摘要
目的研究二烯丙基二硫(diallyl disulfide,DADS)对人恶性胶质瘤U251细胞增殖及细胞周期的影响。方法采用MTT法、流式细胞术等方法检测DADS对U251细胞的增殖抑制及细胞周期的影响。免疫细胞化学、qRT-PCR与Western blot检测cyclin B1与c-Myc表达。结果 MTT法显示,15、30、45、60 mg/L DADS处理U251细胞48小时后,生长抑制率分别为22.0%、38.8%、49.2%、55.3%,与对照组比较差异有统计学意义(P<0.05)。流式细胞术结果显示,15、30、45、60 mg/L DADS作用U251细胞48小时后,G2/M期细胞分别为(14.1±1.2)%、(25.0±0.6)%、(41.2±1.4)%、(53.5±3.3)%,较对照组(9.6±0.9)%差异具有统计学意义(P<0.05)。免疫细胞化学显示,45 mg/L DADS处理U251细胞48小时后,cyclin B1与c-Myc蛋白平均光密度值分别从0.52±0.09、0.56±0.08显著下降至0.23±0.02、0.39±0.04(P<0.05)。qRT-PCR结果显示,cyclin B1与c-Myc mRNA表达分别下调42.0%与63.0%(P<0.05)。Western blot结果显示,cyclin B1、c-Myc蛋白与β-actin的平均灰度值比分别为1.12±0.14与1.20±0.15,明显低于对照组1.74±0.15与2.23±0.19(P<0.05)。结论 DADS可明显抑制恶性胶质瘤U251细胞增殖与阻滞G2/M期,机制与下调cyclin B1、c-Myc有关。
Objective To investigate the effect of diallyl disulfide （DADS） on the proliferation and cell cycle of human glioblastoma U251 cells. Methods Human glioblastoma U251 cells were treated with DADS in vitro. The inhibition of U251 cell growth was measured by MTT. The effect of DADS on the cell cycle of U251 cells was analyzed by flow cytometry. Expression of cyclin B1 and c-Myc was determined by immunocytochemistry, qRT-PCR and Western blot. Results MTF assay showed that the treatment of 15,30,45,60 mg/L DADS for 48 h significantly inhibited the proliferation of U251 cells with inhibition rates of 22. 0% ,38. 8% ,49. 2% and 55.3%, respectively （P 〈 0. 05 ）. Flow cytometry analysis revealed that U251 cells treated with 15,30,45,60 mg/L DADS for 48 h were significantly arrested in GJM phase [ （14. 1 ± 1.2） %, （ 25.0 ± 0. 6） %, （41.2 ± 1.4） % and （53.5 ± 3.3 ） %, respectively ], compared to untreated ceils [ （9.6 ± 0. 9 ） % ,P 〈 0.051. Immuocytochemistry revealed that the expression of cyclin B1 and c-Myc proteins in U251 cells treated with 45 mg/L DADS for 48 h decreased from 0. 52 ± 0. 09 and 0. 56 ± 0.08 to 0. 23 ± 0. 02 and 0. 39 ± 0. 04 （ P 〈 0. 05 ）. qRT-PCR showed that the expression of cyclin B1 and c-Myc mRNA decreased by 42.0% and 63.0% ,respectively （P 〈 0. 05 ）. Western blot exhibited that the expression of cyclin B1 and c-Myc proteins in DADS-treated cells was 1.12 ± 0. 14 and 1.20 ± 0. 15, which was lower than that of untreated cells （ 1.74 ± 0. 15 and 2. 23 ± 0. 19 ; P 〈 0. 05 ）. Conclusion DADS can inhibit the proliferation of human glioblastoma U251 cells and induce cycle arrest in GJM phase in a dose- dependent manner,which is associated with the down-regulation of cyclin B1 and c-Myc expression.
出处
《实用肿瘤杂志》
CAS
2014年第2期126-132,共7页
Journal of Practical Oncology
基金
国家自然科学基金(81102854
31100935)
湖南省卫生厅科研课题计划项目(B2009057)
关键词
神经胶质瘤
病理学
二硫化物类
药理学
烯丙化合物
药理学
细胞周期
药物作用
G2期
流式细胞术
glioma/pathology
disulfides/pharmacology
allyl compounds/pharmacology
cell cycle/drug effects
G2 phase
cell proliferation/drug effects
flow cytometry
作者简介
汪雪菁（1971-），女，湖南衡阳人，副主任医师，硕士，从事神经内科学研究。
通信作者E-mail：suqil945@163．com
