miR-21反义寡核苷酸对SHG-44放射增敏作用的体外研究  

Study of antisense oligonucleotide miR-21 on radiosensitivity of SHG-44 in vitro

作　　者：周冲[1] 周菊英[1] 王利利[1] 俞志英[1] 徐晓婷[1] 秦颂兵[1] 聂斌[1] 

机构地区：[1]苏州大学附属第一医院放疗科,215006

出　　处：《中华放射医学与防护杂志》2010年第6期701-704,共4页Chinese Journal of Radiological Medicine and Protection

摘　　要：目的 探讨抑制miR-21表达对SHG-44人脑胶质瘤细胞系放射敏感性的影响及其机制.方法 脂质体介导采用瞬间转染法转染反义miR-21寡核苷酸（AS-miR-21）及阴性对照寡核苷酸于胶质瘤SHG-44细胞中.设空白对照组、阴性对照转染组及AS-miR-21转染组,分别给予6 MeVX射线单次照射0、1、2、4、6和8 Gy,采用成克隆实验计算放射增敏比,并绘制细胞存活曲线.流式细胞仪检测上述3组及联合单次6 Gy照射后细胞凋亡率、细胞周期变化.结果 转染AS-miR-21组的D0、Dq较空白对照组及阴性转染组降低,放射增敏比SERD0、SERDq分别为1.32和2.10.细胞周期分析示,转染组较空白对照及阴性对照组的G0/G1期比例增高,S期比例降低（t=8.18、-4.52,P＜0.05）.凋亡分析示,单纯照射组、AS-miR-21转染组及照射联合AS-miR-21转染组的早期凋亡率均高于对照组（t=20.14、11.11、50.07,P＜0.05）.结论 AS-miR-21可增加胶质瘤SHG-44细胞系放射敏感性,其机制可能与促进细胞凋亡及周期再分布有关.Objective To investigate the radiosensitizing effect of knock-down the expression of miR-21 on human SHG-44 glioma cells and explore the possible mechanism. Methods Antisense oligonuleotidas of miR-21, mediated by LipofectamineTM 2000, were transfected to SHG-44 cells. Three groups were： blank control group （ mock group）, negative control and antisense transfected group （ AS-miR-21 gorup）. Cells of each group were irradiated with 6 MeV X-rays at the doses of 0,1,2,4,6 and 8 Gy.Dose-suvivial curve was established by colony-forming assay. The influence of AS-miR-21 on cell cycle and cell apoptosis was analyzed by flow cytometry assay after 6 Gy irradiation. Results The value of D0 and Dq of AS-miR-21 group declined obviously compared with the mock group and negative control group. Flow cytometric analysis showed that cell cycle distribution changed（ G0/G1 phase arrest, S phase decreased）after transfected with AS-miR-21 （t = 8.18, -4.52,P ＜ 0.05 ）. The sensitization enhancement ratios of D0 and Dq were 1.32 and 2.10 respectively. Apoptosis assay showed the early apoptosis rate was signiflcantely increased in AS-miR-21 、irradition alone and combined group than mock control group（ t = 20.14,11.11,50.07, P ＜ 0.05）. Conclusions AS-miR-21 can enhance the radiosensitivity of human glioma cells SHG-44 by promoting cell apoptosis and faciliating cell cycle redistribution.

关 键 词：胶质瘤 放射敏感性 MIR-21 细胞周期 凋亡 

分 类 号：R739.41[医药卫生—肿瘤]