解剖学报

解剖学报 ›› 2022, Vol. 53 ›› Issue (3): 323-329.doi: 10.16098/j.issn.0529-1356.2022.03.008

• 肿瘤生物学 • 上一篇    下一篇

咪喹莫特下调STAT3/核因子κB信号通路抑制脑胶质细胞瘤U87细胞的增殖

王澍1 尹璐1 刘宏斌1 徐加志1 赵吉波1 潘云志1 孙玉荣2*   

  1. 1.齐齐哈尔医学院附属第三医院神经内科; 2.齐齐哈尔医学院病理学院，黑龙江 齐齐哈尔 161006
  • 收稿日期:2020-08-16 修回日期:2020-12-25 出版日期:2022-06-06 发布日期:2019-06-06
  • 通讯作者: 孙玉荣 E-mail:343190863@qq.com
  • 基金资助:
    齐齐哈尔医学科学院临床科研基金项目

Imiquimod inhibiting the proliferation of gliocytoma U87 cells by down-regulating STAT3/nuclear factor-κB signal pathway

WANG  Shu1  YIN  Lu1  LIU  Hong-bin1  XU  Jia-zhi1  ZHAO  Ji-bo1  PAN  Yun-zhi1  SUN  Yu-rong2*   

  1. 1.Department of Neurology, the Third Affiliated Hospital of Qiqihar Medical College, Heilongjiang Qiqihar 161006, China; 2.School of Pathology, Qiqihar Medical College, Heilongjiang Qiqihar 161006, China
  • Received:2020-08-16 Revised:2020-12-25 Online:2022-06-06 Published:2019-06-06
  • Contact: SUN Yu-rong E-mail:343190863@qq.com

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摘要：

目的 探讨咪喹莫特（IMQ）对脑胶质细胞瘤U87细胞增殖的影响。  方法 U87细胞分为对照组、1mmol/L IMQ组、5mmol/L IMQ组、1 mmol/L IMQ+STAT3抑制剂（inhibitor)（STAT3-IN）组和5 mmol/L IMQ+STAT3-IN组。5-乙炔基-2’-脱氧尿苷（EdU）和MTT实验检测各组U87细胞EdU标记的细胞数目或增殖吸光度值；Real-time PCR、ELISA检测各组U87细胞白细胞介素6（IL-6）mRNA、肿瘤坏死因子-α（TNF-α）mRNA及其蛋白含量；Western blotting检测各组U87细胞STAT3、磷酸化STAT3（p-STAT3）、核因子（NF）-κB、磷酸化NF-κB（p-NF-κB）蛋白表达。  结果 相对于对照组，1mmol/L IMQ组和5mmol/L IMQ组U87细胞的EdU标记细胞数目和吸光度值依次降低，并呈现剂量依赖性（P＜0.01, n=10），IMQ+STAT3-IN组和5 mmol/L IMQ+STAT3-IN组EdU标记的U87细胞数目、吸光度值均降低。1mmol/L IMQ组和5mmol/L IMQ组U87细胞STAT3、p-STAT3、NF-κB、p-NF-κB或IL-6、TNF-α蛋白均比对照组低表达（P＜0.01, n=10），1  mmol/L IMQ+STAT3-IN组和5 mmol/L IMQ+STAT3-IN组上述蛋白持续低表达（P＜0.01, n=10）。  结论 咪喹莫特通过下调STAT3/NF-κB通路降低IL-6、TNF-α含量，进而抑制U87细胞增殖。

关键词: 咪喹莫特, 脑胶质细胞瘤, 细胞增殖, 实时定量聚合酶链反应, 人

Abstract:

Objective  To explore the effect of imiquimod (IMQ) on the proliferation of  glioma U87 cell line U87.   Methods  U87 cells were divided into control group, 1 mmol/L I mmol/L MQ group, 5mmol/L IMQ group, 1 mmol/L IMQ+STAT3 inhibitor（STAT3-IN） group and 5 mmol/L IMQ+STAT3-IN group. To detect the number of 5-ethynyl-2’-deoxyuridine(EdU)-labeled cells or proliferation absorbance(A) values in each group by EdU and MTT assays. Interleukin(IL)-6 mRNA, tumor necrosis factor(TNF)-α mRNA and protein content in U87 cells of each group were detected by Real-time PCR or ELISA. Western blotting was used to detect the protein expression of STAT3, phosphorylated STAT3 (p-STAT3), nuclear factor(NF)-κB and phosphorylated NF-κB (p-NF-κB) in U87 cells of each group.  Results  Compared with the control group, the number of EdU-labeled cells and absorbance values of U87 cells were successively decreased in 1mmol/L IMQ group and 5mmol/L IMQ group, showing a dose-dependent manner (P＜0.01, n=10). However, the number of EdU-labeled cells and the A values in IMQ+STAT3-IN group and 5 mmol/L IMQ+STAT3-IN group were significantly reduced. Compared with the control group, the protein expression of STAT3, p-STAT3, NF-κB, p-NF-κB, IL-6 and TNF-α were continuing low level in U87 cells of 1mmol/L IMQ group and 5mmol/L IMQ group (P＜0.01, n=10). As well as in 1 mmol/L IMQ+STAT3-INgroup and 5 mmol/L IMQ+STAT3-IN group, the proteins of above were low expressed (P＜0.01, n=10).   Conclusion Imiquimod decreased the contents of IL-6 and TNF-α by down-regulating STAT3/NF-κB pathway, and thus inhibited the proliferation of U87 cells.

Key words: Imiquimod, Gliocytoma, Cell proliferation, Real-time PCR, Human

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