解毒三根汤对结肠癌SW480细胞侵袭转移及AKT/GSK-3β信号通路的影响

阮善明; 周萌萌; 袁莉; 沈敏鹤

doi:10.13288/j.11-2166/r.2019.16.012

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解毒三根汤对结肠癌SW480细胞侵袭转移及AKT/GSK-3β信号通路的影响

实验研究 | 更新时间：2021-11-10

- 解毒三根汤对结肠癌SW480细胞侵袭转移及AKT/GSK-3β信号通路的影响
- Effects of Jiedu Sangen Decoction（解毒三根汤） on Invasion and Metastasis of Colon Cancer SW480 Cells and AKT/GSK-3β Signaling Pathway
- 中医杂志 2019年60卷第16期页码：1401-1406
- 作者机构：
  
  1. 浙江中医药大学附属第一医院
  2. 浙江省衢州市人民医院
  3. 浙江中医药大学附属第一医院浙江省衢州市人民医院
- 作者简介：
- 基金信息：
  
  国家自然科学基金（81573902）;中国博士后科学基金（2017M612040,2018T110610）;浙江省自然科学基金（LY17H270007）;浙江省中医药科技计划项目（2015Z013）;浙江省重大科技专项计划（2014C03036）;中华中医药学会青年人才托举工程（QNRC2-C08）;浙江省高等学校中青年学科带头人培养对象项目（2017-248）;浙江省卫生高层次创新人才培养工程项目资助（2015-43）
- DOI：10.13288/j.11-2166/r.2019.16.012
  中图分类号： R285
- 纸质出版：2019
- 稿件说明：
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[1]阮善明,周萌萌,袁莉,沈敏鹤.解毒三根汤对结肠癌SW480细胞侵袭转移及AKT/GSK-3β信号通路的影响[J].中医杂志,2019,60(16):1401-1406.

RUAN Shanming, ZHOU Mengmeng, YUAN Li, et al. Effects of Jiedu Sangen Decoction（解毒三根汤） on Invasion and Metastasis of Colon Cancer SW480 Cells and AKT/GSK-3β Signaling Pathway[J]. Journal of Traditional Chinese Medicine2019, 60(16): 1401-1406.
[1]阮善明,周萌萌,袁莉,沈敏鹤.解毒三根汤对结肠癌SW480细胞侵袭转移及AKT/GSK-3β信号通路的影响[J].中医杂志,2019,60(16):1401-1406. DOI： 10.13288/j.11-2166/r.2019.16.012.

RUAN Shanming, ZHOU Mengmeng, YUAN Li, et al. Effects of Jiedu Sangen Decoction（解毒三根汤） on Invasion and Metastasis of Colon Cancer SW480 Cells and AKT/GSK-3β Signaling Pathway[J]. Journal of Traditional Chinese Medicine2019, 60(16): 1401-1406. DOI： 10.13288/j.11-2166/r.2019.16.012.

摘要

目的探讨解毒三根汤抗结肠癌侵袭转移的可能作用机制。方法在结肠癌SW480细胞中建立蛋白激酶B （AKT）敲减和相应阴性的细胞稳定株。实验分为阴性对照组、阴性三根汤组、AKT敲减对照组、AKT敲减三根汤组。首先用终浓度为50 ng/ml表皮细胞生长因子诱导各组细胞48 h建立上皮间质转化（EMT）模型。再用6 g/L解毒三根汤干预阴性三根汤组和AKT敲减三根汤组48 h。通过划痕实验、Transwell迁移和侵袭实验观察细胞侵袭转移能力

采用Western blot法检测EMT相关标记物（E-cadherin、N-cadherin、Vimentin、Occludin、Twist、Slug、Snail、ZEB1）及蛋白激酶B/糖原合成酶激酶3β（AKT/GSK-3β）信号通路相关蛋白表达。结果与阴性对照组比较

其余3组细胞12 h及24 h迁移距离均缩短

迁移及侵袭过膜细胞数均减少（P <0. 05）;与AKT敲减对照组比较

AKT敲减三根汤组细胞12 h及24 h迁移距离缩短

迁移及侵袭过膜细胞数减少（P <0. 05）。与阴性对照组比较

其余3组细胞E-cadherin、Occludin上调

N-cadherin、Vimentin、Slug、Twist、Snail、ZEB1、AKT、GSK-3β下调（P <0. 05）。与AKT敲减对照组比较

AKT敲减三根汤组E-cadherin、Occludin上调

N-cadherin、Vimentin、Snail、Slug、Twist、ZEB1、AKT、GSK-3β下调（P <0. 05）。结论解毒三根汤可能通过AKT/GSK-3β信号通路逆转结肠癌细胞的EMT

从而抗结肠癌侵袭转移。

Abstract

Objective To investigate the possible mechanism of Jiedu Sangen Decoction（解毒三根汤） against colon cancer invasion and metastasis. Methods Protein kinase B（ AKT） knockdown and corresponding negative cell stable strains were successfully established in colon cancer SW480 cells. The experiment was divided into a negative control group

a negative Sangen Decoction group

a AKT knockdown control group

and a AKT knockdown Sangen Decoction group. Epithelial-mesenchymal transition（ EMT） model was established by inducing epidermal growth factor at a final concentration of 50 ng/ml for 48 h. The 6 g/L Jiedu Sangen Decoction was used to intervene in the negative Sangen Decoction group and the AKT knockdown Jiedu Sangen Decoction group for 48 h. The invasion and metastasis ability of cells was observed by scratch test

Transwell migration and invasion assay. EMT-related markers（ E-cadherin

N-cadherin

Vimentin

Occludin

Twist

Slug

Snail

ZEB1） and protein kinase B/glycogen synthase kinase 3β（ AKT/GSK-3β） signaling pathway-associated protein expression were detected by Western blot. Results Compared with the negative control group

the migration distances of the other 3 groups were shortened at 12 h and 24 h

and the number of migration and invasion of membrane cells decreased（ P < 0. 05）. Compared with the AKT knockdown control group

the migration distance was shortened at 12 h and 24 h in AKT knockdown Sangen Decoction group

and the number of migration and invasion of membrane cells was decreased（ P < 0. 05）. Compared with the negative control group

the E-cadherin and Occludin was up-regulated

and N-cadherin

Vimentin

Slug

Twist

Snail

ZEB1

AKT and GSK-3β were down-regulated in the other 3 groups（ P < 0. 05）. Compared with the AKT knockdown control group

E-cadherin and Occludin were up-regulated

N-cadherin

Vimentin

Snail

Slug

Twist

ZEB1

AKT and GSK-3β were down-regulated in the AKT knockdown Sangen Decoction group（ P < 0. 05）.Conclusion Jiedu Sangen Decoction may reverse the EMT of colon cancer cells through AKT/GSK-3β signaling pathway

thus resisting colon cancer invasion and metastasis.

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