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目的:以PEI-壳聚糖(CP)为载体材料,考察CD133~+呈梯度表达的4种结肠癌细胞上的转染效率,将特异性结合CD133的TR多肽修饰CP材料,探讨材料与转染的关系。方法:根据CD133~+含量将细胞分为高表达组和低表达组,合成法制备CP材料,激光粒度仪测定纳米粒粒径、电位,CCK8实验检测细胞毒性,用质粒p GL3和EGFP考察CP材料的转染效果,Western印记检测CD133~+蛋白的表达。结果:CD133~+高表达组SW480、HCT116的CD133~+含量为96.97%、92.96%,低表达组CaCO_2、SW620的CD133~+含量为2.00%、0.56%;CCK8结果得出,CP材料按质量比N/P=5时,生物安全性较高;质粒p GL3转染高表达组,其CD133~+RLU值低于低表达组;质粒EGFP转染高表达组,绿色荧光弱于低表达组,Western blot检测接TR肽的CP材料CD133~+表达高于CP材料。结论:CP材料在低表达CD133~+组转染效率高,且接TR肽的CP材料针对CD133~+有可能介导材料进入细胞。
OBJECTIVE: To investigate the transfection efficiency of PEI-chitosan (CP) on four colon cancer cell lines with gradient expression of CD133 ~ +, CP material that specifically binds to CD133 was modified to CP material, Dyed relationship. Methods: The cells were divided into high expression group and low expression group according to the content of CD133 +. CP material was prepared by synthetic method. The particle size and potential of nanoparticles were measured by laser particle sizer. The cytotoxicity was detected by CCK8 assay. The transfection efficiency of the material was tested by Western blot to detect the expression of CD133 ~ + protein. Results: The CD133 ~ + content of SW480 and HCT116 was 96.97% and 92.96% respectively in CD133 ~ + high expression group, while the CD133 ~ + content of CaCO_2 and SW620 in low expression group was 2.00% and 0.56% The biosafety was higher than N / P = 5. The expression of CD133 ~ + RLU in plasmid pGL3 was lower than that in the low expression group. The plasmid EGFP was transfected into the high expression group, and the green fluorescence was weaker than the low expression group The expression of CD133 + in CP material detected by Western blot was higher than CP material. CONCLUSION: The transfection efficiency of CP material in the low expression of CD133 ~ + group is high, and the CP material of TR peptide targeting CD133 ~ + may mediate the entry of the material into the cell.