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该文主要研究了Hela细胞对阳离子脂质体/DNA复合体的摄取机制。通过使用抑制剂抑制真核细胞跨膜运输的相关路径,再通过阳离子脂质体将绿色荧光蛋白基因、荧光素酶报告基因转染到细胞中。利用荧光显微镜、微光检测仪定性以及定量检测不同抑制剂浓度下转染效率的差异,MTT法检测各浓度下的细胞存活率,确定对相应路径的依赖性。结果显示,在细胞存活率保持在60%以上的前提下,镜下观察到随着药物浓度的升高,荧光强度明显减弱,微光检测数据显示随着药物浓度增加,基因表达的效率具有明显的下降趋势。因此推测,Hela细胞主要通过网格蛋白以及小窝蛋白介导的路径摄取脂质体/DNA复合体,对于微管微丝介导的巨胞饮作用也具有一定的依赖性。
This paper mainly studied the mechanism of uptake of cationic liposomes / DNA complexes by Hela cells. The green fluorescent protein and luciferase reporter genes were transfected into the cells by cationic liposomes by using inhibitors to inhibit the transmembrane trafficking path of eukaryotic cells. Fluorescence microscopy, low light detector qualitative and quantitative detection of different concentrations of inhibitors of transfection efficiency differences, MTT assay cell viability at each concentration to determine the dependence on the corresponding path. The results showed that under the premise of keeping the cell survival rate above 60%, the fluorescence intensity decreased obviously with the increase of the drug concentration, and the microdosing detection data showed that with the increase of the drug concentration, the gene expression efficiency was obvious The downward trend. Therefore, we speculate that Hela cells uptake the liposome / DNA complex mainly through clathrin and caveolin-mediated pathways, which is also dependent on microtubule-mediated fibroblast growth.