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1,王亚南2   

  1. 1. 中国医学科学院基础医学研究所
    2. 中国医学科学院基础医学研究所&北京协和医学院基础医学院
  • 收稿日期:2020-12-07 修回日期:2021-04-25 出版日期:2021-07-05 发布日期:2021-06-17
  • 通讯作者: 王亚南 E-mail:shlwangyanan@gmail.com
  • 基金资助:
    国家自然科学基金;中国医学科学院医学与健康科技创新工程经费资助

Sodium hydroxide enhances the efficacy of rapamycin to suppress proliferation of embryonic fibroblasts in an animal model of Tsc2 null mouse

  • Received:2020-12-07 Revised:2021-04-25 Online:2021-07-05 Published:2021-06-17

摘要: 目的:研究氢氧化钠与雷帕霉素(Rapa)单独或联合处理对Tsc2基因缺失小鼠胚胎成纤维细胞的影响。方法:将Tsc2-/- 细胞及其对照的野生型(WT)细胞按照处理方式分为:对照组,氢氧化钠处理组(NaOH),雷帕霉素处理组(Rapa)和氢氧化钠、雷帕霉素联合处理组(Combo)。使用MTT法测定细胞增殖速率;克隆形成测定细胞集落形成能力;PI染色流式细胞术检测细胞周期;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组的两组Tsc2-/-细胞均表现为增殖速率降低、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NaOH组和Rapa组相比,Combo组的Tsc2-/-细胞在增殖速率和克隆形成数上进一步降低,G1期细胞的比例进一步升高(P <0.05),但p-S6和p-4EBP1的蛋白表达水平只显著低于NaOH组(P <0.05)。结论:氢氧化钠增强了Rapa抑制 Tsc2-/- 细胞增殖的效果。

关键词: 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 and rapamycin alone or combination in treating Tsc2-/- cells. Methods The wild type and Tsc2-/- cells were treated with sodium hydroxide and/or rapamycin and divided into 4 groups: control, NaOH, rapamycin (Rapa), and combination (Combo) groups. Cell proliferation was measured by MTT assay and colony formation. Flow cytometry was used to detect the influence of cell cycle. The protein expression levels of S6, p-S6, p-4EBP1 were detected by Western Blot. Results Compared with control group, both Tsc2-/- cells in NaOH and Rapa groups demonstrated the reduced proliferation rate and colonies number (P <0.05); additionally, only in Rapa group, the percentage of Tsc2-/- cells in G1 phase increased and the protein levels of p-S6 and p-4EBP1 decreased (P <0.05). Compared with NaOH and Rapa groups, Tsc2-/- cells in Combo group had a further decreased in proliferation rate and colonies number, and the increased G1 phase percentage (P <0.05). But the p-S6 and p-4EBP1 levels of Tsc2-/- cells in Combo group were only significantly lower than those in NaOH group (P <0.05). Conclusion Sodium hydroxide could enhance the efficacy of rapamycin in suppressing Tsc2-/- cells proliferation.

Key words: Keywords: Tuberous sclerosis complex, mTOR, tumor microenvironment, Rapamycin

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