人白细胞介素-16真核表达载体的构建及其对Th2型肿瘤细胞的促逆转作用

构建人IL-16真核表达载体，分析其对Th2型肿瘤细胞的逆转作用。方法用RT-PCR技术从人外周血单个核细胞中获取IL-16cDNA，插入PUC-18T载体并测序，构建并鉴定真核表达载体PcDNA3-IL-16，转染KarpasT淋巴瘤细胞，分析阳性克隆中IFNγ，IL-2，IL-4，IL-6，IL-13，TNFα和IL-16等细胞因子的表达状况。结果序列测定显示所克隆的人IL-16cDNA与基因库中所登录的序列完全一致，EcoRI和BamHI双酶切及PCR鉴定显示所构建的真核表达载体PcDNA3-IL-16完全正确，RT-PCR显示转染PcDNA3-IL-16的KarpasT淋巴瘤细胞高表达IL-16mRNA，同时，Th1类细胞因子IFNγ表达明显增高，Th2类细胞因子IL-4，IL-6，IL-13表达降低，MTT法显示KarpasT淋巴瘤细胞的增殖受到明显抑制。结论成功构建了真核表达载体PcDNA3-IL16，转染KarpasT淋巴瘤细胞后使其细胞因子类型由Th2型向Th0型逆转，且生长受到明显抑制，表明向肿瘤细胞转染人IL-16是一种有用的肿瘤生物治疗方法。

CONSTRUCTION OF HUMANIL-16 EUKARYOTIC EXPRESSION VECTOR AND ITS FUNCTION OF SWITCHING Th2 TYPE TUMOR CELLS

To construct human IL-16 eukaryotic expression vector,and analyse its function of switching Th2 type tumor cells. Methods: The human IL-16 cDNA was amplified by RT-PCR from peripheral blood mononuclear cells,and cloned into the PUC-18 T vector. Then the eukaryotic expression vector PcDNA3IL-16 containing IL-16 cDNA was constructed and identified. The Karpas T lymphoma cells were transfected with PcDNA-IL-16. Cytokines IFNγ, IL-2, IL-4, IL-6, IL-13,TNFα and IL-16 were detected by RT-PCR from the positive cell clones. Results:The sequence of cloned human IL-16 cDNA completely matched with that of human IL-16 cDNA in the gene bank. RT-PCR indicated the IL-16 mRNA was highly expressed in the Karpas cells after the recombinant vector PcDNA3-IL-16 was transfected into Karpas T lymphoma cells. The cells were switched from the expression of Th2 cytokines to Th0 type. The biological activity through MTT assay showed the proliferation of Karpas cells were sharply inhibited. Conclusion: The eukaryotic expression vector PcDNA31L-16 containing IL-16 cDNA was successfully constructed. The cytokine pattern of Karpas T lymphoma cells can be changed from Th2 type to Tho type by transfecting PcDNA-IL-16. These results suggest that is a useful method to transfect IL-16 gene into tumor cells for tumor biotherapy.