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设计针对端粒酶RNA组分的特异性核酶,研究该核酶对肝癌细胞裸鼠移植瘤生长的影响。构建含针对端粒酶RNA组分锤头状核酶基因的重组真核表达质粒pBBS212-RZ,以及建立人肝癌细胞株HepG2裸鼠移植瘤模型。将不同剂量的pBBS212-RZ用脂质体包裹后进行瘤体内多点注射,对照组注射生理盐水或空质粒载体pBBS212。连续注射21天后,测量移植瘤体积和重量,检测瘤组织端粒酶活性。抗端粒酶特异性核酶使瘤组织端粒酶活性下降(抑制率为72％),明显地抑制了肝癌细胞裸鼠移植瘤生长(抑制率为68％),且存在剂量依赖性。抗端粒酶特异性核酶作为一种有效的端粒酶抑制剂,可抑制肝癌细胞的生长,有望在肝癌基因治疗中发挥作用。
The specific ribozyme against telomerase RNA was designed to study the effect of ribozyme on the growth of transplanted hepatoma cells in nude mice. To construct a recombinant eukaryotic expression plasmid pBBS212-RZ containing telomerase RNA hammerhead ribozyme gene, and to establish a human hepatoma cell line HepG2 xenograft model in nude mice. After different doses of pBBS212-RZ were encapsulated by liposomes, multiple injection in the tumor was carried out, and the control group was injected with saline or empty vector pBBS212. After 21 days of continuous injection, the volume and weight of the transplanted tumor were measured, and the telomerase activity of the tumor tissue was detected. Telomerase-specific ribozyme decreased the telomerase activity in the tumor tissue (72% inhibition rate), and significantly inhibited the growth of the transplanted hepatocarcinoma cells in nude mice (68% inhibition rate) in a dose-dependent manner. As an effective telomerase inhibitor, telomerase-specific ribozyme can inhibit the growth of hepatoma cells and is expected to play a role in gene therapy of hepatocellular carcinoma.