Campain J A, Matassa A A, Felgner P L, Barnhart K M, Curiel D T, Harrison G S
University of Colorado Health Sciences Center, Denver 80262, USA.
Cancer Gene Ther. 1998 May-Jun;5(3):131-43.
Kaposi's sarcoma (KS) is the most frequent malignancy occurring in HIV-positive individuals. AIDS-KS is a more aggressive disease than the classical form, frequently having a rapid clinical course with numerous serious complications. Current systemic treatments for KS, such as chemotherapy and the administration of biological modifiers, are complicated by both the drug resistance of the tumor and the dose-limiting toxicity of the reagents. The relative accessibility of many KS lesions makes the disease a particularly attractive candidate for in vivo gene therapy protocols. In this regard, we are interested in delivering conditionally toxic suicide and/or antiangiogenic vectors to accomplish targeted cell death selectively in AIDS-KS cells. To this end, we examined both cationic lipid- and adenoviral-mediated DNA transfection methods. Using the firefly luciferase reporter gene, we optimized numerous variables known to be important in lipid-mediated DNA transfection, including lipid formulation, the amount of lipid and DNA, lipid/DNA ratio, and cell concentration. Under optimal transfection conditions, approximately 5-25% of KS cells expressed the introduced DNA sequences. Adenoviral-mediated DNA delivery was more efficient than lipid delivery in 4 of 5 primary KS cell lines. Two of the lines (RW248 and RW376) were transduced by adenovirus at frequencies approaching 100%; two cell lines (CVU-1 and RW80) gave efficiencies of 20-35%. Two immortalized KS cell lines (KS Y-1 and KS SLK) were poorly infected, giving a transduction efficiency of <5%. These findings demonstrate that gene transfer into AIDS-KS cells is feasible, and suggest that vector strategies may be permissive for translating gene therapy approaches for the disease.
卡波西肉瘤（KS）是HIV阳性个体中最常见的恶性肿瘤。艾滋病相关卡波西肉瘤（AIDS-KS）是一种比经典型更具侵袭性的疾病，临床病程通常较快，伴有众多严重并发症。目前针对KS的全身治疗，如化疗和生物调节剂的给药，因肿瘤的耐药性和试剂的剂量限制性毒性而变得复杂。许多KS病变相对易于接近，这使得该疾病成为体内基因治疗方案特别有吸引力的候选对象。在这方面，我们有兴趣递送条件性毒性自杀和/或抗血管生成载体，以在AIDS-KS细胞中选择性地实现靶向细胞死亡。为此，我们研究了阳离子脂质介导和腺病毒介导的DNA转染方法。使用萤火虫荧光素酶报告基因，我们优化了许多已知在脂质介导的DNA转染中很重要的变量，包括脂质配方、脂质和DNA的量、脂质/DNA比率以及细胞浓度。在最佳转染条件下，约5%-25%的KS细胞表达了导入的DNA序列。在5个原发性KS细胞系中的4个中，腺病毒介导的DNA递送比脂质递送更有效。其中两个细胞系（RW248和RW376）被腺病毒转导的频率接近100%；两个细胞系（CVU-1和RW80）的转导效率为20%-35%。两个永生化KS细胞系（KS Y-1和KS SLK）感染较差，转导效率<5%。这些发现表明将基因转移到AIDS-KS细胞中是可行的，并表明载体策略可能有助于将基因治疗方法应用于该疾病。
