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 papilla cell,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ARRNA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.