BRD4抑制剂对鼻咽癌CNE-2细胞增殖、凋亡和侵袭的影响

目的 探讨溴结构域蛋白4（BRD4）抑制剂GSK525762A对鼻咽癌CNE-2细胞增殖、凋亡及侵袭的影响。方法 0、0.1、1、10、100 μmol／L GSK525762A 处理鼻咽癌CNE-2细胞 24、48、72和96 h后，采用四甲基偶氮唑盐（MTT）比色法检测细胞增殖抑制率变化，同时采用流式细胞术Annexin V-FITC／PI双染法检测不同浓度GSK525762A处理48、96 h后的CNE-2细胞凋亡情况，Transwell法检测不同浓度GSK525762A处理48、96 h后的CNE-2细胞侵袭能力，实时定量PCR检测不同浓度GSK525762A 处理48、96 h后凋亡相关基因的表达情况。结果 GSK525762A对CNE-2细胞增殖有抑制作用，增殖抑制率呈时间和浓度依赖性，差异有统计学意义（P＜0.05）；GSK525762A处理后的细胞早期、晚期及总凋亡率升高，均高于0 μmol／L，凋亡率随浓度升高而增加；穿膜细胞数均少于0 μmol／L，且随浓度升高而降低，以上差异均有统计学意义（P＜0.05）；与0 μmol／L比较，其余各浓度的Bcl-2 mRNA水平降低，Bax mRNA、Bak mRNA水平均升高，且各浓度间差异均有统计学意义（P＜0.05）；GSK525762A各浓度处理96 h的凋亡率、穿膜细胞数及凋亡相关基因mRNA均优于48 h（P＜0.05）。结论 BRD4抑制剂GSK525762A对鼻咽癌CNE-2细胞增殖有毒性作用，可诱导CNE-2细胞凋亡，恢复凋亡相关基因的表达，并降低细胞的侵袭能力。

Effects of BRD4 inhibitor on proliferation,apoptosis and invasion of nasopharyngeal carcinoma cell line CNE-2

Objective To explore the effects of bromodomain-containing protein 4 （BRD4） inhibitor（GSK525762A） on proliferation， apoptosis and invasion of nasopharyngeal carcinoma cell line CNE-2. Methods The nasopharyngeal carcinoma cell line CNE-2 was treated by GSK525762A （0， 0.1， 1， 10， 100 μmol／L） for 24， 48， 72 and 96 h. The methyl thiazolyl tetrazolium salt （MTT） was used to detect the proliferation inhibition rates at the above time points. Meanwhile， the apoptotic rates of CNE-2 cells at 48， 96 h after treatment with GSK525762A were measured with Annexin-FITC／PI double staining via flow cytometry. Transwell method was used to detect the invasion ability of CNE-2 cells at 48， 96 h after treatment with GSK525762A. The real time quantitative PCR was performed to detect expressions of apoptosis related genes at 48， 96 h after treatment with GSK525762A. Results There was inhibitory effect of GSK525762A on the proliferation of CNE-2 cells， and the inhibition rate was increased in a time- and concentration-dependent manner （P＜0.05）. After the treatment of GSK525762A， the early， late and total apoptotic rates increased with the increase of concentrations， higher than those of 0 mol／L group （P＜0.05）； the number of transmembrane cells of other concentrations were lower than that of 0 mol／L group （P＜0.05）； Compared with the 0 μmol／L， there were lower level of mRNA in Bcl-2 but higher levels of mRNA in Bax and Bak with the range （0.1-100 μmol／L） （P＜0.05）. Conclusion BRD4 inhibitor GSK525762A presented toxicity on the proliferation of CNE-2 cell， induced apoptosis and restored the expressions of apoptosis related genes.