中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2010年
2期
159-162
,共4页
靳峰%赵万巨%魏宇佳%张浩%张军臣%高超%郭强%邵彤%陈德勤%赵洪洋
靳峰%趙萬巨%魏宇佳%張浩%張軍臣%高超%郭彊%邵彤%陳德勤%趙洪洋
근봉%조만거%위우가%장호%장군신%고초%곽강%소동%진덕근%조홍양
神经胶质瘤%胶质瘤干细胞%凋亡%耐药
神經膠質瘤%膠質瘤榦細胞%凋亡%耐藥
신경효질류%효질류간세포%조망%내약
Glioma%Glioma stem cell%Apoptosis%Drug - resistance
目的 从人脑胶质瘤组织中分离、培养胶质瘤干细胞,并探讨其生物学特性及其抗凋亡和多重耐药基因的表达差异.方法 人脑胶质瘤组织经过原代细胞培养后,用无血清培养方法获得胶质瘤干细胞球,用10%的胎牛血清培养诱导分化,分化前后分别做nestin、tubulin-β、GFAP免疫细胞化学荧光染色,并观察胶质瘤干细胞形态学改变.同时,应用实时荧光定量PCR技术检测livin,livinot,livinβ,survivin,MRPI和MRP3 mRNA的表达.结果 有2例分离培养出干细胞球体,这些球体具有典型的干细胞特性,nestin染色为阳性;在无血清培养基中呈悬浮球样生长,能够自我更新和增殖,在有血清培养基中能够分化,tubulin-β、GFAP染色为阳性.胶质瘤干细胞球livin、livinα、livinβ、survivin和MRP-1 mRNA表达量比较胶质瘤组织表达量都有不同程度的升高,而MRP-3mRNA表达量降低.结论 胶质瘤干细胞抗凋亡和MRP-1基因表达量较胶质瘤表达量有不同程度的升高,提示胶质瘤干细胞比较其同源的胶质瘤细胞具有更强的耐药性,这可能是肿瘤耐药的机制.
目的 從人腦膠質瘤組織中分離、培養膠質瘤榦細胞,併探討其生物學特性及其抗凋亡和多重耐藥基因的錶達差異.方法 人腦膠質瘤組織經過原代細胞培養後,用無血清培養方法穫得膠質瘤榦細胞毬,用10%的胎牛血清培養誘導分化,分化前後分彆做nestin、tubulin-β、GFAP免疫細胞化學熒光染色,併觀察膠質瘤榦細胞形態學改變.同時,應用實時熒光定量PCR技術檢測livin,livinot,livinβ,survivin,MRPI和MRP3 mRNA的錶達.結果 有2例分離培養齣榦細胞毬體,這些毬體具有典型的榦細胞特性,nestin染色為暘性;在無血清培養基中呈懸浮毬樣生長,能夠自我更新和增殖,在有血清培養基中能夠分化,tubulin-β、GFAP染色為暘性.膠質瘤榦細胞毬livin、livinα、livinβ、survivin和MRP-1 mRNA錶達量比較膠質瘤組織錶達量都有不同程度的升高,而MRP-3mRNA錶達量降低.結論 膠質瘤榦細胞抗凋亡和MRP-1基因錶達量較膠質瘤錶達量有不同程度的升高,提示膠質瘤榦細胞比較其同源的膠質瘤細胞具有更彊的耐藥性,這可能是腫瘤耐藥的機製.
목적 종인뇌효질류조직중분리、배양효질류간세포,병탐토기생물학특성급기항조망화다중내약기인적표체차이.방법 인뇌효질류조직경과원대세포배양후,용무혈청배양방법획득효질류간세포구,용10%적태우혈청배양유도분화,분화전후분별주nestin、tubulin-β、GFAP면역세포화학형광염색,병관찰효질류간세포형태학개변.동시,응용실시형광정량PCR기술검측livin,livinot,livinβ,survivin,MRPI화MRP3 mRNA적표체.결과 유2례분리배양출간세포구체,저사구체구유전형적간세포특성,nestin염색위양성;재무혈청배양기중정현부구양생장,능구자아경신화증식,재유혈청배양기중능구분화,tubulin-β、GFAP염색위양성.효질류간세포구livin、livinα、livinβ、survivin화MRP-1 mRNA표체량비교효질류조직표체량도유불동정도적승고,이MRP-3mRNA표체량강저.결론 효질류간세포항조망화MRP-1기인표체량교효질류표체량유불동정도적승고,제시효질류간세포비교기동원적효질류세포구유경강적내약성,저가능시종류내약적궤제.
Objective To isolate and culture glioma stem cells from primary glioma cells and detect the expression of anti - apoptotic and multidrug resistance - associated protein (MRP) genes. Methods Glioma stem cells were isolated from human glioma cell by physics technique and nestin, β- tubulin and GFAP expression were examined by Immunofluoresence staining. Expression of livin, livinα, livinβ, survivin, MRPI and MRP3 were assayed by real - time quantitative RT-PCR. Results Glioma stem cells were induced successfully from two glioma specimens from one glioblastoma (No. 912110) and one grade 2 -3 astrocytoma subject (No. 916718). The glioma stem cells which were cultured in NSC medium grew in suspension and presented the features of stem cells: sphere - like shape, self - renewal and ability to differentiate. Nestin immunofluoresence staining of the stem cells presented positive, but the glioma did not show this feature. These cells could conditionally differentiate into tubulin -β~+ and GFAP~+ cells, which provided these cells in culture could produce neurons as well as glial cells. The mRNA expression of cancer stem cells on livin, livinα, survivin, MRP1 was significantly up - regulated than that of glioma primary cells. However, the mRNA expression of MRP3 was down - regulated. Conclusion Primary glioma cell contains cancer stem cells. The expression levels of anti - apeptotic and MRP1 genes are higher than that of glioma primary cells, which show glioma stem cells are the factor to maintain tumor growth and resist apoptosis and to pump the anti - tumor drugs out of cells, and this is what we need further research on.
