联合应用angiostatinK(1-3)和endostatin裸DNA对人脑胶质瘤生长的抑制作用

Inhibitive effects of combined application of eukaryotic vectors encoding angiostatinK (1-3) and endostatin gene on human gliomas growth

目的 :探讨联合应用angiostatinK(1 3) {ASK(1 3) }和endostatin(ES)裸DNA对人脑胶质瘤生长的影响和作用特点 .方法 :在 1 5只裸鼠皮下接种SHG4 4人脑胶质瘤细胞并分A ,B和C组 (n =5 ) ;在接种后第 1 2日 ,A组 :给予瘤体内注射 2 0 μL脂质体包裹的各 5 μg的 pcDNA S ASK (1 3)和pcDNA S ES质粒 ;B组 :瘤体内注射 2 0 μL脂质体包裹的 1 0μgpcDNA 3.1质粒 ;C组 :瘤体内注射 2 0 μL无菌生理盐水 ;3d后重复注射 1次 .每日定时观察肿瘤生长情况 .4 0d后处死裸鼠 ,检测和计算肿瘤体积、坏死率、血管数和抑瘤率 .结果 :A ,B和C组肿瘤的终体积分别是 4 .4 1 6± 1 .88cm3 ,8.0 34± 1 .85cm3 和 8.0 5 6± 1 .91cm3 ,A组抑瘤率为 4 6 .8% ;A ,B和C组肿瘤的平均坏死率分别为 39.1 2± 7.35 % ,9.1 7± 7.89%和 9.2 1± 7.4 8% ,A与B或C组比较差异显著 (P <0 .0 1 ) ,B ,C组比较差异不显著 (P >0 .0 5 ) .3组的血管计数分别为 3.92± 1 .89,1 1 .8± 2 .0 4和 1 2 .0± 2 .2 1 ,A与B或C组比较差异显著 (P <0 .0 1 ) ,B ,C组比较差异不显著 (P >0 .0 5 ) .结论 :联合应用ASK(1 3)和ES裸DNA可抑制胶质瘤的血管生成、导致其坏死增加 ,进而抑制肿瘤生长 .其作用机制有待深入研究 。

AIM: To discuss the character of the inhibitive effects of combined application of eukaryotic vectors encoding angiostatinK (1 3) and endostatin gene on the growth of human gliomas. METHODS: After SHG44 glioma models were established and divided into A, B and C groups at the 12th day, the vectors of pcDNA–S ASK(1 3), pcDNA S ES and normal saline of 20 μL (each with 5 μg DNA in total 20 μL liquid) packaged by lipofectamine were injected into the cells of rats in A, B and C groups. The nude DNA was injected separately at 3 day interval. The nude mice were put to death at the 40th day. The glioma growth was observed and the reduction of tumor size, tumor necrosis and micrangium were calculated. RESULTS: End tumor volumes of tumor cells in A, B and C groups were 4.416 cm 3, 8.034 cm 3 and 8.056 cm 3 respectively. The inhibitive rate of tumor cells in group A was 46.8%. The average necrosis rate of cells in A, B and C groups were 39.12%, 9.17% and 9.21% respectively ( P <0.01, group B or C compared with group A; P >0.05, group B compared with group C). The micrangium of cells in group A,B and C groups were 3.92±0.59, 11.8±2.04 and 12.0±2.21 respectively ( P <0.01, group B or C compared with group A; P >0.05, group B compared with group C). CONCLUSION: The human glioma growth was inhibited by the combined application of AK(1 3) and endostatin nude DNA. Its inhibitive effects may result from the tumor angiogenesis and tumor necrosis but the mechanism needs to be further explored. Our findings provide an elementary basis for the treatment of glioma by antiangiogenic method.