BRD4抑制剂JQ1对宫颈癌HeLa细胞增殖、凋亡和侵袭作用的研究

Study on the Effect of BRD4 Inhibitor JQ1 on the Proliferation,Apoptosis and Invasion of Cervical Cancer Cell Line HeLa

目的:探讨溴结构域蛋白4(BRD4)抑制剂JQ1对宫颈癌HeLa细胞的增殖、凋亡及侵袭的影响,以及JQ1的抗肿瘤作用机制。方法:采用0、0.01、0.1、1μmol/L的JQ1处理宫颈癌HeLa细胞,分别于处理后24、48、72、120h采用CCK8检测细胞增殖情况,同时采用流式细胞术Annexin V-FITC/PI双染法检测0.5μmol/L和1μmol/L JQ1处理48h后的HeLa细胞凋亡情况,采用划痕试验检测细胞侵袭能力,实时定量PCR检测不同浓度JQ1处理48h后c-Myc及Bcl-2的表达情况。结果:与对照组相比,接触CCK-8试剂48h后,接受JQ1处理的HeLa细胞增殖受到明显抑制,具有剂量—时间依赖性(P<0.05)。对照组HeLa细胞凋亡率为20.38%,而JQ1(0.5μmol/L)组和JQ1(1μmol/L)组凋亡率分别为26.90%和40.33%,组间比较差异具有统计学意义(P<0.05)。细胞划痕试验提示JQ1对HeLa细胞的侵袭能力产生显著的抑制作用。经JQ1处理后的HeLa细胞c-Myc及Bcl-2表达量下降,并且随着JQ1浓度的增加mRNA表达量逐渐下降。结论:JQ1可以显著抑制HeLa细胞的增殖、侵袭能力,诱导HeLa细胞凋亡并抑制c-Myc及Bcl-2的表达,是一种对宫颈癌具有潜在抗肿瘤能力的药物。

Objective:To investigate the effect of bromodomain protein 4(BRD4) inhibitor JQ1 on the proliferation, apoptosis and invasion of cervical cancer cell line HeLa, and to explore the anti-tumor mechanism of JQ1.Methods:Cervical cancer cells were treated with JQ1(0,0.01,0.1,1μmol/L).CCK8 assay was used to detect cell proliferation at 24, 48, 72, and 120 hours after treatment. Flow cytometry Annexin V-FITC/PI double staining method was used to detect the apoptosis of HeLa cells treated with 0.5μmol/L and 1μmol/L JQ1 for 48 hours. The scratch test was used to detect the cell invasion ability, and the quantitative real-time PCR(qRT-PCR) was used to detect the expression of c-Myc and Bcl-2 after 48 hours of HeLa cells treated with different concentrations of JQ1.Results:The proliferation of HeLa cell treated with JQ1 was inhibited obviously after exposed to CCK-8 for 48 hours with dosage-time dependency comparing to those in control. The apoptosis rate of HeLa cells in the control group was 20.38%, while that in the JQ1(0.5μmol/L) group and JQ1(1μmol/L) group were 26.90% and 40.33%, respectively, and the difference was statistically significant(P<0.05). The cell scratch test indicated that JQ1 had a significant inhibitory effect on the invasion ability of HeLa cells. After JQ1 treatment, the expression of c-Myc and Bcl-2 in HeLa cells decreased with dosage dependency.Conclusion:JQ1 can significantly inhibit the proliferation and invasion of HeLa cells, induce the apoptosis and inhibit the expression of c-Myc and Bcl-2, which is a potential anti-tumor drug for cervical cancer.