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[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-q PCR)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 vs240±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。
[Objective] To observe the effect of JQ1 on the proliferation of cervical cancer HeLa cells and its possible mechanism. [Method] The effect of JQ1 on proliferation of HeLa cells was detected by CCK-8 colorimetric assay and divided into dimethylsulfoxide (DMSO) control group and JQ1 treatment group (final concentrations were 0.01, 0.1, 1 and 10 μmol / L) 24, 48, 72 and 120h respectively. The cells were divided into DMSO control group and 10μmol / L JQ1 treatment group for follow-up experiments. After 12 days of cell culture, the number of cell clone formation was calculated. The apoptosis of cells was detected by flow cytometry after 72 hours culture. Real- RT-q PCR) and Western blotting were used to detect the expression of S-phase kinase-related protein 2 (SKP2) and p27. [Results] JQ1 inhibited the proliferation of HeLa cells in a time-and dose-dependent manner. After treated with 10μmol / L JQ1 for 72h, the cell viability was reduced by half. Compared with DMSO control group, JQ1 at 10μmol / L significantly inhibited the colony formation of HeLa cells The number of cell clones formed was 3 ± 2 vs 240 ± 10, P <0.001), and the apoptosis of HeLa cells was also induced (the percentage of apoptosis was 12.80 ± 0.88 vs 2.90 ± 0.27, P <0.01). Compared with DMSO control group, JQ1 could inhibit the expression of SKP2 mRNA and protein (SKP2 mRNA: 0.43 ± 0.02 vs 1.00 ± 0.03, P <0.01) and up-regulated the expression of p27 mRNA and protein (p27 mRNA: 2.60 ± 0.13 vs 1.00 ± 0.11, P <0.01). [Conclusion] JQ1 can inhibit the proliferation of HeLa cells by inducing apoptosis. The possible mechanism is that JQ1 inhibits the expression of SKP2 and up-regulates the expression of p27.