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研究FUS1/hIL-12双基因阳离子脂质体复合物的生物学和药剂学特性,评估其作为肺癌基因治疗药物的转染效率、稳定性以及毒性。阳离子脂质体与质粒以不同质量比配制后用凝胶阻滞实验和激光粒度仪测定粒径和zeta电位确定阳离子脂质体与质粒DNA的最佳配比;通过分别转染增强型绿色荧光蛋白质粒EGFP和PVITO2-hIL12/FUS1双基因质粒入A549肺癌细胞,倒置荧光显微镜、细胞爬片免疫组化和酶联免疫法分别检测EGFP、FUS1和hIL-12在肺癌细胞中的表达;以琼脂糖凝胶电泳检测双基因脂质体复合物与血清和DNaseⅠ孵育的稳定性;通过双基因脂质体复合物与红细胞孵育后显微镜观察其对红细胞形态的影响。研究结果为将来FUS1/hIL-12双基因阳离子脂质体复合物用于肺癌的基因治疗奠定了基础。
To investigate the biological and pharmacological properties of the FUS1 / hIL-12 double gene cationic liposome complex and evaluate its transfection efficiency, stability and toxicity as a lung cancer gene therapy drug. Cationic liposomes and plasmids were prepared at different mass ratios. The optimal ratio of cationic liposomes to plasmid DNA was determined by gel retardation assay and laser particle sizer. Particle size and zeta potential were determined by transfecting enhanced green fluorescence EGFP and PVITO2-hIL12 / FUS1 double gene plasmids into A549 lung cancer cells. The expression of EGFP, FUS1 and hIL-12 in lung cancer cells were detected by inverted fluorescence microscope, cell slide immunohistochemistry and enzyme-linked immunosorbent assay. The stability of double gene liposome complex incubated with serum and DNase I was detected by sugar gel electrophoresis. The morphology of erythrocytes was observed under a microscope after incubation of erythrocytes with the bioliposome complex. The results laid the foundation for future gene therapy of lung cancer with FUS1 / hIL-12 double gene cationic liposome complex.