长链非编码RNAHOTAIR对人胶质瘤细胞代谢和生长的影响  

Influence of long noncoding RNA HOTAIR on hmnan glioma cell metabolism and growth

作　　者：魏文金[1] 韩东风[1] 李海林[1] 张军霞[1] 张雅旋[1] 胡奇[1] 李文涛[1] 俞天赋 尤永平[1] 

机构地区：[1]南京医科大学第一附属医院神经外科,南京210029

出　　处：《中华神经外科杂志》2015年第9期942-947,共6页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金（81272792,81372709）;江苏省“科教兴卫工程”医学重点学科（RC2011051）

摘　　要：目的探讨长链非编码RNAHOX转录反义RNA （HOX transcript antisense RNA,HOTAIR）对人脑胶质瘤细胞代谢和生长的影响。方法采用荧光标记的小干扰HOTAIR（si—HOTAIR）转染胶质瘤U87和U251细胞株，通过实时定量荧光聚合酶链式反应（RT．PCR）检测HOTAIR在胶质瘤细胞中的表达水平；利用CCK8实验和克隆形成实验检测细胞的生长变化；采用海马生物公司糖酵解应激实验和线粒体应激实验试剂盒检测细胞有氧糖酵解和线粒体功能。通过Westernblot检测有氧糖酵解相关酶[包括葡萄糖-6-磷酸异构酶（GPI）、丙酮酸激酶M2（PKM2）及乳酸脱氢酶A（LDHA）]的表达水平。结果荧光显微镜下观察U87和U251细胞中si．HOTAIR转染效率均〉90％。与阴性对照（NC）组比较，si—HOTAIR组细胞中HOTAIR的表达均明显下调（P〈0．01）；si—HOTAIR组U87和U251胶质瘤细胞的增殖能力和克隆形成能力明显低于NC组（P〈0．01）；下调HOTAIR后，胶质瘤细胞有氧糖酵解能力减弱，表现为基础糖酵解水平降低（P〈0．05）、最大糖酵解能力减弱（P〈0．01）及糖酵解剩储备量下降（P〈0．01）；si—HOTAIR组GPI、PKM2和LDHA表达水平较NC组降低（P〈0．01）。下调HOTAIR后，胶质瘤细胞线粒体功能受损，与NC组比较，表现为基础呼吸水平下降（其中U87细胞P〈0．01）、线粒体偶联效率降低及剩余呼吸能力减弱（U87和U251组均P〈0．05）。结论下调HOTAIR表达可以使人胶质瘤细胞糖酵解能力和线粒体功能受损，同时影响胶质瘤细胞的生长能力。Objective To investigate the influence of long noneoding RNA HOX transcript antisense RNA （HOTAIR） on human glioma cell metabolism and growth. Methods The fluoreseently labeled small interfering （si-HOTAIR） was used to transfeet glioma U87 and U251 cell lines. The real-time polymerase chain reaction （RT-PCR） was used to detect the expression level of HOTAIR in glioma cells. CCK8 test and colony formation assay were used to detect the growth changes of the glioma cells. Seahorse BioSeienee XF glyeilysis stress test kit and XF mitoehondrial stress test kit were used to detect the cell aerobic glyeolysis and mitoehondrial function. Western blot was used to detect glyeolysis related enzymes, including the expression levels of glueose-6-phosphate isomerase （GPI） , Pyruvate kinase M2 （PKM2） , and lactate dehydrogenase A （LDHA）. Results The transfection efficiency of si-HOTAIR in U87and U251eells observed under a fluorescence microscope was more than 90%. The HOTAIR expression in the cells of the si-HOTAIR group was downregulated significantly compared with the negative control group （ P 〈 0. 01 ）. The proliferative ability and elonal formation ability of the U87and U251 glioma cells of the si-HOTAIR group were significantly lower that than those of the negative group （ P 〈 0. 01 ）. After downregulation of HOTAIR, the aerobic glyeolysis capacity of glioma cells were diminished, showing reduced based glyeolysis level （ P 〈 0. 05 ）, diminished maximum glyeolytie capacity （ P 〈 0. 01 ）, and decreased remaining amount of glyeolysis （P 〈 0. 01 ）. The expression levels of GPI, PKM2 and LDHA of the si-HOTAIR group were lower than those of the negative control group （P 〈 0.01 ）. After downregulation of HOTAIR, the mitochondrial function in glioma cells was impaired, showing decreased basal respiration level （U87 cells P 〈 0. 01 ）, decreased mitochondrial coupling efficiency, and diminished residual respiration ability （P 〈 0. 05）. Conclusions Dowrrregulatio

关 键 词：神经胶质瘤 线粒体 代谢 糖酵解 增殖 长链非编码RNA HOX转录反义RNA 

分 类 号：R739.4[医药卫生—肿瘤]