中华放射医学与防护杂志

陈雪,高刚,阮健磊,等.外源表达miR-34a对H1299细胞辐射敏感性的影响[J].中华放射医学与防护杂志,2013,33(2):128-130.CHEN Xue,GAO Gang,RUAN Jian-lei,et al.Effects of exogenetic miR-34a on radiosensitivity of H1299 cells[J].Chin J Radiol Med Prot,2013,33(2):128-130

外源表达miR-34a对H1299细胞辐射敏感性的影响

Effects of exogenetic miR-34a on radiosensitivity of H1299 cells

投稿时间：2012-11-20

DOI：10.3760/cma.j.issn.0254-5098.2013.02.004

中文关键词: miR-34a H1299细胞辐射敏感性细胞凋亡细胞周期

英文关键词:miR-34a H1299 cells Radiosensitivity Apoptosis Cell cycle

基金项目:国家自然科学基金(31100606);卫生行业科研专项(201002009)

作者	单位	E-mail
陈雪	100088 北京,中国疾病预防控制中心辐射防护与核安全医学所辐射防护与核应急中国疾病预防控制中心重点实验室
高刚	100088 北京,中国疾病预防控制中心辐射防护与核安全医学所辐射防护与核应急中国疾病预防控制中心重点实验室
阮健磊	100088 北京,中国疾病预防控制中心辐射防护与核安全医学所辐射防护与核应急中国疾病预防控制中心重点实验室
刘建香	100088 北京,中国疾病预防控制中心辐射防护与核安全医学所辐射防护与核应急中国疾病预防控制中心重点实验室	jxliu@nirp.cn

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中文摘要:

目的研究miR-34a对H1299细胞辐射敏感性的影响。方法采用pre-miR-34a瞬时转染H1299细胞,接受不同剂量(0、2、4、6和8 Gy)γ射线照射后,利用CCK-8试剂盒检测细胞活性。流式细胞术检测细胞凋亡及细胞周期,real-time PCR检测miR-34a靶基因bcl-2、bax、CDK4、CDK6及cyclinD1的表达水平。结果不同剂量(0、2、4、6和8 Gy)γ射线照射后,与阴性对照转染组相比,pre-miR-34a转染组各剂量点细胞活力明显降低(t=-2.39、-3.12、-4.98、-4.03、-3.06,P<0.05),并与阴性对照转染组的差值呈剂量依赖性的增加。6 Gy γ射线照射后,与阴性对照转染组相比,pre-miR-34a转染组细胞凋亡率明显增加(t=7.06,P<0.05),细胞G₀/G₁期阻滞(t=3.94,P<0.05),S期细胞减少(t=6.23,P<0.05),bcl-2、CDK4及CDK6明显下调(t=3.39、12.88、6.21,P<0.05),cyclinD1有下降趋势,但差异无统计学意义,而bax明显上调(t=-4.35,P<0.05),是阴性对照转染组的1.94倍,bcl-2/bax比值下降。结论 miR-34促进H1299细胞凋亡,诱导细胞G₀/G₁期阻滞,抑制细胞活性,进而提高H1299细胞的辐射敏感性。

英文摘要:

Objective To investigate the influences of miR-34a on the radiosensitivity of H1299 cells. Methods CCK-8 kit was used to examine the viability of H1299 cells which were exposed to different doses (0, 2, 4, 6 and 8 Gy) of ⁶⁰Co γ-rays after transient transfection of pre-miR-34a. Apoptosis rate and cell cycle were measured with flow cytometry. The expression levels of miR-34a target genes, bcl-2, bax, CDK4, CDK6 and cyclinD1 were analyzed by real-time PCR. Results Compared to the control group of negative transfection, the cell viability in pre-miR-34a transfection group decreased significantly after irradiation at 0, 2, 4, 6, 8 Gy (t=-2.39, -3.12, -4.98, -4.03, -3.06, P<0.05) in a dose-dependent manner. After being irradiated with 6 Gy γ-rays, the apoptotic rate in pre-miR-34a transfection group was significantly increased (t=7.06, P<0.05) together with an accumulation of G₀/G₁ phase (t=3.94, P<0.05) and a reduction of S phase (t=6.23, P<0.05). The gene expression levels of bcl-2, CDK4 and CDK6 in pre-miR-34a transfection group were respectively decreased (t=3.39, 12.88, 6.21, P<0.05) of negative control. cyclinD1 was also decreased but no significance, while bax was increased to 1.94 times of negative control (t=-4.35, P<0.05) together with a decrease of bcl-2/bax. Conclusions miR-34a could promote cell apoptosis, induce G₀/G₁ phase accumulation, suppress cell activity, and in turn increase the radiosensitivity of H1299 cells.

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