| 刘艳杰,顾浩,乔玉环,史惠蓉,张梦真,边爱平,纪妹,郭红军,常青,郭瑞霞.沉默细胞周期检测点激酶1(Chk1)对宫颈癌HeLa细胞放射敏感性的影响[J].中华放射医学与防护杂志,2017,37(2):102-106 |
| 沉默细胞周期检测点激酶1(Chk1)对宫颈癌HeLa细胞放射敏感性的影响 |
| Effect of silencing Chk1 expression on radiosensitivity of human cervical carcinoma HeLa cells |
| 投稿时间：2016-07-03 |
| DOI：10.3760/cma.j.issn.0254-5098.2017.02.004 |
| 中文关键词: RNA干扰 Chk1 宫颈癌 放射敏感性 |
| 英文关键词:RNA interference Checkpoint kinase 1 Cervical cancer Radiosensitivity |
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| 中文摘要: |
| 目的 探讨siRNA沉默细胞周期检测点激酶1（Chk1）基因对宫颈癌HeLa细胞放射敏感性的影响。方法 合成Chk1基因的小分子干扰RNA（si-Chk1），将si-NC或者si-Chk1分别转染到宫颈癌HeLa细胞中，0、2、4、6、8、10 Gy剂量的X射线照射细胞，RT-qPCR和Western blot检测转染后Chk1的mRNA水平和蛋白水平的表达，MTT方法检测细胞增殖能力，流式细胞仪检测细胞凋亡，克隆形成实验检测转染后HeLa细胞放射敏感性，Western blot检测转染后P21蛋白和抗凋亡基因Bcl-2蛋白的表达水平。结果 si-Chk1转染到HeLa细胞后，HeLa细胞Chk1基因的mRNA和蛋白表达水平均显著下降（t=2.12~5.86，P<0.05），沉默Chk1的表达抑制了宫颈癌HeLa细胞的增殖（t=3.15~6.36，P<0.05），同时降低宫颈癌HeLa细胞克隆形成能力（t=1.58~6.36，P<0.05），上调P21（t=4.35，P<0.05）、下调Bcl-2蛋白（t=2.37，P<0.05）的表达水平。结论 siRNA沉默Chk1表达能够增强HeLa细胞放射敏感性。 |
| 英文摘要: |
| Objective To study the effect of checkpoint kinase 1(Chk1) knock-down on radiosensitivity of human cervical carcinoma HeLa cells.Methods Chk1 was knock-down by using a specific siRNA of Chk1 gene. After transfection with si-CHK1 or its negative control si-NC, HeLa cells were irradiated with 0, 2, 4, 6, 8 and 10 Gy of X-rays. The expressions of Chk1 mRNA and protein in irradiated HeLa cells were determined by RT-qPCR and Western blot, respectively. Cell proliferation was measured by MTT assay, apoptosis was measured by flow cytometry assay, radiosensitivity was determined by colony formation, and the protein expressions of P21 and Bcl-2 were determined by Western blot. Results After transfection of si-Chk1, both mRNA and protein levels of Chk1 in HeLa cells were decreased(t=2.12-5.86, P<0.05). Chk1 knock-down decreased cell proliferation(t=3.15-6.57, P<0.05), reduced colony formation ability(t=1.58-6.36, P<0.05), increased the protein expression level of P21(t=4.35, P<0.05)and decreased the protein level of Bcl-2(t=2.37, P<0.05)in the irradiated HeLa cells in comparison with nonirradiated control.Conclusions siRNA-mediated knock-down of Checkpoint kinase 1 enhances the radiosensitivity of HeLa cells. |
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