周冲,周菊英,王利利,俞志英,徐晓婷,秦颂兵,聂斌.miR-21反义寡核苷酸对SHG-44放射增敏作用的体外研究[J].中华放射医学与防护杂志,2010,30(6):701-704 |
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 |
投稿时间:2009-12-18 |
DOI: |
中文关键词: 胶质瘤 放射敏感性 miR-21 细胞周期 凋亡 |
英文关键词:Glioma Radiosensitivity miR-21 Cell Cycle Apoptosis |
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中文摘要: |
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 MeV X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。 |
英文摘要: |
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 oligonuleotides 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 significantely 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. |
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