王丽君,肖仲清,姚 亮.JQ1下调SKP2和上调p27诱导HeLa细胞凋亡的研究[J].肿瘤学杂志,2016,22(7):533-537.
JQ1下调SKP2和上调p27诱导HeLa细胞凋亡的研究
Study on JQ1 Inducing Apoptosis by Down-regulation of SKP2 and Up-regulation of p27 in HeLa Cells
投稿时间:2015-09-19  
DOI:10.11735/j.issn.1671-170X.2016.07.B002
中文关键词:  JQ1  HeLa细胞  细胞凋亡  S期激酶相关蛋白2  p27
英文关键词:JQ1  HeLa cells  cell apoptosis  S-phase kinase-associated protein 2(SKP2)  p27
基金项目:
作者单位
王丽君 江西省妇幼保健院 
肖仲清 江西省妇幼保健院 
姚 亮 江西省妇幼保健院 
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中文摘要:
      摘 要:[目的] 观察JQ1对宫颈癌HeLa细胞增殖的影响及可能的作用机制。[方法] 采用CCK-8比色法检测JQ1对HeLa细胞增殖的影响,分为二甲亚砜(DMSO)对照组和JQ1处理组(终浓度分别为0.01、0.1、1和10μmol/L),分别处理24、48、72和120h。将细胞分为DMSO对照组和10μmol/L JQ1处理组进行后续实验,细胞培养12d后计算细胞克隆形成数量;细胞培养72h后采用流式细胞仪检测细胞凋亡的变化,采用实时荧光定量PCR(RT-qPCR)和蛋白印迹法检测S期激酶相关蛋白2(SKP2)和p27的表达变化。[结果] JQ1抑制HeLa细胞增殖,且作用呈剂量与时间依赖性,10μmol/L JQ1处理细胞72h后细胞存活数量减半;与DMSO对照组相比,10μmol/L JQ1显著抑制HeLa细胞克隆形成(细胞克隆形成数量:3±2 vs 240±10,P<0.001);且能诱导HeLa细胞凋亡(细胞凋亡百分比:12.80±0.88 vs 2.90±0.27,P< 0.01);与DMSO对照组相比,JQ1能抑制细胞SKP2 mRNA 和蛋白表达(其中SKP2 mRNA表达倍数:0.43±0.02 vs 1.00±0.03,P<0.01),并上调p27 mRNA 和蛋白表达(p27 mRNA表达倍数:2.60±0.13 vs 1.00±0.11,P<0.01)。[结论] JQ1通过诱导细胞凋亡来抑制HeLa细胞的增殖,其可能的机制是抑制SKP2表达和上调p27表达。
英文摘要:
      Abstract:[Objective] To investigate the effect of JQ1 on proliferatoin of HeLa cells and its potential mechanism.[Methods] HeLa cells were exposed to dimethyl sulfoxide (DMSO,control group) or different concentrations of JQ1(0.01,0.1,1 and 10μmol/L,JQ1 group) in the culture medium for 24,48,72 and 120h respectively,and cell proliferation analysis was performed using CCK-8 method. HeLa cells were exposed to DMSO or 10μmol/L JQ1 in the culture medium for 12d,and the number of colony formation was assayed by counting. HeLa cells were exposed to DMSO or 10μmol/L JQ1 in the culture medium for 72h,and the apoptosis was assayed by flow cytometry,and the expression of S-phase kinase-associated protein 2(SKP2) and p27 were detected by real-time quantitative PCR (RT-qPCR) and Western blot methods. [Results] The proliferation of HeLa cells was inhibited by JQ1 in a time- and dose-dependent manner,a fifty percent inhibition of growth was seen after treated with 10μmol/L concentration of JQ1 for 72h. Compared with DMSO control group,the colony formation was inhibited significantly in 10μmol/L JQ1-treated group (the number of colony formation:3±2 vs 240±10,P<0.001);and cell apoptosis was induced signiticantly by 10μmol/L JQ1 (the percentage of apoptosis:12.80±0.88 vs 2.90±0.27,P<0.01). Compared with DMSO control group,the expression of SKP2 was significantly decreased and the expression of p27 was significantly increased in JQ1-treated group (among them SKP2 mRNA expression folds:0.43±0.02 vs 1.00±0.03;p27 mRNA expression folds:2.60±0.13 vs 1.00±0.11,both with P<0.01). [Conclusion] JQ1 inhibits cell proliferation of HeLa cells via inducing apoptosis,which might depend on down-regulation of SKP2 and up-regulation of p27.
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