天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (11): 1126-1132.doi: 10.11958/20210976

• 细胞与分子生物学 • 上一篇    下一篇

尿石素C对胶质母细胞瘤生物学行为的调控作用及机制

刘翠兰，李建军，赵娣，李承龙，邱长云，刘松   

  1. 1滨州医学院附属医院代谢与神经精神疾病研究所（邮编256603）；2济南市第一人民医院急诊科；3滨州医学院附属医院神经外科，4甲状腺乳腺外科
  • 收稿日期:2021-04-25 修回日期:2021-06-16 出版日期:2021-11-15 发布日期:2021-11-19
  • 基金资助:
    山东省自然科学基金（ZR2014HQ020）；山东省医药卫生科技发展计划项目（2019WS323、202003090720）

The regulatory effects and mechanism of urolithin C on biological behaviors of glioblastoma

LIU Cui-lan, LI Jian-jun, ZHAO Di, LI Cheng-long, QIU Chang-yun, LIU Song   

  1. 1 Institute of Metabolism and Neuropsychiatry, Binzhou Medical University Hospital, Binzhou 256603， China; 2 Department of Emergency, Jinan First People's Hospital; 3 Department of Neurosurgery, 4 Thyroid and Breast Surgery, Binzhou Medical University Hospital
  • Received:2021-04-25 Revised:2021-06-16 Published:2021-11-15 Online:2021-11-19

PDF (PC)

2931

摘要： 目的　探讨尿石素C（UC）对胶质母细胞瘤（GBM）U251和U87 MG细胞生物学行为的影响及调控机制。方法　将U251和U87 MG细胞分为0、20、40、80、120和160 μmol/L UC处理组，通过CCK-8法分别检测各组细胞24、48和72 h时增殖率。将细胞分为0、40、80和120 μmol/L UC处理组，通过细胞克隆形成实验检测细胞克隆形成能力，划痕愈合实验检测24 h和48 h时细胞迁移能力，Transwell小室侵袭实验检测24 h时细胞侵袭能力，流式细胞术检测24 h时细胞凋亡率及细胞周期，Western blot检测AMP依赖的蛋白激酶（AMPK）、细胞外调节蛋白激酶（ERK）和丝裂原活化蛋白激酶p38（p38 MAPK）蛋白表达水平和磷酸化水平。结果　与0 μmol/L组比较，U251细胞中不同浓度UC组于48 h开始增殖率均降低，而U87 MG细胞中40 μmol/L及以上浓度组于48 h时增殖率开始降低（P＜0.05）。与0 μmol/L组比较，40、80和120 μmol/L组U251和U87 MG细胞的克隆形成率、24 h和48 h时的细胞迁移能力和24 h时的细胞侵袭能力降低，且均呈浓度依赖性（P＜0.05）。与0 μmol/L组比较，120 μmol/L组U251和U87 MG细胞凋亡率均增加，40、80和120 μmol/L组U251和U87 MG细胞周期均阻滞在S期（P＜0.05）。与0 μmol/L组比较，40、80和120 μmol/L组U251和U87 MG细胞中p-AMPK和p-p38 MAPK水平均升高，80和120 μmol/L组U251细胞中p-ERK水平升高，但仅120 μmol/L组U87 MG细胞p-ERK水平升高（P＜0.05）。结论　一定浓度的UC可抑制GBM细胞增殖、迁移和侵袭，诱导肿瘤细胞凋亡和细胞周期阻滞，其机制可能与调控AMPK/ERK/p38 MAPK信号通路有关。

关键词: 胶质母细胞瘤, 细胞系, 肿瘤, 细胞增殖, 细胞运动, 细胞周期, 细胞凋亡, 尿石素C, 细胞侵袭

Abstract: Objective　To investigate the effects of urolithin C (UC) on biological behaviours of glioblastoma (GBM) U251 and U87 MG cells and its regulatory mechanism. Methods　U251 and U87 MG cells were divided into 0, 20, 40, 80, 120 and 160 μmol/L UC treatment groups. The cell proliferation rates in each group at 24, 48 and 72 h were detected by CCK-8 method, respectively. The cells were divided into 0, 40, 80 and 120 μmol/L UC treatment groups. The cell colony formation ability was detected by colony formation assay. The cell migration ability at 24 and 48 h was detected by wound healing assay. The cell invasion ability was detected by Transwell invasion assay at 24 h, and the cell apoptosis and cell cycle were detected by flow cytometry at 24 h. Western blot assay was used to detect the expression levels and phosphorylation levels of AMPK, ERK and p38 MAPK. Results Compared with the 0 μmol/L group, the proliferation rates of U251 cells decreased at 48 h at different concentration UC groups, while the proliferation rate of U87 MG cells at 40 µmol/L and above decreased at 48 h (P＜0.05). Compared with the 0 µmol/L group, the clone formation rate, cell migration and invasion ability decreased in a concentration-dependent manner in the 40, 80 and 120 μmol/L U251 and the U87 MG cell groups (P＜0.05). Compared with the 0 μmol/L group, the apoptosis rates increased in the 120 µmol/L U251 and the U87 MG cell groups, and the cell cycle of U251 and U87 MG cells in the 40, 80 and 120 μmol/L groups were all blocked in the S phase (P＜0.05). Compared with the 0 μmol/L group, the levels of p-AMPK and p-p38 MAPK in U251 and U87 MG cells increased in the 40, 80, and 120 μmol/L groups, and the p-ERK levels in U251 cells increased in the 80 and 120 μmol/L groups, but the p-ERK level of U87 MG cells only increased in the 120 μmol/L group (P＜0.05). Conclusion　A certain concentration of UC can inhibit the proliferation, migration and invasion of GBM cells, induce cell apoptosis and cell cycle arrest, and the mechanism may be related to the regulation of AMPK/ERK/p38 MAPK signaling pathway.

Key words: glioblastoma, cell line, tumor, cell proliferation, cell movement, cell cycle, apoptosis, urolithin C, cell invasion