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目的 探讨人骨形成蛋白 2 (rhBMP 2 )和透明质酸 (HA)对C6胶质细胞瘤体内侵袭的影响。方法 将携带增强型绿色荧光蛋白 (EGFP)基因的pEGFP N3质粒体外转染C6胶质瘤细胞 ,将C6阳性克隆以立体定向法植入SD大鼠脑实质内 ,建立大鼠胶质瘤移植模型。移植瘤区给予不同剂量的rhBMP 2和HA ,用病理检查、流式细胞计数、荧光显微镜及电子显微镜观察rhBMP 2和HA对胶质瘤侵袭的影响。结果 稳定转染EGFP基因的C6瘤细胞于体内、外均可发出绿色荧光 ,在荧光显微镜下易于区分肿瘤与非肿瘤区 ,并能检测到少量C6瘤细胞的远处侵袭性生长。 10 μl的 5 μg/mlrhBMP 2可明显抑制肿瘤体内侵袭 ;而 10 μl的 10 0 μg/mlHA则具有相反的作用。 结论 EGFP标记的C6瘤细胞移植于大鼠脑内 ,可建成稳定的脑胶质细胞瘤体内侵袭动物模型。rhBMP 2降低了C6瘤细胞的体内侵袭性 ,而HA则明显促进了其体内侵袭作用
Objective To investigate the effect of human bone morphogenetic protein 2 (rhBMP 2) and hyaluronic acid (HA) on the invasion of C6 glioma. Methods C6 glioma cells were transfected with pEGFP N3 plasmid carrying enhanced green fluorescent protein (EGFP) gene in vitro. C6 positive clones were implanted into brain parenchyma of SD rats stereotaxically to establish rat glioma transplantation model . Different dosages of rhBMP 2 and HA were given to the xenograft tumor area, and the effects of rhBMP 2 and HA on glioma invasion were observed by pathological examination, flow cytometry, fluorescence microscopy and electron microscopy. Results The C6 cells stably transfected with EGFP gene could both emit green fluorescence both in vitro and in vivo, and were easy to differentiate between tumor and non-tumor area under fluorescence microscope and to detect the invasive growth of small number of C6 tumor cells in distant place. 10 μl of 5 μg / ml rhBMP 2 significantly inhibited tumor invasion in vivo, whereas 10 μl of 10 μg / ml HA had the opposite effect. Conclusion The EGFP-labeled C6 tumor cells were transplanted into the brain of rats and established a stable invasive model of glioma. rhBMP 2 reduced the in vivo invasiveness of C6 tumor cells, whereas HA significantly promoted its in vivo invasion