基础医学与临床 ›› 2021, Vol. 41 ›› Issue (7): 951-956.

• 研究论文 • 上一篇    下一篇

氢氧化钠增强雷帕霉素抑制Tsc2缺失的小鼠胚胎成纤维细胞的增殖

李凯, 王亚南*   

  1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 生理学系,北京 100005
  • 收稿日期:2020-12-07 修回日期:2021-04-27 出版日期:2021-07-05 发布日期:2021-06-17
  • 通讯作者: *shlwangyanan@gmail.com
  • 基金资助:
    国家自然科学基金(81602456)

Sodium hydroxide enhances the efficacy of rapamycin to suppress proliferation of Tsc2-null mouse embryonic fibroblasts

LI Kai, WANG Ya-nan*   

  1. Department of Physiology,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC,Beijing 100005,China
  • Received:2020-12-07 Revised:2021-04-27 Online:2021-07-05 Published:2021-06-17
  • Contact: *shlwangyanan@gmail.com

摘要: 目的 研究氢氧化钠(NaOH)与雷帕霉素(Rapa)单独或联合处理对Tsc2基因缺失(Tsc2-/-)小鼠胚胎成纤维细胞的影响。方法Tsc2-/- 细胞及其对照野生型(WT)细胞按照处理方式分为:对照组、NaOH处理组,Rapa处理组和NaOH+Rapa联合(Combo)处理组。采用MTT法检测细胞增殖能力;克隆形成实验检测细胞集落形成能力;流式细胞测量术检测细胞周期;免疫印迹法检测p-4EBP1、p-S6和S6蛋白表达水平。结果 与对照组相比,NaOH组和Rapa组的Tsc2-/-细胞均表现为增殖能力降低、克隆形成数减少(P<0.05);仅Rapa组的Tsc2-/-细胞G1期比例升高且p-S6和p-4EBP1的蛋白表达水平降低(P<0.05)。与单独的NaOH组和Rapa组相比,Combo组的Tsc2-/-细胞在增殖能力和克隆形成数量上呈进一步降低,G1期细胞的比例呈进一步升高(P<0.05),p-S6和p-4EBP1的蛋白表达水平低于NaOH组(P<0.05)。结论 氢氧化钠(15 μmol/L)能增强Rapa抑制 Tsc2-/- 细胞增殖的效果。

关键词: 结节性硬化症, mTOR, 肿瘤微环境, 雷帕霉素

Abstract: Objective To explore the effect of sodium hydroxide(NaOH) and rapamycin(Rapa) alone or their combination in treating Tsc2-null(Tsc2-/-) mouse embryonic fibroblasts. Methods The wild type and Tsc2-/- cells were treated with NaOH and/or Rapa and divided into 4 groups: control, NaOH group, Rapa group, and combination (Combo) group. Cell proliferation was measured by MTT assay and colony formation. Flow cytometry was used to detect the potential influence on cell cycle. The protein expression of S6, p-S6, p-4EBP1 was detected by Western blot. Results Compared to control group, Tsc2-/- cells in NaOH and Rapa groups both showed a reduced proliferation ability and colony counting (P<0.05); additionally, only in Rapa group, the percentage of Tsc2-/- cells in G1 phase increased and the protein level of p-S6 and p-4EBP1 all decreased (P<0.05). Compared with NaOH and Rapa groups, Tsc2-/- cells in Combo group had a further decreased in proliferation ability and colony number and an increased G1 phase percentage (P<0.05). But the level of p-S6 and p-4EBP1 of Tsc2-/- cells in Combo group was significantly lower than those in NaOH group (P<0.05). Conclusions NaoH(15 μmol/L) enhances the suppressing poliferation efficacy of Rapa in Tsc2-/- cells.

Key words: tuberous sclerosis complex, mTOR, tumor microenvironment, rapamycin

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