| 张珠博,张仲健,魏超,翟贺争,阮书州,吴红英,李德冠,孟爱民,王小春.Cks1表达对人食管癌EC9706细胞辐射敏感性的影响[J].中华放射医学与防护杂志,2013,33(6):574-578 |
| Cks1表达对人食管癌EC9706细胞辐射敏感性的影响 |
| Effects of Cks1 expression on the radiosensitivity of esophageal cancer EC9706 cells |
| 投稿时间：2013-06-04 |
| DOI：10.3760/cma.j.issn.0254-5098.2013.06.002 |
| 中文关键词: 食管癌 Cks1 转染 辐射敏感性 |
| 英文关键词:Esophageal cancer Cks1 Transfection Radiosensitivity |
| 基金项目:国家自然科学基金面上项目（81272511）；天津市自然科学基金（11JCYBJC13700） |
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| 中文摘要: |
| 目的 通过多种转染方法改变人食管癌细胞EC9706中Cks1的表达，探究Cks1的表达对细胞辐射敏感性的影响。方法 通过3种转染方式构建人食管癌EC9706细胞模型，分别为Cks1正义转染组（以亲本组和空载组为对照）、Cks1 RNAi组（以亲本组和阴性对照组为对照）和Rescue组（转染了抗Cks1 RNAi的突变载体，以亲本组和RNAi组作为对照）。Western blot法检测转染后EC9706细胞中Cks1蛋白的表达量；6 Gy γ射线照射转染后的EC9706细胞，MTT法和克隆形成实验检测EC9706辐射敏感性的变化。结果 Western blot检测结果表明，转染成功改变了Cks1的表达量。正义转染实验中，相对于亲本组和空载组，Cks1正义转染组Cks1表达量提高（t=17.10、14.95，P<0.05）；RNA干扰实验中，相对于亲本组和阴性对照组，RNAi组表达量降低（t=3.85、6.37，P<0.05）；Rescue实验中，Rescue组较RNAi组表达量明显回升（t=7.72，P<0.05），接近亲本组的表达水平（P>0.05）。MTT实验结果表明，6 Gy γ射线照射后，Cks1正义转染组细胞的增殖能力显著高于亲本组（t=3.42、4.74、4.02，P<0.05）和空载组（t=3.39、4.44、4.37，P<0.05）。RNAi组细胞的增殖能力显著低于亲本组（t=7.33、8.27，P<0.05）和阴性对照组（t=6.98、7.34，P<0.05）。Rescue组细胞的增殖能力显著高于RNAi组（t=11.61、9.99，P<0.05），与亲本组相近（P>0.05）。克隆形成实验结果表明，Cks1正义转染组细胞克隆形成能力显著高于亲本组（t=13.95，P<0.05）和空载组（t=14.72，P<0.05），RNAi组细胞的克隆形成能力显著低于亲本组（t=20.14，P<0.05）和阴性对照组（t=10.01，P<0.05），Rescue组细胞的克隆形成能力显著高于RNAi组（t=10.57，P<0.05），与亲本组相近（P>0.05）。结论 Cks1的表达与食管癌细胞辐射敏感性密切相关，Cks1表达量越高，食管癌细胞辐射敏感性越低。 |
| 英文摘要: |
| Objective To research the effects of Cks1 expression on the radiosensitivity of esophageal cancer EC9706 cells after transfection of three kinds of methods. Methods Three kinds of transfections were carried on EC9706 cells, including Cks1 positive transfection group(compared to the parental group and vector group), Cks1 RNAi group (compared to the parental group and negative control group)and Rescue group (a mutant of Cks1 that was resistant to Cks1 siRNA, compared to the parental group and RNAi group). The expressions of Cks1 were measured by Western blot. EC9706 cells were exposed to γ-rays with a dose of 6 Gy. The changes of EC9706 radiosensitivity were measured by MTT and colony formation assay. Results The result of Western blot revealed that expressions of Cks1 were changed effectively by the three kinds of transfection. Compared with the parental group and vector group, the Cks1 expression of cells in positive transfection group was significantly improved(t=17.10, 14.95,P<0.05), while compared with the parental group and negative control group, that of cells in RNAi group was significantly reduced(t=3.85,6.37,P<0.05).However, the Cks1 expression of cells in Rescue group was obviously recovered(t=7.72,P<0.05), which was similar to that of cells in parental group(P>0.05). The result of MTT proved after exposure to γ-rays, cells in positive transfection group had a higher proliferation ability than those in the parental group(t=3.42, 4.74, 4.02,P<0.05)and the vector group(t=3.39, 4.44, 4.36,P<0.05).The cells in RNAi group had a lower proliferation ability than those in the parental group(t=7.33, 8.27,P<0.05)and the negative control group(t=6.98, 7.34,P<0.05). The cells in Rescue group had a higher proliferation ability than those in the RNAi group(t=11.61, 9.99,P<0.05),which was similar to those in parental group(P>0.05).Meanwhile, the result of colony formation assay revealed that cells with high expression level of Cks1 had a higher colony formation ability than those in the parental group(t=13.95,P<0.05) and the vector group (t=14.72,P<0.05).The cells in RNAi group had a lower colony formation ability than those in the parental group(t=20.14,P<0.05)and the negative control group (t=10.01,P<0.05). The cells in Rescue group had a higher colony formation ability than those in the RNAi group(t=10.57,P<0.05),which was similar to those in parental group(P>0.05). Conclusions The expression of Cks1 is closely related with the radiosensitivity of esophageal cancer cells EC9706. The radiosensitivity of esophageal cancer cells would be decreased with the down-regulation of Cks1 expression. |
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