were transfected by the recombinant plasmids through lipofection. The protein expression of AR was detected by Western blot. Results： The eukaryotic expression vector RNAi-AR was successfully constructed, the AR expression in DPCs was decreased after the transfection, shown by Western blot（P〈0.05）. Conclusion： The results suggest that RNA interference could inhibit the AR expression in DPCs, which is essential for the experimental gene therapy of androgenic alopecia.