IL-24对B16细胞生长抑制作用的体内外实验研究

目的:构建白细胞介素24(Interleukin24,IL24)真核表达质粒,研究其体内外表达对肿瘤细胞的生长抑制作用。方法:采用重组DNA技术构建IL24真核表达质粒pEGFPIL24。用脂质体法将重组质粒及空载体体外转染B16细胞,再经激光扫描共聚焦显微镜(LSM)观察其表达,用MTT法检测B16细胞的体外增殖能力,用流式细胞仪检测细胞周期。小鼠实体瘤模型研究基因转染对肿瘤的体内生长抑制作用。结果:pEGFPIL24转染B16细胞后,LSM可观察到IL24在细胞中的表达。转染IL24基因的B16细胞体外增殖能力明显受到抑制,细胞被阻滞在G2/M期。与对照组相比,IL24基因治疗组肿瘤在体内的生长受到明显抑制,P<0.05。结论:IL24基因转染的肿瘤细胞体外生长受抑。于荷瘤小鼠的瘤内注射pEGFPIL24可抑制肿瘤生长,提示IL24具有明显的体内外抗肿瘤作用。

Study of IL-24 suppressive effect on B16 cell line in vitro and in vivo

OBJECTIVE:A eukaryotic expressing plasmid of IL-24 was constructed , and its inhibitory effects on the growth of tumor cells were investigated in vitro and in vivo.METHODS: The eukaryotic expressing plasmid of IL-24 (pEGFP-IL-24) was constructed by DNA recombination technique. The recombination plasmid and empty vector were transfected into B16 cells with Lipofectamine 2000 reagent and the expression of IL-24 were determined by LSM; the proliferation of B16 cells was measured by MTT assay; and cell-cycle distribution of B16 cells were measured with Flow Cytometry. The inhibitory effect for solid tumor by IL-24 gene transfection in tumor carring mice was analyzed . RESULTS: The expression of IL-24 in B16 cells was determined by LSM after the transfection of pEGFP-IL-24. Comparing to the control group, the proliferation of B16 cells was inhibited by transfection with pEGFP-IL-24 and the G2/M phase of the transfected cells was also increased. Growth rate of tumor in tumor carring mice was significantly inhibited by IL-24 gene therapy as compared with the control group, P<0.05. CONCLUSIONS: Proliferation of B16 cells was inhibited by pEGFP-IL-24 transfection in vitro. The intratumor injection of pEGFP-IL-24 can remarkably inhibit the growth of solid tumor in mice.