重组腺病毒AdKDR-TK联合IL-12逆转录病毒血管靶向治疗裸鼠皮下人肝癌细胞移植瘤的实验研究

GENE THERAPY WITH RECOMBINANT ADENOVIRUS ADKDR-TK AND IL-12 RECOMBINANT RETROVIRUS IN VESSELTARGETEDTREATMENT OF SUBCUTANEOUSLY IMPLANTED HHC IN NUDE MICE

目的探讨重组腺病毒AdKDR-TK联合IL-12逆转录病毒通过血管靶向治疗裸鼠皮下人肝癌细胞移植瘤的疗效及作用机制。方法 SCID小鼠腋部皮下植入1×107(0.2mL)HepG2细胞,同时腹腔注射PBL2×107(0.5mL),当瘤体达0.5cm3时,随机分成4组,分别为更昔洛韦(GCV)组(第I组)、IL-12逆转录病毒组(第Ⅱ组)、重组腺病毒AdKDR-TK组(第Ⅲ组)及重组腺病AdKDR-TK联合IL-12逆转录病毒组(第Ⅳ组)。各治疗组瘤内分别注入重组病毒液或/及重组逆转录病毒液0.1mL,第二天重复注射一次,重组腺病毒治疗在病毒给予24h后分别在腹腔内注射GCV,连续l0d;对照组腹腔内注入GCV。观察各组瘤体生长情况及免疫组化法测定肿瘤微血管密度(MVD)。结果第Ⅳ组经治疗后肿瘤的生长受到明显的抑制,其抑瘤率为62.14%,与第II组及Ⅲ组比较,后两者的抑瘤率分别为18.32%和32.73%(与第IV组比较,两者P<0.01)。肿瘤组织内的MVD,第I组为(40.1±6.7)个/mm2、第II组为(32.2±7.3)个/mm2、第III组为(27.6±7.1)个/mm2、第Ⅳ组为(7.7±4.1)个/mm2,其中第Ⅲ组与第Ⅱ组(P<0.05)、第Ⅳ组与第Ⅱ组(P<0.01)、第Ⅳ组与第Ⅲ组(P<0.01)之问的肿瘤内MVD比较均有统计学差异。结论 IL-12逆转录病毒能够增强重组腙病毒介导以KDR为启动子的胸苷激酶系统的血管靶向性地有效抑制肿瘤的生长。

Objective：To investigate the therapeutic efficacy of Recombinant adenovirus AdKDR-tk and mIL-12 recombinant retrovirus in vessel-targeted treatment of subcutaneously implanted HHC in nude mice.Methods：Thirty-two SCID mice were subcutaneously transplanted with human hepatocellular carcinoma and the tumors were allowed to grow till the volume reached 0.5cm3, then the mice were divided into 4 groups：Ganciclovir group（I）, mIL-12 recombinant retrovirus group（II）, AdKDR-TK＋group GCV（Ⅲ） and AdKDR-TK＋mIL-12 recombinant retrovirus＋GCV group（IV）.Recombinant adenovirus or mIL-12 recombinant retrovirus were injected intra-tumorally in all mice.Ganciclovir（GCV）was given at a dose of 100 mg/（kg·d）（ip）on the following day of injection with recombinant adenovirus for 10 days, Mierovessel density（MVD）of the tumors was determined with the immunohistorchemical methods and the growth of the tumor was observed.Results：Compared with group IV, The tumor inhibitory rate in sroup IV was 62.14 %, The rates in group Ⅲ and group II were 32.73 % and 18.32%（P〈 0.01）.The inhibition rates in group IV were superior to that of group II.The mean MVD in group I, Ⅱ, Ⅲ and IV was 40.1 ± 6.7, 32.2 ± 7.3, 27.6 ± 7.1, and 7.7 ± 4.1（microvessels/mm^2）, respectively.Conclusion：The results demonstrated that pAdKDR-TK＋GCV system efficiently work together in the targeting treatment of vascular endothelial cells of HCC in vivo.And IL-12 could enhance the therapeutic effect.It may be a new treatment approach for human HCC.