CATIONIC LIPID IS NOT REQUIRED FOR UPTAKE AND SELECTIVE INHIBITORY ACTIVITY OF ICAM-1 PHOSPHOROTHIOATE ANTISENSE OLIGONUCLEOTIDES IN KERATINOCYTES
CATIONIC LIPID IS NOT REQUIRED FOR UPTAKE AND SELECTIVE INHIBITORY ACTIVITY OF ICAM-1 PHOSPHOROTHIOATE ANTISENSE OLIGONUCLEOTIDES IN KERATINOCYTES
复制标题
DOI：
10.1111/1523-1747.ep12396876
复制
发表时间：
1994-10-01
影响因子：
6.5
通讯作者：
NICKOLOFF, BJ
中科院分区：
文献类型：
--
作者：
NESTLE, FO;MITRA, RS;NICKOLOFF, BJ
角化细胞间粘附分子-1 （ICAM-1）在介导T细胞在表皮室内的滞留中起重要作用。为了确定设计用于与各种ICAM-1 mRNA区域杂交的反义寡核苷酸是否可以选择性地影响γ干扰素（ifn - γ）作用后培养的角质细胞ICAM-1的表达，将细胞暴露于几种反义化合物中，无论是否存在阳离子脂质（脂素）。角质形成细胞迅速内化正义和反义化合物（在30-60分钟内），即使在没有脂凝素的情况下，大约30%的细胞具有阳性细胞核。在培养的成纤维细胞、平滑肌细胞或内皮细胞中，尽管在所有这些细胞类型中，细胞质内的细胞摄取总量显著增加，但在缺乏脂凝素的情况下，没有观察到这种核积累。利用细胞计数技术，ifn - γ诱导的ICAM-1表达通过含有脂脂素的反义化合物降低50%，不含脂脂素的反义化合物降低30%。这种抑制是特异性的，因为没有观察到HLA-DR或肿瘤坏死因子α受体表达的变化。Northern blot杂交研究证实，ICAM-1反义寡核苷酸选择性地显著抑制ICAM-1的表达。这些结果表明，这些反义化合物与角质形成细胞的相互作用不同于其他细胞类型，并为临床试验提供了体外基础，在临床试验中，表皮角质形成细胞可以选择性地减少（或消除）ICAM-1的表达，而不一定影响成纤维细胞、内皮细胞或平滑肌细胞等真皮细胞类型。
Keratinocyte intercellular adhesion molecule-1 (ICAM-1) is important in mediating retention of T cells within the epidermal compartment. To determine if antisense oligonucleotides designed to hybridize to various ICAM-1 mRNA regions could selectively influence cultured keratinocyte ICAM-1 expression following gamma interferon (IFN-gamma), cells were exposed to several antisense compounds, in the absence and presence of cationic lipid (lipofectin). Keratinocytes rapidly internalized sense and antisense compounds (within 30-60 min), even in the absence of lipofectin with approximately 30% of the cell possessing positive nuclei. Such nuclear accumulation was not observed in the absence of lipofectin in cultured fibroblasts, smooth muscle cells, or endothelial cells, even though total cellular uptake within the cytoplasm was significantly increased in all these cell types. Using how cytometry, IFN-gamma-inducible ICAM-1 expression was reduced 50% by antisense compounds with lipofectin, and by 30% without lipofectin. This inhibition was specific as no change was observed for HLA-DR or tumor necrosis factor-alpha receptor expression. Northern blot hybridization studies confirmed that ICAM-1 antisense oligonucleotides selectively and significantly inhibited ICAM-1 expression. These results suggest that such antisense compounds interact with keratinocytes differently than other cell types, and provide the in vitro basis for clinical trials in which reduction (or elimination) of ICAM-1 expression by epidermal keratinocytes could be selectively accomplished without necessarily influencing dermal cell types such as fibroblasts, endothelial cells, or smooth muscle cells.