摘要
目的:制备人白细胞介素-18(hIL-18)基因的修饰的大肠癌基因工程肿瘤疫苗,观察其抗肿瘤作用。方法:应用MTT法观察X线照射后对SW480细胞生长的影响,治疗15d后,采用ELISA法检测实验鼠血清IFN-γ、IL-2、IL-12、IL-4、IL-10;将SW480细胞和SW480-IL-18细胞应用放射线灭活后接种于裸鼠左侧背部皮下,再接种SW480细胞于裸鼠右侧背部皮下,观察灭活后SW480-IL-18细胞的抗瘤效应。结果:经过高能X线照射后,SW480-IL-18细胞生长停滞,96h细胞全部死亡。未照射的106个SW48-IL-18细胞在24h内分泌IL-18的含量是182.7PG;照射组可是60.39pg。空载体转染的SW480细胞和未转染的SW480细胞上清液中未检测到IL-18。SW480-IL-18组较SW480组和pcDNA3.1/SW480组,能明显刺激T细胞增殖(P=0.01);而SW480组和pcDNA3.1/SW480组,对T细胞的刺激作用微(P=0.27)。灭活SW480-IL-18细胞能有效地激活裸小鼠体内NK细胞活性。SW480-IL-18细胞治疗后,治疗组较其他对照组肿瘤生长明显受到抑制(P<0.05)。治疗组小鼠存活期显著延长,有33.4%小鼠长期存活,高于对照组。结论:IL-18基因修饰的SW480细胞具有明显的抗肿瘤作用,为大肠癌基因工程肿瘤疫苗的研制提供了实验基础。
Objective：To study the effects of interleukin-18 gene transfection on tumorigenesis of human colon carcinoma cell line SW480.Metheds： We transfected SW480 cells with a mammalin expression vector containing the human IL-18 complementary DNA. The biological ex- pression of rhlL-18 was tested by RT-PCR and ELISA method; nude mice were injected with SW480 cells with or without hlL-18 gene. Resulte： The hlL-18 cDNA was successfully integrated into SW480 cells, it was 113.8 ± 4.7.hlL-18 secreted by one million transduced cells in 24 hours. In vivo tumor growth curves, based on measured tumor size, SW480, SW480-pcDNA3.1 and SW480-IL-18 resulted in tumor formation 8-11 days after injection. The growth rate of SW480-1L- 18 in vivo was significantly slower than those of SW480 and SW480-pcDNA3.1 cells. Tumor formation by parental SW480 cells was significantly delayed in mice those had been immunized with IL-18 irradiated cells. Couclusion： These findings indicated that human colon carcinoma cells were successfully modified by the gene of IL-18 cytokine and can reduce their tumorigenicity, colon cells transduced with IL-18 gene might be used as human cancer vaccine.
出处
《中国现代普通外科进展》
CAS
2007年第1期29-32,36,共5页
Chinese Journal of Current Advances in General Surgery
基金
山东省重点科研基金资助项目(Z2003CO1)
