中国癌症杂志 ›› 2015, Vol. 25 ›› Issue (11): 900-904.doi: 10.3969/j.issn.1007-3969.2015.11.010

• 论著 • 上一篇    下一篇

紫檀芪诱导人视网膜母细胞瘤WERI-Rb-1细胞的细胞凋亡及其机制研究

张 静1,沈永青2,智利彩1,常 亮3,仇 炜4,5   

  1. 1. 石家庄市第四医院产前诊断中心,河北 石家庄 050011 ;
    2. 河北中医学院护理学院,河北 石家庄 050020 ;
    3. 北京大学第三医院妇产科,北京 100191 ;
    4. 石家庄市鹿泉区人民医院外科,河北 石家庄 050020 ;
    5. 首都医科大学附属北京友谊医院泌尿外科,北京 100050
  • 出版日期:2015-11-30 发布日期:2016-02-01
  • 通信作者: 仇炜 E-mail:qiuwei618@163.com
  • 基金资助:
    河北省医学科学研究重点课题(ZD20140032)。

Pterostilbene induces retinoblastoma WERI-Rb-1 cell apoptosis via autophagy induction

ZHANG Jing1, SHEN Yongqing2, ZHI Licai1, CHANG Liang3, QIU Wei4,5   

  1. 1.Prenatal Diagnosis Center, Fourth Hospital of Shijiazhuang, Shijiazhuang 050011, Hebei, China; 2.College of Nursing, Hebei University of Chinese Medicine, Shijiazhuang 050200, Hebei, China; 3.Department of Gynaecology and Obstetrics, Peking University Third Hospital, Beijing 100191, China; 4.Department of Surgery, the People’s Hospital of Luquan District, Shijiazhuang 050200, Hebei, China; 5.Department of Urology, Beijing Friendship Hospital, Beijing 100050, China
  • Published:2015-11-30 Online:2016-02-01
  • Contact: QIU Wei E-mail: qiuwei618@163.com

摘要:

背景与目的:紫檀芪是一种天然抗氧化剂,其抗视网膜母细胞瘤的效果仍不明确。拟探讨紫檀芪对人视网膜母细胞瘤WERI-Rb-1细胞的细胞增殖、细胞凋亡及细胞自噬的作用,并初步阐明其作用机制。方法:采用细胞计数试剂盒(cell counting kit-8,CCK-8)法测定细胞的增殖活力,Annexin V/PI流式细胞术检测细胞凋亡,吖啶橙染色观察细胞内自噬囊泡的变化,蛋白[质]印迹法(Western blot)检测LC3B及P62蛋白的表达。结果:紫檀芪显著抑制WERI-Rb-1细胞的增殖活力(P<0.01),以25、50和100 μmol/L的药物浓度作用细胞24 h时,细胞增殖活力分别为(93.02±0.47)%、(55.10±2.04)%和(30.33±1.45)%;50 μmol/L紫檀芪处理细胞12、24和48 h时,细胞增殖活力分别为(88.38±3.70)%、(53.37±1.17)%和(29.60±1.05)%。紫檀芪显著诱导细胞凋亡(P<0.01),对照组、24和48 h细胞的凋亡率分别为(4.08±0.79)%、(13.44±2.12)%和(23.49±2.01)%。紫檀芪能够激活WERI-Rb-1细胞发生细胞自噬,上调LC3蛋白的表达,下调P62蛋白的表达,增加细胞内自噬囊泡的数量(P<0.01)。3-MA及Beclin1 siRNA抑制细胞自噬后能够部分逆转紫檀芪的抗肿瘤作用(P<0.01)。3-MA抑制细胞自噬后,紫檀芪处理组的细胞凋亡率为(12.97±2.09)%,3-MA+紫檀芪组为(8.35±1.11)%。siRNA敲低Beclin1后,紫檀芪处理组和siRNA+紫檀芪组的细胞凋亡率分别为(13.80±2.19)%和(9.62±0.52)%。结论:紫檀芪可以显著抑制WERI-Rb-1细胞的细胞增殖并激活细胞自噬促进细胞凋亡。

关键词: 紫檀芪, 视网膜母细胞瘤, 细胞凋亡, 细胞增殖, 自噬

Abstract: Background and purpose: Pterostilbene is a natural antioxidant, whose role in retinoblastoma remains unclear. The aim of this study is to probe the effects of pterostilbene on the proliferation, apoptosis and autophagy in retinoblastoma WERI-Rb-1 cell lines. Methods: Cell counting kit-8 (CCK-8) assays were used to analyze the effects of pterostilbene on the proliferation of WERI-Rb-1 cells. Apoptosis rate was determined by Annexin V/PI. Autophagic vacuoles were observed by acridine orange staining. LC3 and P62 protein expressions were determined using Western blot. Results: Pterostilbene significantly inhibited the proliferation of WERI-Rb-1 cells (P<0.01). The cell viability were (93.02±0.47)%, (55.10±2.04)% and (30.33±1.45)% after WERI-Rb-1 cells were treated with 25, 50 and 100 μmol/L pterostilbene for 24 h, and the cell viability were (88.38±3.70)%, (53.37±1.17)%, (29.60±1.05)% after WERI-Rb-1 cells were treated with 50 μmol/L pterostilbene for 12, 24 and 48 h. Pterostilbene induced cell apoptosis (P<0.01), the apoptosis rates of control group, 24 h treated group and 48 h treated group were (4.08±0.79)%, (13.44±2.12)% and (23.49±2.01)%. Pterostilbene induced autophagy of WERI-Rb-1 cells, increased LC3 expression, downregulated P62 expression and increased the number of autophagic vacuoles in WERI-Rb-1 cells (P<0.01). 3-MA and Beclin1 were able to rescue pterostilbene-induced cell death (P<0.01). After 3-MA was used to blunt autophagosome formation, the apoptosis rate markedly decreased in 3-MA+pterostilbene-treated cells compared with cells treated with pterostilbene alone [(12.97±2.09)% vs (8.35±1.11)%], and after siRNA was used to knockdown Beclin1, the apoptosis rate had the same change [(13.80±2.19)% vs (9.62±0.52)%]. Conclusion: Pterostilbene can inhibit the proliferation of WERI-Rb-1 cells and induce cell apoptosis via autophagy activation.

Key words: Pterostilbene, Retinoblastoma, Cell apoptosis, Cell proliferation, Cell autophagy