CAJ | 학술논문

目的：探讨Notch1对人胶质瘤U251细胞干性和药物敏感性的影响。方法：用高表达Notch1胞内段（Notch1 intracellular domain， NICD1）和Notch1－shRNA慢病毒表达载体感染体外培养的人胶质瘤U251细胞， Western blot和免疫荧光染色法鉴定高表达NICD和Notch1沉默细胞。通过流式细胞术检测分析CD133＋细胞的比例、免疫荧光染色法检测nestin和GFAP的表达情况、检测肿瘤细胞球的形成率和SCID小鼠体内种植致瘤情况，分析Notch1对细胞干性的调节。并采用MTT法检测各组细胞对化疗药物替尼泊苷（VM－26）和卡莫司汀（BCNU）的敏感性。结果：NICD表达增加的瘤细胞干性表型增强，如CD133＋细胞的比例增加、nestin表达增强而GFAP表达减弱、肿瘤细胞球的形成率和SCID小鼠种植致瘤率增加，并伴有对VM－26和BCNU的敏感性降低。而Notch1基因表达下调的瘤细胞干性表型受到明显抑制，而对VM－26和BCNU的敏感性增高。结论：Notch1高表达可增加人胶质瘤U251细胞的干性，减弱U251细胞对化疗药物的敏感性。
목적：탐토Notch1대인효질류U251세포간성화약물민감성적영향。방법：용고표체Notch1포내단（Notch1 intracellular domain， NICD1）화Notch1－shRNA만병독표체재체감염체외배양적인효질류U251세포， Western blot화면역형광염색법감정고표체NICD화Notch1침묵세포。통과류식세포술검측분석CD133＋세포적비례、면역형광염색법검측nestin화GFAP적표체정황、검측종류세포구적형성솔화SCID소서체내충식치류정황，분석Notch1대세포간성적조절。병채용MTT법검측각조세포대화료약물체니박감（VM－26）화잡막사정（BCNU）적민감성。결과：NICD표체증가적류세포간성표형증강，여CD133＋세포적비례증가、nestin표체증강이GFAP표체감약、종류세포구적형성솔화SCID소서충식치류솔증가，병반유대VM－26화BCNU적민감성강저。이Notch1기인표체하조적류세포간성표형수도명현억제，이대VM－26화BCNU적민감성증고。결론：Notch1고표체가증가인효질류U251세포적간성，감약U251세포대화료약물적민감성。
AIM:To investigate whether Notch1 changes stemness and chemotherapeutic sensitivity in human glioma U251 cells.METHODS: The lentiviral vectors, which expressed Notch1-shRNA or Notch1 intracellular domain ( NICD) , were transfected into U251 cells .Western blot and immunofluorescence staining were applied to monitor the va-lidity of the cells, down-regulation of Notch1 expression or over-expression of NICD.The proportion of CD133 +cells was analyzed by flow cytometry.The expression of nestin and GFAP was identified by immunofluorescence staining.The forma-tion rate of tumor cell spheres and the implanted tumor growth in SCID mice were observed.MTT assay was performed to e-valuate the chemotherapeutic sensitivity to VM-26 and BCNU of the cells with different treatments.RESULTS:Stemness was significantly enhanced in the cells over-expressing NICD.For example, the proportion of CD133 +cells was increased, the expression of nestin was up-regulated, the expression of GFAP was down-regulated, and the formation rate of tumor cell spheres and implanted tumor growth were increased.The chemotherapeutic sensitivity to VM-26 and BCNU of the cells was decreased.In the cells with Notch1 gene down-regulation by RNAi, the stemness was inhibited and chemotherapeutic sensi-tivity was increased.CONCLUSION:Notch1, which leads to the change of stemness and chemotherapeutic sensitivity in human glioma U251 cells, is likely to be a potential molecular target for treatment of glioma.