人骨形成蛋白-2和透明质酸对脑胶质细胞瘤侵袭性的影响

Effects of recombinant human bone morphogenic protein-2 and hyaluronic acid on invasion of brain glioma in vivo

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 。

Objective To investigate the effects of recombinant human bone morphogenic protein 2(rhBMP 2)and hyaluronic acid (HA) on the invasion and metastasis of rat brain glioma in vivo. Methods C6 rat glioma cells were transfected with the plasmid vector named pEGFP N3 which contained an enhanced green fluorescent protein (EGFP) gene. The stable cell clones which expressed EGFP protein were stereotactically injected into the brain parenchyma of SD rats in order to establish a xenotransplanted tumor model. After rhBMP 2 and HA were administered in the xenotransplanted tumor model at different dosages, their effects on glioma invasion and metastasis were observed by pathology, flow cytometry fluoresclnec, and electronmicvoscopy. Results EGFP transfected C6 glioma cells gave off green fluorescence in vivo and in vitro. Area with tumor and area without tumor could be easily distinguished by fluorescence microscopy. Even distant micrometastasis and invasion at single cell level could be detected. 10 μl rhBMP 2 of the concentration of 5 μg/ml inhibited C6 tumor invasion, while 10 μl hyaluronic acid of the concentration of 100 μg/ml evidently promoted the tumor invasion. Conclusion EGFP transfected C6 cells can be transplanted into rats′ brain so as to establish an excellent experimental animal model in the research on invasion and metastasis of brain glioma in vivo. RhBMP 2 inhibits tumor invasion, while HA evidently promotes its invasion.