CAJ | 학술논문

目的培养胶质母细胞瘤干细胞,检测微小RNA-203(miR-203)在胶质母细胞瘤干细胞中的表达,并观察其对肿瘤干细胞增殖的影响.方法对胶质母细胞瘤组织行体外原代培养,以CD133为标志物,免疫磁珠法分选胶质母细胞瘤干细胞;免疫荧光染色检测分选所得细胞的CD133、巢蛋白(nestin)、神经胶质纤维酸性蛋白(GFAP)、微管相关蛋白2(MAP2)的表达,从而鉴定肿瘤干细胞.反转录-聚合酶链反应(RT-PCR)检测分选后CD133+细胞、CD133-细胞miR-203的表达;将miR-203模拟体和无意义寡核苷酸链(NC)分别转染胶质母细胞瘤干细胞作为miR-203、NC组,细胞计数试剂盒(CCK-8)法检测转染后24、48、72、96、120 h细胞生存率;流式细胞仪细胞检测转染后3d细胞凋亡率.结果培养的胶质母细胞瘤干细胞成球样生长,表达干细胞标志物CD133、Nestin,分化后细胞表达星形胶质细胞的标志物GFAP、神经元的标志物MAP2;分选后CD133+细胞中miR-203的表达低于CD133-细胞;转染miR-203后24、48、72、96、120 h组的细胞生存率均降低,差异有统计学意义(P＜0.05);流式细胞仪检测显示miR-203组的细胞凋亡率为(9.74±2.81)％,高于NC组的(3.95±0.91)％,差异有统计学意义(P＜0.05).结论上调miR-203可能成为针对胶质母细胞瘤干细胞的基因治疗策略.
목적 배양효질모세포류간세포,검측미소RNA-203(miR-203)재효질모세포류간세포중적표체,병관찰기대종류간세포증식적영향.방법 대효질모세포류조직행체외원대배양,이CD133위표지물,면역자주법분선효질모세포류간세포;면역형광염색검측분선소득세포적CD133、소단백(nestin)、신경효질섬유산성단백(GFAP)、미관상관단백2(MAP2)적표체,종이감정종류간세포.반전록-취합매련반응(RT-PCR)검측분선후CD133+세포、CD133-세포miR-203적표체;장miR-203모의체화무의의과핵감산련(NC)분별전염효질모세포류간세포작위miR-203、NC조,세포계수시제합(CCK-8)법검측전염후24、48、72、96、120 h세포생존솔;류식세포의세포검측전염후3d세포조망솔.결과 배양적효질모세포류간세포성구양생장,표체간세포표지물CD133、Nestin,분화후세포표체성형효질세포적표지물GFAP、신경원적표지물MAP2;분선후CD133+세포중miR-203적표체저우CD133-세포;전염miR-203후24、48、72、96、120 h조적세포생존솔균강저,차이유통계학의의(P＜0.05);류식세포의검측현시miR-203조적세포조망솔위(9.74±2.81)％,고우NC조적(3.95±0.91)％,차이유통계학의의(P＜0.05).결론 상조miR-203가능성위침대효질모세포류간세포적기인치료책략.
Objective To isolate glioblastoma multiforme stem cells (GBM-SCs) from glioblastoma multiforme (GBM) specimens and to investigate the expression of microRNA (miR)-203 in GBM stem cells and its impact on cell proliferation.Methods CD133 + cells were separated using magnetic cell sorting technique (MACS) after primary culture.Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze miR-203 expression in CD133 + cells.Lipofectamine RNAiMAX was used to transfect miR-203 mimic and scrambled control oligonucleotides into GBM-SCs.The proliferation ability of miR-203 treated GBM-SCs was determined by CCK-8 and the apoptosis ratio was detected by cell flow cytometry.Results GBM-SCs isolated from GBM specimens formed GBM spheres,expressed markers associated with neural stem cells,and possessed the capacity for self-renewal and multilineage differentiation.MiR-203 expression was down-regulated in CD133+ cells relative to CD133-cells.MiR-203 can repress GBM-SCs growth and induce apoptosis.Conclusion Reactivation of miR-203 expression suggests novel therapeutic strategies for GBM-SCs.