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目的探讨新型阳离子磷酸胆碱聚合物(MPC30DEA70)负载短片断干扰RNA(siRNA)进行细胞转染的可行性以及生物学效应。方法 MTT法分析MPC30DEA70对HepG2.2.15细胞的毒性;设计特异性siRNA,以MPC30DEA70负载红色荧光标记的siRNA转染HepG2.2.15细胞,在荧光显微镜下观察该载体携带siRNA能否进入细胞中及在该细胞中的转染效率。在此基础之上,观察对荧光素酶基因表达的抑制效应。结果在MPC30DEA70的浓度为0.5、1、3、5μl时,未发现对HepG2.2.15细胞产生明显的毒性,细胞存活率均在80%以上,并且能够携带siRNA进入细胞内,在48h的转染效率达(52.33±1.97)%,转染后能够有效地抑制荧光素酶质粒在HepG2.2.15细胞内的表达。结论 MPC30DEA70携带的siRNA在HepG2.2.15细胞中有着较高的转染效率,并发挥一定的生物效应,有望成为有效的siRNA载体。
Objective To investigate the feasibility and biological effect of a novel cationic phosphocholine polymer (MPC30DEA70) loaded with short interfering RNA (siRNA) for cell transfection. Methods The cytotoxicity of MPC30DEA70 on HepG2.2.15 cells was analyzed by MTT assay. HepG2.2.15 cells were transfected with siRNA targeting MPC30DEA70 and transfected into HepG2.2.15 cells by fluorescence microscopy. Transfection efficiency in cells. Based on this, the inhibitory effect on luciferase gene expression was observed. Results At the concentrations of 0.5, 1, 3 and 5μl, no significant cytotoxity was found in HepG2.2.15 cells, and the viability of MPC30DEA70 was over 80%. The siRNA could enter into the cells at a 48h transfection efficiency (52.33 ± 1.97)%, which could effectively inhibit the expression of luciferase plasmid in HepG2.2.15 cells. CONCLUSION: The siRNA carried by MPC30DEA70 has high transfection efficiency in HepG2.2.15 cells and exerts certain biological effects, which is expected to be an effective siRNA vector.