Toosendanin inhibits proliferation and promotes apoptosis of gastrointestinal stromal tumor-T1 cells by down-regulating c-kit expression

Xiaoyu HU; Wensheng YANG; Zhengyu KANG; Zhengwei PENG; Ling LI; Xi LIU; Yijia CHEN; Lin ZHANG

doi:10.16016/j.1000-5404.202012232

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Toosendanin inhibits proliferation and promotes apoptosis of gastrointestinal stromal tumor-T1 cells by down-regulating c-kit expression

Basic Medicine | 更新时间：2026-01-09

- Toosendanin inhibits proliferation and promotes apoptosis of gastrointestinal stromal tumor-T1 cells by down-regulating c-kit expression
- Journal of Third Military Medical University Vol. 43, Issue 13, Pages: 1235-1242(2021)
- 作者机构：
  
  1.610031 成都，西南交通大学医学院
  2.610083 成都，中国人民解放军西部战区总医院全军普外中心胃肠病区
  3.646000 四川泸州，西南医科大学临床医学院
- 作者简介：
- 基金信息：
  
  the Project of Innovative Personnel Training Foundation of General Hospital of Western Theater Command(417325E)
- DOI：10.16016/j.1000-5404.202012232
  CLC： R285.5;R73-361;R730.262
- Received：29 December 2020，
  
  Revised：2021-03-02，
  
  Published：15 July 2021
- 稿件说明：
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Xiaoyu HU, Wensheng YANG, Zhengyu KANG, et al. Toosendanin inhibits proliferation and promotes apoptosis of gastrointestinal stromal tumor-T1 cells by down-regulating c-kit expression[J]. Journal of Third Military Medical University, 2021, 43(13): 1235-1242.
DOI：

Xiaoyu HU, Wensheng YANG, Zhengyu KANG, et al. Toosendanin inhibits proliferation and promotes apoptosis of gastrointestinal stromal tumor-T1 cells by down-regulating c-kit expression[J]. Journal of Third Military Medical University, 2021, 43(13): 1235-1242. DOI： 10.16016/j.1000-5404.202012232.

摘要

目的

研究川楝素(toosendanin，TSN)对胃肠间质瘤细胞增殖及凋亡的影响，并对其相关机制进行初步探索。

方法

不同浓度TSN体外干预GIST-T1细胞，采用CCK-8检测TSN对GIST-T1细胞增殖的影响；根据结果进一步选择实验浓度，分为0、15、30 nmol/L TSN组，50 nmol/L伊马替尼(imatinib，IM)作为阳性对照；流式细胞仪及Hoechst 33342/PI荧光双染检测细胞凋亡及细胞周期；qRT-PCR及Western blot检测c-KIT/PI3K/AKT/mTOR mRNA及相关蛋白表达。

结果

TSN可抑制GIST-T1细胞增殖(

0.01)，其48 h IC

为18.75 nmol/L，72 h IC

为17.93 nmol/L。与对照组相比，15、30 nmol/L TSN组，50 nmol/L IM均可引起不同程度细胞凋亡(

0.01)，但各药物组间凋亡率差异无统计学意义。30 nmol/L TSN可使细胞周期发生S期阻滞(

0.01)。qRT-PCR检测结果显示: 15 nmol/L TSN即可下调GIST-T1细胞c-KIT、PI3K、AKT、mTOR mRNA表达(

0.05，

0.01)。Western blot结果也证实: 15 nmol/L TSN同样抑制了c-KIT、PI3K、AKT、mTOR及其磷酸化蛋白的表达(

0.05，

0.01)。

结论

TSN通过下调c-KIT表达，抑制GIST-T1细胞增殖并促进凋亡，可能是通过PI3K/AKT/mTOR通路发挥作用。

Abstract

Objective

To investigate the effects of toosendanin (TSN) on the proliferation and apoptosis of gastrointestinal stromal tumor (GIST) cells

and to explore the underlying mechanisms.

Methods

GIST-T1 cells were treated with TSN at different concentrations

and then CCK-8 assay was used to detect the effects of TSN on the proliferation of GIST-T1 cells. Subsequently

the experimental concentrations were further divided into 0

30 nmol/L TSN groups

with 50 nmol/L imatinib (IM) as the positive control. Flow cytometry and Hoechst 33342/PI fluorescence double staining were performed to detect cell apoptosis and cell cycle. Finally

the expression of c-KIT

PI3K

AKT and mTOR at mRNA and protein levels were determined by qRT-PCR and Western blotting.

Results

TSN inhibited the proliferation of GIST-T1 cells (

0.01)

and its IC

was 18.75 nmol/L at 48 h and 17.93 nmol/L at 72 h. As compared with the control group

the 15 and 30 nmol/L TSN groups as well as IM group induced cell apoptosis (

0.01)

though the apoptotic rate showed no significance among different drug groups; and 30 nmol/L TSN induced S-phase arrest (

0.01). In addition

qRT-PCR results suggested that 15 nmol/L TSN down-regulated the mRNA expression of c-KIT

PI3K

AKT and mTOR (

0.05

0.01)

and Western blotting confirmed the inhibitory effect of 15 nmol/L TSN on the expression of these proteins and their phosphorylated proteins (

0.05

0.01).

Conclusion

TSN inhibits the proliferation of GIST-T1 cells and promotes apoptosis by down-regulating c-KIT expression

which may function through PI3K/AKT/mTOR pathway.

关键词

Keywords

references

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