吉林大学学报(医学版) ›› 2021, Vol. 47 ›› Issue (3): 566-574.doi: 10.13481/j.1671-587X.20210305

• 基础研究 • 上一篇    下一篇

尿石素B对人神经胶质瘤U118 MG细胞生物学行为的影响及其机制

刘翠兰1,李建军2,姜贺1,刘晶1,王丹1,李晨1(),赵娣1()   

  1. 1.滨州医学院附属医院代谢与神经精神疾病研究所,山东 滨州 256603
    2.山东省 济南市 第一人民医院急诊科,山东 济南 250000
  • 收稿日期:2020-11-25 出版日期:2021-05-28 发布日期:2021-05-28
  • 通讯作者: 李晨,赵娣 E-mail:lc_0625@163.com;zhaodi914@126.com
  • 作者简介:刘翠兰(1987-),女,山东省滨州市人,主管技师,理学硕士,主要从事胶质细胞瘤发病机制方面的研究。
  • 基金资助:
    国家自然科学基金项目(81601189);滨州医学院科研计划与科研启动基金项目(BY2019KJ02)

Effect of urolithin B on biological behaviors of human glioblastome U118 MG cells and its mechanism

Cuilan LIU1,Jianjun LI2,He JIANG1,Jing LIU1,Dan WANG1,Chen LI1(),Di ZHAO1()   

  1. 1.Institute for Metabolic and Neuropsychiatric Disorders,Binzhou Medical University Hospital,Binzhou 256603,China
    2.Department of Emergency,First People’s Hospital of Jinan,Shandong Province,Jinan 250000,China
  • Received:2020-11-25 Online:2021-05-28 Published:2021-05-28
  • Contact: Chen LI,Di ZHAO E-mail:lc_0625@163.com;zhaodi914@126.com

摘要: 目的

观察尿石素B(UB)对人神经胶质瘤(GBM)的U118 MG细胞增殖、迁移、侵袭和凋亡的影响,并探讨其作用机制。

方法

体外培养U118 MG细胞,将细胞分为对照组(0 μmol·L-1 UB)和不同浓度(20、40、80、120、160和200 μmol·L-1)UB组,培养24、48和72 h后,采用CCK-8法检测各组细胞增殖率。将U118 MG细胞分为对照组(0 μmol·L-1 UB)和不同浓度(40、80和120 μmol·L-1)UB组,采用细胞克隆形成实验检测各组U118 MG细胞克隆形成率,划痕愈合实验检测各组U118 MG细胞划痕愈合率,Transwell小室实验检测各组U118 MG细胞侵袭百分率,流式细胞术检测各组U118 MG细胞凋亡率和不同细胞周期U228 MG细胞百分率,Western blotting法检测各组U118 MG细胞中波形蛋白(Vimentin)、上皮型钙黏附蛋白(E-cadherin)、B细胞淋巴瘤2(Bcl-2)和Bcl-2相关X蛋白(Bax)表达水平。

结果

CCK-8法检测,作用24、48和72 h后,与对照组比较,不同浓度UB组U118 MG细胞增殖率明显降低(P<0.01),且呈浓度和时间依赖性。细胞克隆形成实验,与对照组比较,40、80和120 μmol·L-1 UB组U118 MG细胞克隆形成率降低(P<0.01),且呈浓度依赖性;划痕愈合实验,作用24和48 h后,与对照组比较,40、80和120 μmol·L-1 UB组U118 MG细胞划痕愈合率明显降低(P<0.05或P<0.01),且呈浓度依赖性;Transwell小室实验,与对照组比较,40、80和120 μmol·L-1 UB组侵袭U118 MG细胞百分率明显降低(P<0.05或P<0.01),且呈浓度依赖性;流式细胞术,与对照组比较,80和120 μmol·L-1 UB组U118 MG细胞凋亡率升高(P<0.05或P<0.01),且呈浓度依赖性,G2/M期细胞百分率升高(P<0.05或P<0.01);Western blotting法检测,与对照组比较,不同浓度UB组U118 MG细胞中Vimentin和Bcl-2蛋白表达水平降低(P<0.05或P<0.01),E-cadherin和Bax蛋白表达水平升高(P<0.01)。

结论

UB能够抑制人GBM的U118 MG细胞增殖、迁移和侵袭,抑制U118 MG细胞中Bcl-2蛋白表达并上调Bax蛋白表达,诱导细胞凋亡,并引起细胞周期G2/M期阻滞。

关键词: 尿石素B, 神经胶质瘤, 细胞增殖, 细胞迁移, 细胞侵袭, 细胞凋亡

Abstract: Objective

To observe the effect of urolithin B (UB) on the proliferation, migration, invasion and apoptosis of human glioblastoma(GBM)U118 MG cells,and to discuss its mechanism.

Methods

The U118 MG cells were cultured in vitro and divided into control group(0 μmol·L-1 UB) and different concentrations (20, 40, 80, 120, 160 and 200 μmol·L-1) of UB groups. After cultured for 24, 48 and 72 h, the proliferation rates of U118 MG cells in various groups were detected by CCK-8 method.The U118 MG cells were divided into control group(0 μmol·L-1 UB) and different concentrations (40, 80 and 120 μmol·L-1) of UB groups,and clone formation experiment was used to detect the rates of clone formation of U118 MG cells in various groups; cell scratch healing test was used to detect the scratch healing rates of U118 MG cells in various groups; Transwell chamber assay was used to detect the percentages of invasion U118 MG cells in various groups; flow cytometry was used to detect the apoptotic rates of U118 MG cells and the percentages of U118 MG cells at different cell cycles in various groups; Western blotting method was used to detect the expression levels of Vimentin,epithelial adhesive protein(E-cadherin),B cell lymphoma-2(Bcl-2) ,and Bcl-2 related X protein(Bax) in the U118 MG cells in various groups.

Results

The CCK-8 results showed that compared with control group, the proliferation rates of U118 MG cells in different concentrations of UB groups at 24, 48 and 72 h after treatment were decreased(P<0.01) in a dose and time-dependent manner. The clone formation experiment results showed that compared with control group, the rates of clone formation of U118 MG cells in 40, 80 and 120 μmol·L-1 UB groups were decreased(P<0.01) in a dose-dependent manner.The cell scratch test results showed that compared with control group, the scratch healing rates of U118 MG cells in 40, 80 and 120 μmol·L-1 UB groups at 24 and 48 h after treatment were significantly decreased(P<0.05 or P<0.01) in a dose-dependent manner.The Transwell chamber assay results showed that compared with control group, the percentages of invasion U118 MG cells in 40, 80 and 120 μmol·L-1 UB groups were decreased significantly(P<0.05 or P<0.01) in a dose- dependent manner.The flow cytometry results showed that compared with control group, the apoptotic rates of U118 MG cells in 80 and 120 μmol·L-1 UB groups were increased(P<0.05 or P<0.01) in a dose-dependent manner, and the percentages of U118 MG cells at G2/M phase were increased (P<0.01). The Western blotting results showed that compared with control group, the expression levels of Vimentin and Bcl-2 proteins in U118 MG cells in different concentrations of UB groups were decreased (P<0.05 or P<0.01),and the expression levels of E-cadherin and Bax proteins were increased (P<0.01).

Conclusion

UB can inhibit the proliferation, migration and invasion of human GBM U118 MG cells, inhibit the expression of Bcl-2 protein, increase the expression of Bax protein, induce the apoptosis, and result in G2/M phase arrest.

Key words: urolithin B, glioblastoma, cell proliferation, cell migration, cell invasion, cell apoptosis

中图分类号: 

  • R739.41