Benimetskaya L, Takle G B, Vilenchik M, Lebedeva I, Miller P, Stein C A
Department of Medicine and Department of Pharmacology, Columbia University, 630 West 168th Street, New York, NY 10032, USA.
Nucleic Acids Res. 1998 Dec 1;26(23):5310-7. doi: 10.1093/nar/26.23.5310.
Cationic porphyrins form stable complexes with oligodeoxynucleotides. To evaluate delivery, we used a 20mer phosphorothioate oligomer (Isis 3521) targeted to the 3'-untranslated region of the PKC-alpha mRNA, and complexed it with porphyrin. The expression of PKC-alpha protein and mRNA in T24 bladder carcinoma cells was reduced by approximately 80 +/- 10% at a concentration of oligomer of 3 microM, and 9 microM porphyrin. The expression of PKC-beta1, -delta and -straightepsilon isoforms was unaffected by this treatment, but elimination of PKC-zeta protein and mRNA were observed. However, treatment with the porphyrin complex of Isis 3522, an oligomer which is directed at the 5' coding region of the PKC-alpha mRNA, was equally effective as Isis 3521 with respect to PKC-alpha, but did not affect PKC-zeta protein or mRNA levels. Since Isis 3521 has an 11-base region of complementarity with the PKC-zeta mRNA, wheras Isis 3522 has only a 4-base region, the effect of Isis 3521 on PKC-zeta protein and mRNA expression may be due to irrelevant cleavage. Depending upon the desired application, this new strategy may offer several advantages over other methods of antisense oligodeoxynucleotide delivery including efficiency, stability, solubility, relatively low toxicity and serum compatibility. Porphyrins may thus be a potentially useful delivery vehicle for antisense therapeutics and/or target validation.
阳离子卟啉能与寡脱氧核苷酸形成稳定的复合物。为评估递送效果，我们使用了一种靶向PKC-α mRNA 3'-非翻译区的20聚体硫代磷酸酯寡聚物（Isis 3521），并使其与卟啉复合。在寡聚物浓度为3 microM和卟啉浓度为9 microM时，T24膀胱癌细胞中PKC-α蛋白和mRNA的表达降低了约80±10%。PKC-β1、-δ和-ε亚型的表达不受此处理的影响，但观察到PKC-ζ蛋白和mRNA的消除。然而，用靶向PKC-α mRNA 5'编码区的寡聚物Isis 3522的卟啉复合物处理，在PKC-α方面与Isis 3521同样有效，但不影响PKC-ζ蛋白或mRNA水平。由于Isis 3521与PKC-ζ mRNA有一个11个碱基的互补区域，而Isis 3522只有一个4个碱基的区域，Isis 3521对PKC-ζ蛋白和mRNA表达的影响可能是由于无关的切割。根据所需的应用，这种新策略可能比其他反义寡脱氧核苷酸递送方法具有几个优势，包括效率、稳定性、溶解性、相对低毒性和血清相容性。因此，卟啉可能是反义治疗和/或靶点验证的一种潜在有用的递送载体。
