全营养液促脂质体介导质粒DNA转染结肠直肠癌细胞的实验研究

Experimental study of total nutrient admixture promoting transfection of plasmid DNA mediated with liposomes to colorectal cancer cells

目的 探讨全营养液 (TNA)促进脂质体介导质粒DNA转染结直肠癌细胞的作用。方法 采用细胞培养方法 ,利用绿色荧光蛋白基因做报告基因 ,配制不同转染液 :脂质体加pEGFP N1、TNA加脂质体加pEGFP N1、TNA加pEGFP N1、单纯脂质体、单纯TNA ,分别处理人结肠癌细胞LoVo、人直肠癌细胞HR 8348,检测癌细胞中绿色荧光蛋白的表达。 结果 未加pEGFP N1质粒DNA的单纯脂质体和单纯TNA处理的细胞不表达绿色荧光蛋白。TNA加脂质体加pEGFP N1对LoVo、HR 8348细胞的转染效率分别为 33%、38% ;脂质体加pEGFP N1组的转染效率为 2 2 %、2 4% ;TNA加pEGFP N1组的转染效率均为 1%。TNA加脂质体加pEGFP N1组转染效率高于其他转染组 (P <0 0 1)。质粒DNA对TNA溶液的稳定性无显著影响。 结论 TNA可促进脂质体介导质粒DNA转染结直肠癌细胞 ,将营养支持与基因治疗相结合 ,有望提高对肿瘤患者支持治疗效果。

Objective To study total nutrient admixture (TNA) promoting plasmid DNA transfection mediated with liposomes to colorectal cancer cells. Methods Dispensing varied transfection agents of liposome+DNA plasmid pEGFP-N 1, TNA+liposomes+pEGFP-N 1, TNA+pEGFP-N 1, liposomes merely, and TAN sole. Human colorectal cancer cell LoVo and HR-8348 were treated with the agents respectively. Green fluoresence protein (GFP) gene as a report gene was detected. Results GFP was not detected in cancer cells treated with agents of merely liposomes and TAN. Transfection rates of GFP in two groups of cancer cells treated with agent of TNA+liposomes+pEGFP-N 1 were 33%, 38% respectively. With liposome+pEGFP-N 1, the rates of transfection in two cells were 22%, 24% respectively. The expression of GFP was 1% in the two groups of tumor cells treated with TNA+pEGFP-N 1. With agent of TNA+liposomes+pEGFP-N 1, a high transfetion rate of GFP gene was obtained. And no negative effect was observed to stabilization of TAN solution. Conclusion TNA may enhance transfection rate of plasmid DNA mediated with liposome, and may be beneficial to the treatment of cancer.