Development of novel cationic liposomes for efficient gene transfer into peritoneal disseminated tumor
Development of novel cationic liposomes for efficient gene transfer into peritoneal disseminated tumor
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DOI：
10.1089/10430349950018346
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发表时间：
1999-04-10
影响因子：
4.2
通讯作者：
Kikuchi, H
中科院分区：
文献类型：
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作者：
Kikuchi, A;Aoki, Y;Kikuchi, H
通过体内筛选，发现一系列新型阳离子脂质可有效地将基因转移到腹膜播散性肿瘤中。含有二肉豆蔻酸的O，O ′-双十四酰基-N-（1，2-三甲基）氯化铵（DC-6-14）在体外显示出最高的转染活性，条件是存在10%胎牛血清。为了提高DC-6-14的转染效率，我们以各种比例添加二油酰磷脂酰乙醇胺（DOPE）和/或胆固醇（Chol）作为辅助脂质。阳离子脂质体含有DC-6-14，DOPE，和Chol的摩尔比为1：0.75：0.75和1：1：0.8保持有效的转染活性，在HRA，mEIIL和ES-2细胞系在体外含血清的条件下，通过荧光素酶测定。与我们的新的脂质体，转染效率更高的细胞增殖速度比细胞增殖速度较慢，这取决于有丝分裂活性所代表的标记指数。在mEIIL腹膜播散性肿瘤模型中，用lacZ基因特异性转染癌细胞。通过X-Gal染色不仅在腹水中的漂浮癌细胞中观察到基因转移，而且在腹膜播散的癌组织中也观察到基因转移。LacZ阳性细胞的百分比约为1%，显著高于市售Lipofectin（0.38%）、LipofectACE（0.62%）或LipofectAMINE（0.23%）。在mEIIL腹膜播散性肿瘤-裸鼠模型中，用我们的新型脂质体转移单纯疱疹胸苷激酶基因（HSV tie），然后用更昔洛韦（GCV）治疗，与对照小鼠相比，导致显著更长的存活期（p < 0.05，Cox-Mantel）。这些结果表明，这些脂质体显示出作为腹膜内播散性癌症患者的基因治疗工具的前景。
A novel series of cationic lipids has been found, by lit vivo screening, to be effective for gene transfer into peritoneal disseminated tumor. O,O' -Ditetradecanoyl-N-(Lu-trimethylammonioa chloride (DC-6-14), having dimyristyl acid, has shown the highest transfection activity in vitro, provided that 10% fetal bovine serum is present. To enhance the transfection efficiency of DC-6-14, we added dioleoylphosphatidylethanolamine (DOPE) and/or cholesterol (Chol) as helper lipids in various ratios. Cationic liposomes containing DC-6-14, DOPE, and Chol in molar ratios of 1:0.75:0.75 and 1:1:0.8 maintained efficient transfection activity under serum-containing conditions in HRA, mEIIL, and ES-2 cell lines in vitro, as determined by luciferase assay. With our novel liposomes, transfection efficiencies were higher in cells proliferating faster than in cells proliferating slower, depending on mitotic activity as represented by labeling index. In the mEIIL peritoneal disseminated tumor model, cancer cells were specifically transfected with the lacZ gene. Gene transfer was observed by X-Gal staining not only in floating cancer cells in the ascites, but also in the peritoneal disseminated cancer tissue. The percentage of LacZ-positive cells was about 1%, which was significantly higher than with commercially available Lipofectin (0.38%), LipofectACE (0.62%), or LipofectAMINE (0.23%). In the mEIIL peritoneal disseminated tumor-nude mouse model, herpes simplex thymidine kinase gene (HSV tie) transfer with our novel liposomes, followed by ganciclovir (GCV) treatment, resulted in significantly longer survival compared with control mice (p < 0.05, Cox-Mantel). These results suggest that these liposomes show promise as tools in gene therapy for patients with intraperitoneal disseminated cancer.