【作者】 韩明勇； 刘奇； 唐步坚； 邓砚； 曹明峰；

【Author】 Han Ming-Yong Liu Qi Tang Bu-Jian Deng Yan Cao Ming-Feng Shan Dong People Hospital, Jinan 250117, Shandong Province. Guang Xi Medical University, Nanning 530021, Guangxi Province.

【机构】 山东省立医院肿瘤中心； 广西医科大学附属肿瘤医院；

【摘要】 目的:建立转人白细胞介素-18(hIL-18)基因的大肠癌细胞株SW480细胞,对IL—18基因修饰后的SW480 细胞进行灭活,制备大肠癌基因工程肿瘤疫苗,并对其抗肿瘤作用进行观察。方法:应用MTT法观察X线照射后对大肠癌细胞(SW480细胞)生长的影响血清细胞因子的检测, 实验鼠治疗15天后处死,取血检测血清IFN-γ、IL-2、IL- 12、IL-4、IL-10,采用ELISA法。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细胞子裸鼠右侧背部皮下,观察灭活后SW480- IL-18细胞的抗肿瘤效应。结果:经过高能X线照射后, SW480-IL-18细胞生长停滞,到第96小时,全部SWIL-18- 480细胞死亡。ELISA检测结果显示,未照射的106个SW48- IL-18细胞在24小时内分泌IL-18的含量是182．7 pg,照射的106个SW48-IL-18细胞在24小时内分泌IL-18的含量是 60．39 pg。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组与SW480组和 pcDNA3．1／SW480组比较(P<0．01),能明显刺激T细胞增殖,而SW480组和pcDNA3．1／SW480组比较对T细胞的刺激作用无明显差别(P>0．05),对T细胞的刺激作用微弱。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,治疗组肿瘤生长明显受到抑制,与其他对照组差别显著(P<0．05)。治疗组小鼠存活期显著延长,有33．35％小鼠长期存活,与其他对照组差别显著(P<0．05)。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。更多还原

【Abstract】 Objective: To study the effects of interleukin-18 gene transfection on tumorigenesis of human Colon carcinoma cell line SW480. Methods’-We transfected SW480 cells with a mammalin expression vector containing the human IL-18 complementary DNA. The biological expression of rhIL-18 was tested by RT-PCR and ELISA method, nude mice were injected with SW480 cells with or without hIL-18 gene. Results: The hIL-18 cDNA was successfully integrated into SW480 cells, it was 113.8 ± 4.7pg hIL-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-IL-18 in vivo was significantly slower than those of SW480 and SW480-pcDN A3.1 cells. Tumor formation by parental SW480 cells was significantly delayed in mice those had been immunized with SW480-IL18 irradiated cells.Conclusion: These findings indicated that human colon carcinoma cells were successfully modified by the gene of IL-18 cytokine and can reduce their tumorigenicity. Breast cells transduced with IL-18 gene might be used as human colon cancer vaccine.更多还原