CAJ | 학술논문

目的：探讨沉默ATP1A1基因对人U251胶质瘤干细胞增殖能力的影响。方法采用ATP1A1-shRNA慢病毒感染人U251胶质瘤干细胞，逆转录实时定量PCR（RT-qPCR）及蛋白质印迹法（Western blot）分别检测ATP1A1的mRNA和蛋白表达，流式细胞仪检测细胞周期和凋亡，M T T法检测细胞体外的增殖情况。结果通过RT-qPCR及Western blot 分析表明，感染后的细胞组ATP1A1的mRNA和蛋白表达均受到明显抑制，ATP1A1 mRNA抑制率为84．15％，蛋白表达抑制率为55．33％，细胞生长变慢，细胞凋亡比例显著增加，G1期细胞增多，而S期细胞减少，二者比较差异有统计学意义（ P＜0．05）。结论靶向ATP1A1基因干扰能够抑制人U251胶质瘤干细胞ATP1A1的表达，诱导细胞凋亡，调控细胞周期再分布，抑制细胞增殖。
목적：탐토침묵ATP1A1기인대인U251효질류간세포증식능력적영향。방법채용ATP1A1-shRNA만병독감염인U251효질류간세포，역전록실시정량PCR（RT-qPCR）급단백질인적법（Western blot）분별검측ATP1A1적mRNA화단백표체，류식세포의검측세포주기화조망，M T T법검측세포체외적증식정황。결과통과RT-qPCR급Western blot 분석표명，감염후적세포조ATP1A1적mRNA화단백표체균수도명현억제，ATP1A1 mRNA억제솔위84．15％，단백표체억제솔위55．33％，세포생장변만，세포조망비례현저증가，G1기세포증다，이S기세포감소，이자비교차이유통계학의의（ P＜0．05）。결론파향ATP1A1기인간우능구억제인U251효질류간세포ATP1A1적표체，유도세포조망，조공세포주기재분포，억제세포증식。
Objective To investigate the effects of ATP1A1 knockdown by RNA interference(RNAi) on proliferation of human U251 glioma stem cells .Methods The human U251 glioma stem cells were infected with lentivirus expressing ATP1A1-shRNA . The mRNA and protein expressions of ATP1A1 in U251 glioma stem cells were detected by RT-qPCR and Western blotting ,re-spectively .The cell cycle and apoptosis were evaluated by flow cytometry .The proliferation of U251 glioma stem cells was deter-mined by MTT assay .Results The expressions of ATP1A1 in U251 glioma stem cells transfected with ATP1A1-shRNA were in-hibited significantly at both mRNA and protein levels ,with an inhibitory rate of 84 .15% for ATP1A1 mRNA and of 55 .33% for ATP1A1 protein respectively .The proliferation of cells was inhibited ,the cell apoptotic rate was significantly increased and the cell cycle was arrested in G1 phase and S phase decreased significantly in ATP1A1-shRNA cells(P<0 .05) .Conclusion RNAi targe-ting ATP1A1 gene could down-regulates the ATP1A1 expression ,induces cell apoptosis ,regulates cell phase redistribution and in-hibits cell proliferation in U 251 glioma stem cells .