为探讨细胞因子之间对肿瘤细胞修饰的协同效果及融合基因用于肿瘤细胞转染的可行性,我们采用PCR及柔性接头连接的方法,构建了含人IL6/IL2融合基因的逆转录病毒载体,将其转染小鼠B16黑色素瘤细胞,对基因转导后细胞的粘附及实验性肺转移能力进行了测定.实验结果表明,融合基因转导的细胞所分泌产生的融合蛋白具有IL-2和IL-6两种细胞因子活性.对肿瘤细胞体外粘附及转移能力的测定结果表明,经融合基因修饰的B16细胞对细胞外基质的主要成分Laminin及重组基底膜Matrigel的粘附得到抑制,同时伴有实验性转移能力降低.这一结果提示,融合基因转导可明显降低肿瘤细胞的转移能力,其机制可能与基因修饰细胞相关生物学特性的改变及其对机体的免疫刺激活性有关.

In order to examine the feasibility of cytokine fusion gene transfer into tumor cells, a retroviral vector (pLXSN) for human interleukin 6/interleukin 2 (IL6/IL2) fusion gene was constructed by using PCR and ligating the 2 genes with a synthesized flexible linker. The IL6/IL2 fused gene was introduced into B16 melanoma cell line mediated by retrovirus and the changes of adhesive and metastatic ability of fused gene-modified cells were detected. The B16-IL6/ IL2 cells showed efficient expression of both cytokines and decreased binding affinity with extracellular matrix (Laminin and Matrigel), accompanying with decreased experimental metastatic potentials. These data suggest that the mechanism involving the decreased metastases may relate with the changes of biological characteristics and immune stimulating activity of gene-modified cells.

本课题受国家博士后科学基金资助