Mounkes L C, Zhong W, Cipres-Palacin G, Heath T D, Debs R J
California Pacific Medical Research Institute, San Francisco, California 94115, USA.
J Biol Chem. 1998 Oct 2;273(40):26164-70. doi: 10.1074/jbc.273.40.26164.
The factors controlling cationic liposome-DNA complex (CLDC)-based gene transfer in cells and in animals are poorly understood. We found that cell surface heparin/heparan sulfate-bearing proteoglycans mediate CLDC-based gene transfer and expression both in cultured cells and following intravenous gene delivery into animals. CLDC did not transfect Raji cells, which lack proteoglycans, but did efficiently transfect Raji cells stably transfected with the proteoglycan, syndecan-1. Fucoidan, heparin, or dextran sulfate, all of which are highly anionic polysaccharides, each blocked CLDC-mediated transfection both in cultured cells and following intravenous injection into mice, but had no effect on transfection by either recombinant adenovirus infection or electroporation. Intravenous pretreatment of mice with heparinases, which specifically cleave heparan sulfate molecules from cell surface proteoglycans, blocked intravenous, CLDC-mediated transfection in mice, confirming that proteoglycans mediate CLDC gene delivery in vivo. Modulation of proteoglycan expression may prove useful in controlling the efficiency of, as well as targeting the sites of, CLDC-based gene transfer in animals.
目前对于控制基于阳离子脂质体 - DNA复合物（CLDC）的基因在细胞和动物体内转移的因素了解甚少。我们发现，细胞表面带有肝素/硫酸乙酰肝素的蛋白聚糖在培养细胞中以及静脉内基因传递给动物后，介导基于CLDC的基因转移和表达。CLDC不能转染缺乏蛋白聚糖的Raji细胞，但能有效地转染稳定转染了蛋白聚糖syndecan - 1的Raji细胞。岩藻依聚糖、肝素或硫酸葡聚糖，这些都是高度阴离子化的多糖，它们在培养细胞中以及静脉注射到小鼠体内后，均能阻断CLDC介导的转染，但对重组腺病毒感染或电穿孔介导的转染没有影响。用能够特异性从细胞表面蛋白聚糖上切割硫酸乙酰肝素分子的肝素酶对小鼠进行静脉内预处理，可阻断小鼠体内静脉注射CLDC介导的转染，证实蛋白聚糖在体内介导CLDC基因传递。调节蛋白聚糖的表达可能有助于控制基于CLDC的基因在动物体内转移的效率以及靶向转移位点。
