SHG-44胶质瘤干细胞球的表型特点及其移植瘤病理特征  

Phenotype of SHG-44 glioma stem cell spheres and pathological characteristics of their xenograft tumors

作　　者：吴庭枫[1] 陈金明[1] 陈三送[1] 陈桂林[1] 韦永新[1] 谢学顺[1] 杜子威[1] 周幽心[1] 

机构地区：[1]苏州大学附属第一医院神经外科,215006

出　　处：《中华肿瘤杂志》2013年第10期726-731,共6页Chinese Journal of Oncology

基　　金：江苏省卫生厅重点科研课题（K201106）

摘　　要：目的 研究SHG-44胶质瘤干细胞球的分子表型、致瘤能力及其移植瘤的病理学特征.方法 应用神经干细胞培养基（NSCM）培养SHG-44胶质瘤细胞株,获取其胶质瘤干细胞球,纯化培养后行细胞免疫（ICC）荧光染色检测CD133、兔抗人巢蛋白（nestin）、A2B5、小鼠抗人波形蛋白（vimentin）、血管内皮细胞生长因子受体2（VEGFR-2）和IDH R132H的表达；含血清培养基诱导其分化,ICC检测CD133、nestin、vimentin、胶质纤维酸性蛋白（GFAP）、β-Ⅲ tubulin和半乳糖神经鞘氨醇酶（GalC）的表达情况；建立SHG-44胶质瘤干细胞球原位移植瘤模型,免疫组织化学检测CD133、nestin、VEGFR-2、GFAP、S-100和CD34的表达；比较SHG-44细胞原位模型和SHG-44干细胞球移植瘤的病理学特征.结果 应用神经干细胞培养法能成功获取SHG-44胶质瘤干细胞球.第10代SHG-44胶质瘤干细胞球和SHG-44胶质瘤细胞的CD133阳性细胞比例分别为（71.63±5.92）％和（1.95±1.45）％.SHG-44胶质瘤干细胞球的nestin、vimentin、VEGFR-2和A2B5阳性细胞比例分别为（84.06±7.58）％、（29.11 ±3.44）％、（64.44±3.69）％和（14.08 ±2.19）％.SHG-44胶质瘤干细胞球中可见IDH R132H突变的细胞群.含血清培养基诱导分化后,CD133、nestin、vimentin、GFAP、β-Ⅲ tubulin和GalC阳性细胞比例分别为（1.89±1.27）％、（6.67±2.75）％、（93.75±2.95）％、（91.33±4.75）％、（82.36±4.02）％和（8.92±3.19）％.原位移植瘤组织中GFAP、S-100和VEGFR-2蛋白表达呈阳性,CD133和nestin蛋白表达呈阴性.极少量特异抗人CD34阳性细胞围绕成管状.SHG-44胶质瘤干细胞球颅内原位移植瘤局部浸润能力高于SHG-44胶质瘤细胞颅内原位移植瘤.结论 神经干细胞培养法可成功从SHG-44胶质瘤细胞株中获取胶质瘤干细胞,属于CD133＋ A2B5-亚群,高表达VEGFR-2,具备自我更新及多向分化能力,可参与血管拟态的形成.Objective To study the phenotype and tumorigenicity of SHG-44 glioma stem cell spheres and the pathological characteristics of their xenograft tumors. Methods SHG-44 glioma cells were cultured under neural stem cell medium and glioma stem cell spheres were collected. Immunocytochemistry was used to dectet the expression of CD133, nestin, A2BS, vimentin, VEGFR-2 and IDH R132H. Cell spheres were induced using serum-containing medium, and the expression of CD133, nestin, vimentin, GFAP, β-Ⅲ tubulin and GalC in the cell spheres were detected. The expression of CD133, nestin, VEGFR-2, GFAP, S-100 and CD34 in the intracranial xenograft tumor tissues was detected using immunohistochemistry. The pathological characteristics of orthotopic xenograft tumors generated from the SHG-44 glioma cells and SHG-44 glioma stem cell spheres were compared. Results SHG-44 glioma stem cell spheres were collected successfully after cultured under neural stem cell medium. The ratio of CD133 ＋ cells in the passage 10 SHG-44 glioma stem cell spheres was （71.63 ±5.92）%, significantly higher than that in the SHG-44 glioma cells [ （ 1.95 ±1.45 ） % ]. Immunocytochemistry showed that in the SHG-44 glioma cell spheres, the ratio of nestin ＋ cells was （84.06 ± 7.58） %, vimentin＋ cells （29.11 ± 3.44） %,VEGFR 2＋ cells （64.44±3.69）%, and A2B5＋ cells （14.08 ±2.19）%. A subpopulation of cells with mutation of IDH R132H was detected harboring in the SHG-44 glioma cell spheres. After induction of differentiation with serum-containing medium, the ratio of CD133 ＋ cells was （1.89 ± 1.27 ）% , nestin ＋cells （6.67±2.75）%, vimentin＋cells （93.75 ±2.95）%, GFAP＋ cells （91.33±4.75）%, β-Ⅲtubulin ＋ cells （ 82. 36 ± 4.02 ） %, and GalC ＋ cells （ 8.92 ± 3. 19 ） %. Immunohistochemistry showed positive expression of GFAP, S-100, VEGFR-2, and negative of CD133 and nestin in the orthotopic xenograft tumors. A very small amount of human-specific CD34 cells formed a tubular structure. Comp

关 键 词：神经胶质瘤 肿瘤干细胞 细胞 培养的 肿瘤移植 裸鼠 

分 类 号：R739.4[医药卫生—肿瘤]