临床肿瘤学杂志
臨床腫瘤學雜誌
림상종류학잡지
CHINESE CLINICAL ONCOLOGY
2015年
6期
501-505
,共5页
刘忠%杨飞%程吉%田天捷
劉忠%楊飛%程吉%田天捷
류충%양비%정길%전천첩
溴结构域蛋白4%鼻咽癌%GSK525762A%增殖%凋亡
溴結構域蛋白4%鼻嚥癌%GSK525762A%增殖%凋亡
추결구역단백4%비인암%GSK525762A%증식%조망
Bromodomain-containing protein 4(BRD4)%Nasopharyngeal carcinoma%GSK525762A%Proliferation%Apoptosis
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。
目的：探討溴結構域蛋白4（ BRD4）抑製劑GSK525762A對鼻嚥癌CNE?2細胞增殖、凋亡及侵襲的影響。方法0、0．1、1、10、100μmol／L GSK525762A處理鼻嚥癌CNE?2細胞24、48、72和96 h後，採用四甲基偶氮唑鹽（ MTT）比色法檢測細胞增殖抑製率變化，同時採用流式細胞術Annexin V?FITC／PI雙染法檢測不同濃度GSK525762A處理48、96 h後的CNE?2細胞凋亡情況，Transwell法檢測不同濃度GSK525762A處理48、96 h後的CNE?2細胞侵襲能力，實時定量PCR檢測不同濃度GSK525762A 處理48、96 h後凋亡相關基因的錶達情況。結果 GSK525762A對CNE?2細胞增殖有抑製作用，增殖抑製率呈時間和濃度依賴性，差異有統計學意義（ P＜0．05）；GSK525762A處理後的細胞早期、晚期及總凋亡率升高，均高于0μmol／L，凋亡率隨濃度升高而增加；穿膜細胞數均少于0μmol／L，且隨濃度升高而降低，以上差異均有統計學意義（ P＜0．05）；與0μmol／L比較，其餘各濃度的Bcl?2 mRNA水平降低，Bax mRNA、Bak mRNA水平均升高，且各濃度間差異均有統計學意義（ P＜0．05）；GSK525762A各濃度處理96 h的凋亡率、穿膜細胞數及凋亡相關基因mRNA均優于48 h（ P＜0．05）。結論 BRD4抑製劑GSK525762A對鼻嚥癌CNE?2細胞增殖有毒性作用，可誘導CNE?2細胞凋亡，恢複凋亡相關基因的錶達，併降低細胞的侵襲能力。
목적：탐토추결구역단백4（ BRD4）억제제GSK525762A대비인암CNE?2세포증식、조망급침습적영향。방법0、0．1、1、10、100μmol／L GSK525762A처리비인암CNE?2세포24、48、72화96 h후，채용사갑기우담서염（ MTT）비색법검측세포증식억제솔변화，동시채용류식세포술Annexin V?FITC／PI쌍염법검측불동농도GSK525762A처리48、96 h후적CNE?2세포조망정황，Transwell법검측불동농도GSK525762A처리48、96 h후적CNE?2세포침습능력，실시정량PCR검측불동농도GSK525762A 처리48、96 h후조망상관기인적표체정황。결과 GSK525762A대CNE?2세포증식유억제작용，증식억제솔정시간화농도의뢰성，차이유통계학의의（ P＜0．05）；GSK525762A처리후적세포조기、만기급총조망솔승고，균고우0μmol／L，조망솔수농도승고이증가；천막세포수균소우0μmol／L，차수농도승고이강저，이상차이균유통계학의의（ P＜0．05）；여0μmol／L비교，기여각농도적Bcl?2 mRNA수평강저，Bax mRNA、Bak mRNA수평균승고，차각농도간차이균유통계학의의（ P＜0．05）；GSK525762A각농도처리96 h적조망솔、천막세포수급조망상관기인mRNA균우우48 h（ P＜0．05）。결론 BRD4억제제GSK525762A대비인암CNE?2세포증식유독성작용，가유도CNE?2세포조망，회복조망상관기인적표체，병강저세포적침습능력。
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.
