MicroRNA-181b提高U87细胞对VM-26的敏感性研究

miR-181b Enhances Sensitivity to Teniposide in Glioma Cell Line U87

背景与目的:MicroRNA(miRNA)参与肿瘤发生发展的诸多过程,并参与调节多种抗肿瘤药物的敏感性。本研究探讨恶性胶质瘤中miR-181b对VM-26(teniposide)化疗敏感性的影响。方法:以荧光定量PCR法检测miR-181b在高级别胶质瘤中的表达,并利用CCK-8细胞毒性实验检测高级别胶质瘤患者细胞对VM-26的化疗敏感性;并通过慢病毒感染构建稳定高表达miR-181b的U87/181b细胞及其对照组U87/nc,在荧光显微镜下观察其转染率及荧光定量PCR法检测其中miR-181b的表达;进而利用CCK-8细胞毒性实验检测U87/181b和U87/nc细胞对VM-26的敏感性,利用流式细胞仪检测VM-26作用72小时后U87/181b和U87/nc的凋亡情况。结果:在高级别胶质瘤中,miR-181b的表达与VM-26的敏感性呈正相关(r=-0.691,P﹤0.01),也就是miR-181b高表达肿瘤对VM-26的敏感性高。qPCR检测miR-181b在U87/181b(0.699±0.023)的表达显著高于U87/nc(0.019±0.001)(P﹤0.05)。CCK-8检测结果显示U87/181b[IC50:(1.25±0.12)μg/mL]对VM-26的敏感性显著高于U87/nc[IC50:(6.24±0.88)μg/mL](P﹤0.05)。经VM-26处理后U87/181b凋亡率(69.41±0.77)明显高于U87/nc(37.93±2.90)(P﹤0.05)。结论:在高级别胶质瘤高表达miR-181b的肿瘤对VM-26的敏感性高;在胶质瘤细胞U87中增加miR-181b表达可以提高对VM-26的敏感性。

BACKGROUND ＆ OBJECTIVE： miR-181b is low expressed in glioblastoma which is also related to chemotherapy resistance. In this study, we aimed to investigate whether miR-181b enhances sensitivity to the chemotherapeutic agent teniposide （VM-26） in gliomas. METHODS： The expression of miR-181b was measured by qRT-PCR in frozen human glioma specimens and CCK-8 was used to determine growth inhibiting effects of VM-26 on glioma specimens. U87 cells were transfeeted with miR-181b （U87/181b） or control miRNA （U87/nc） by using lentiviriral system. And then, the expression level of miR-181b was measured by qRT-PCR and cell viability was detected by CCK-8 assay. Apoptosis was detected by flow eytometry. RESULTS： In high grade human glioma with higher miR-181b expression revealed more sensitive to VM-26, with significantly positive correlation （r=-0.691, P 〈 0.01 ）. MiR-181b expression in U87/181b（0.699 ± 0.023）is significantly higher than in U87/ne（0.019±0.001 ） （P 〈 0.05）. CCK-8 assay showed that U87/181b （IC50： 1.25±0.12ug/ml）was more sensitive to VM-26 than U87/nc （ICS0： 6.24±0.88ug/ml）（P 〈 0.05）. U87/181b （69.41±0.77）had a higher apoptosis rate than U87/nc （37.93±2.90）（P 〈 0.05）. CONCLUSION： miR-181b expression level is positively correlated to VM-26 sensitivity in high grade gliomas. In glioma cell line U87, enhancing miR-181b expression may increase its sensitivity to VM-26.