PUMA介导大蒜素诱导人卵巢癌耐顺铂SKOV-3/DDP细胞凋亡  

PUMA mediates human ovarian cancer SKOV-3/DDP cells apoptosis induced by diallyl trisulfide

作　　者：吴金兰[1,2] 万慧芳[1] 涂硕[1] 朱伟锋[1] 余乐涵[1] 万福生[1] 

机构地区：[1]南昌大学医学院生化与分子生物学教研室,江西南昌330006 [2]南昌大学第四附属医院检验科,江西南昌330006

出　　处：《中成药》2014年第8期1573-1579,共7页Chinese Traditional Patent Medicine

基　　金：江西省科技支撑计划重点资助项目(2009JX01081)

摘　　要：目的探讨p53上调凋亡调控因子(PUMA)在大蒜素诱导人上皮性卵巢癌耐顺铂细胞株(SKOV-3/DDP)细胞凋亡中的作用。方法将SKOV-3/DDP细胞随机分为4组:正常对照组、大蒜素组(于培养基中加入终质量浓度为20μg/mL大蒜素作用细胞48 h)、顺铂组(5μg/mL)、大蒜素+顺铂组。MTT法检测细胞存活率,荧光显微镜和流式细胞术检测细胞凋亡率,RT-PCR和Western-blot检测PUMA、bax及bcl-2的基因表达。结果与对照组比较,大蒜素组SKOV-3/DDP细胞凋亡率有明显的增加(P<0.05),且作用呈剂量依赖性,PUMA,Bax mRNA和蛋白表达上调,Bcl-2 mRNA和蛋白表达下调(P<0.05);与大蒜素组比较,顺铂+大蒜素组的细胞生长抑制率及凋亡率均有显著增加(P<0.05),Bax表达上调及Bcl-2表达下调的程度大于单用组(P<0.05);用特异性SiRNA沉默PUMA后,大蒜素组细胞凋亡率呈显著性下降(P<0.05)。结论 PUMA在介导大蒜素诱导SKOV-3/DDP细胞凋亡中发挥了关键性作用。AIM To investigate the effect of p53 up-regulated modulator of apoptosis protein （PUMA） on hu- man epithelial ovarian cancer SKOV-3/DDP cells apoptosis induced by diallyl trisulfide （DT）. METHODS Cul- tured SKOV-3/DDP cells were randomly divided into the control, DT-treated groups （20 μg/mL）, DDP-treated group （5 μg/mL）, DDP （5 μg/mL） ＋DT （20 μg/ml） group. After being incubated for 48 hours, the cell via- bility was detected by MTT. The apoptosis rate of cancer cells was detected by flow cytometric analysis. The mRNA and protein expressions of PUMA, bax and bcl-2 in the SKOV-3/DDP cells were detected by RT-PCR and Western blot. RESULTS Compared with the control, apoptosis rate of SKOV-3/DDP cells significantly increased in DT group in a dose-dependent manner （ P 〈 0. 05 ）, up-regulated the mRNA and protein expressions of PUMA and bax, and down-regulated the mRNA and protein expressions of bcl-2 （P 〈 0. 05）. Compared with the DT group, apoptosis rate of the cancer cells increased in DDP ＋ DT group （P 〈 0. 05 ）. After PUMA-specific interference of PUMA expression in the SKOV-3/DDP cells, the apoptosis rate of the cancer cells decreased obviously （P 〈 0.05）. CONCLUSION PUMA is critical for SKOV-3/DDP cell apoptosis induced by DT.

关 键 词：p53上调凋亡调控因子(PUMA) 大蒜素 SKOV-3/DDP细胞株 人卵巢癌 凋亡 

分 类 号：R966[医药卫生—药理学]