BRD4抑制剂对鼻咽癌CNE-2细胞增殖、凋亡和侵袭的影响

摘要

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、100μmol／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0μmol／L，凋亡率随浓度升高而增加；穿膜细胞数均少于0μmol／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0μmol／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。%Objective To explore the effects of bromodomain⁃containing protein 4 ( BRD4) inhibitor( GSK525762A) on pro⁃liferation, 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 in⁃hibitory 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 low⁃er than that of 0 mol/L group ( P<0. 05);Compared with the 0μmol/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.