Smyth A P, Rook S L, Detmar M, Robinson G S
Hybridon Incorporated, Worcester, Massachussetts 01605, U.S.A.
J Invest Dermatol. 1997 Apr;108(4):523-6. doi: 10.1111/1523-1747.ep12289740.
In psoriatic lesions, epidermal keratinocytes overexpress vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) and transforming growth factor alpha (TGF-alpha). TGF-alpha has been shown to induce VEGF/VPF in normal human epidermal keratinocytes in vitro. By using a 19-mer antisense phosphorothioate oligodeoxynucleotide (PS-ODN) complementary to bases 6-24 relative to the translational start site of the VEGF/VPF mRNA, the control sense and mismatched PS-ODNs, we examined modulation of VEGF/VPF induction by TGF-alpha in vitro. Normal human epidermal keratinocytes were treated with PS-ODNs and Lipofectin for 8 h prior to the addition of TGF-alpha. Inhibition was assayed at the level of secreted protein by capture ELISA and mRNA expression was assayed by Northern blot analysis. The anti-sense PS-ODN was capable of inhibiting VEGF/VPF RNA and protein to near-basal levels. This inhibition was concentration dependent. No effect was observed with the sense or mismatch control PS-ODNs. These studies suggest that antisense oligonucleotide technology may be a potential therapy for the inhibition of angiogenesis associated with certain skin disorders such as psoriasis.
在银屑病皮损中，表皮角质形成细胞过度表达血管内皮生长因子/血管通透因子（VEGF/VPF）和转化生长因子α（TGF-α）。体外实验表明，TGF-α可诱导正常人表皮角质形成细胞表达VEGF/VPF。我们使用与VEGF/VPF mRNA翻译起始位点相对应的6-24位碱基互补的19聚体硫代磷酸反义寡脱氧核苷酸（PS-ODN）、对照正义寡核苷酸和错配PS-ODN，检测了TGF-α在体外对VEGF/VPF诱导的调节作用。在添加TGF-α之前，将正常人表皮角质形成细胞用PS-ODN和脂质体转染试剂处理8小时。通过捕获ELISA法在分泌蛋白水平检测抑制作用，通过Northern印迹分析法检测mRNA表达。反义PS-ODN能够将VEGF/VPF RNA和蛋白抑制至接近基础水平。这种抑制作用呈浓度依赖性。正义或错配对照PS-ODN未观察到效果。这些研究提示，反义寡核苷酸技术可能是抑制与某些皮肤疾病（如银屑病）相关的血管生成的一种潜在治疗方法。
