【摘要】 目的:建立转人白细胞介素-18(IL-18)基因的乳腺癌细胞株Bcap37细胞,并探讨IL-18基因转导后Bcap37细胞肿瘤原性的改变和抗瘤作用。方法:将携带人IL-18的质粒pcDNA3.1-IL-18转导入人乳腺癌细胞系Bcap37细胞中,通过药物G418进行筛选,采用RT-PCR和ELISA法对IL-18基因的表达进行检测;裸鼠致瘤实验观察肿瘤原性的改变;观察灭活后Bcap37-IL-18细胞的抗瘤作用。结果:IL-18基因成功转导入Bcap37细胞中并能顺利表达;RT-PCR电泳结果显示IL-18基因在mRNA水平有表达;ELISA结果显示,106个转导细胞在24 h内分泌IL-18的含量是(126.3±4.5)pg;空载体转染的细胞未检测到IL-18;裸鼠致瘤实验表明,接种Bcap37-IL-18细胞的裸鼠肿瘤的生长速度明显低于Bcap37组和Bcap37-pCDNA3.1组;抗瘤实验结果显示,放射灭活的Bcap37-pcDNA3.1细胞和Bcap37-IL-18细胞免疫接种裸鼠后,再接种Bcap37细胞于裸鼠左侧背部皮下。肿瘤长出的时间Bcap37-IL-18细胞免疫接种组比Bcap37-pCDNA3.1细胞免疫接种组明显延长,且肿瘤的体积Bcap37-IL-18细胞免疫接种组比Bcap37-pCDNA3.1细胞免疫接种组小。结论:IL-18基因能成功的整合到Bcap37细胞基因组中,且在转导的肿瘤细胞中持续表达;IL-18基因转染降低了Bcap37细胞的肿瘤原性;IL-18基因修饰的Bcap37细胞具有明显的抗肿瘤作用。更多还原

【Abstract】 Objective: To study the effect of the interleukin-18 gene transfection on tumorigenesis of the human breast carcinoma cell line Bcap37.Methods: Bcap37 cells were transfected with a mammalin expression vector containing human IL-18 complementary DNA.The biological expression of IL-18 was determined by the RT-PCR and ELISA methods.Nude mice were injected with Bcap37 cells with or without the IL-18 gene.Results: The IL-18 cDNA was successfully integrated into the Bcap37 cells.There were(113.8±4.7)pg IL-18 secreted by one million transduced cells in 24 hours.In vivo tumor growth curves,based on measured tumor size,Bcap37,Bcap37-pcDNA3.1 and Bcap37-IL-18 resulted in tumor formation from 8 to 11 days after injection.The growth rate of Bcap37-IL-18 in vivo was significantly slower than that of Bcap37 and Bcap37-pcDNA3.1 cells.Tumor formation by parental Bcap37 cells was significantly delayed in mice that had been immunized with Bcap37-IL18 irradiated cells.Conclusion: Human breast carcinoma cellswere successfully modified by IL-18 cytokine genes and reduced the tumorigenicity.Breast cells transduced with IL-18 genes might be used as human colon cancer vaccines.更多还原