中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2013年
10期
726-731
,共6页
吴庭枫%陈金明%陈三送%陈桂林%韦永新%谢学顺%杜子威%周幽心
吳庭楓%陳金明%陳三送%陳桂林%韋永新%謝學順%杜子威%週幽心
오정풍%진금명%진삼송%진계림%위영신%사학순%두자위%주유심
神经胶质瘤%肿瘤干细胞%细胞,培养的%肿瘤移植%裸鼠
神經膠質瘤%腫瘤榦細胞%細胞,培養的%腫瘤移植%裸鼠
신경효질류%종류간세포%세포,배양적%종류이식%라서
Glioma%Tumor stem cell%Cells,cultured%Neoplasms transplantation%Nude mouse
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.
目的 研究SHG-44膠質瘤榦細胞毬的分子錶型、緻瘤能力及其移植瘤的病理學特徵.方法 應用神經榦細胞培養基(NSCM)培養SHG-44膠質瘤細胞株,穫取其膠質瘤榦細胞毬,純化培養後行細胞免疫(ICC)熒光染色檢測CD133、兔抗人巢蛋白(nestin)、A2B5、小鼠抗人波形蛋白(vimentin)、血管內皮細胞生長因子受體2(VEGFR-2)和IDH R132H的錶達；含血清培養基誘導其分化,ICC檢測CD133、nestin、vimentin、膠質纖維痠性蛋白(GFAP)、β-Ⅲ tubulin和半乳糖神經鞘氨醇酶(GalC)的錶達情況；建立SHG-44膠質瘤榦細胞毬原位移植瘤模型,免疫組織化學檢測CD133、nestin、VEGFR-2、GFAP、S-100和CD34的錶達；比較SHG-44細胞原位模型和SHG-44榦細胞毬移植瘤的病理學特徵.結果 應用神經榦細胞培養法能成功穫取SHG-44膠質瘤榦細胞毬.第10代SHG-44膠質瘤榦細胞毬和SHG-44膠質瘤細胞的CD133暘性細胞比例分彆為(71.63±5.92)％和(1.95±1.45)％.SHG-44膠質瘤榦細胞毬的nestin、vimentin、VEGFR-2和A2B5暘性細胞比例分彆為(84.06±7.58)％、(29.11 ±3.44)％、(64.44±3.69)％和(14.08 ±2.19)％.SHG-44膠質瘤榦細胞毬中可見IDH R132H突變的細胞群.含血清培養基誘導分化後,CD133、nestin、vimentin、GFAP、β-Ⅲ tubulin和GalC暘性細胞比例分彆為(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)％.原位移植瘤組織中GFAP、S-100和VEGFR-2蛋白錶達呈暘性,CD133和nestin蛋白錶達呈陰性.極少量特異抗人CD34暘性細胞圍繞成管狀.SHG-44膠質瘤榦細胞毬顱內原位移植瘤跼部浸潤能力高于SHG-44膠質瘤細胞顱內原位移植瘤.結論 神經榦細胞培養法可成功從SHG-44膠質瘤細胞株中穫取膠質瘤榦細胞,屬于CD133+ A2B5-亞群,高錶達VEGFR-2,具備自我更新及多嚮分化能力,可參與血管擬態的形成.
목적 연구SHG-44효질류간세포구적분자표형、치류능력급기이식류적병이학특정.방법 응용신경간세포배양기(NSCM)배양SHG-44효질류세포주,획취기효질류간세포구,순화배양후행세포면역(ICC)형광염색검측CD133、토항인소단백(nestin)、A2B5、소서항인파형단백(vimentin)、혈관내피세포생장인자수체2(VEGFR-2)화IDH R132H적표체；함혈청배양기유도기분화,ICC검측CD133、nestin、vimentin、효질섬유산성단백(GFAP)、β-Ⅲ tubulin화반유당신경초안순매(GalC)적표체정황；건립SHG-44효질류간세포구원위이식류모형,면역조직화학검측CD133、nestin、VEGFR-2、GFAP、S-100화CD34적표체；비교SHG-44세포원위모형화SHG-44간세포구이식류적병이학특정.결과 응용신경간세포배양법능성공획취SHG-44효질류간세포구.제10대SHG-44효질류간세포구화SHG-44효질류세포적CD133양성세포비례분별위(71.63±5.92)％화(1.95±1.45)％.SHG-44효질류간세포구적nestin、vimentin、VEGFR-2화A2B5양성세포비례분별위(84.06±7.58)％、(29.11 ±3.44)％、(64.44±3.69)％화(14.08 ±2.19)％.SHG-44효질류간세포구중가견IDH R132H돌변적세포군.함혈청배양기유도분화후,CD133、nestin、vimentin、GFAP、β-Ⅲ tubulin화GalC양성세포비례분별위(1.89±1.27)％、(6.67±2.75)％、(93.75±2.95)％、(91.33±4.75)％、(82.36±4.02)％화(8.92±3.19)％.원위이식류조직중GFAP、S-100화VEGFR-2단백표체정양성,CD133화nestin단백표체정음성.겁소량특이항인CD34양성세포위요성관상.SHG-44효질류간세포구로내원위이식류국부침윤능력고우SHG-44효질류세포로내원위이식류.결론 신경간세포배양법가성공종SHG-44효질류세포주중획취효질류간세포,속우CD133+ A2B5-아군,고표체VEGFR-2,구비자아경신급다향분화능력,가삼여혈관의태적형성.
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,A2B5,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-Ⅲ tubulin+ 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.Compared with the SHG-44 glioma cell-formed xenograft tumor,the SHG-44 glioma stem cell-formed xenograft tumor exhibited a higher local invasiveness.Conclusions SHG-44 glioma cell spheres are successfully collected after cultured under neural stem cell medium.They belong to the CD133 + A2B5-GSC subpopulation,highly expressing VEGFR-2,possess the ability of both self-renewal and multi-directional differentiation,and may participate in the formation of vasculogenic mimicry.
