天然产物研究与开发 ›› 2023, Vol. 35 ›› Issue (7): 1135-1143.doi: 10.16333/j.1001-6880.2023.7.004

• 研究论文 • 上一篇    下一篇

基于PI3K/Akt/GSK-3β通路探讨蒲公英多糖对三阴性乳腺癌细胞迁移和侵袭的影响

刘晓燕1,龙   凤1,2,3*,赵玉1,侯   茜1,叶海琳1,周   旋1,李   雪1   

  1. 1甘肃中医药大学;2甘肃省中医药防治慢性疾病重点实验室;3甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室,兰州 730000
  • 出版日期:2023-07-31 发布日期:2023-07-31
  • 基金资助:
    甘肃省“双一流”科研重点项目(GSSYLXM-01);甘肃省高等学校创新基金项目(2021A-078)

Effect of dandelion polysaccharide on migration and invasion of triple-negative breast cancer cells based on PI3K/Akt/GSK-3β pathway

LIU Xiao-yan1,LONG Feng1,2,3*,ZHAO Yu1,HOU Qian1,YE Hai-lin1,ZHOU Xuan1,LI Xue1   

  1. 1Gansu University of Chinese Medicine;2Gansu Provincial Key Laboratory of Chinese Medicine for the Prevention and Treatment of Chronic Diseases;3Provincial-Level Key Laboratory for Molecular Medicine of Major Disease and Treatment with Traditional Chinese Medicine Research in Gansu Colleges and Universities,Lanzhou 730000,China
  • Online:2023-07-31 Published:2023-07-31

摘要:

本研究通过观察蒲公英多糖(dandelion polysaccharide,DP)对人三阴性乳腺癌MDA-MB-231细胞的增殖、迁移和侵袭的影响,探讨DP抑制乳腺癌细胞的分子机制。用DP处理人乳腺癌MDA-MB-231细胞及正常乳腺上皮细胞MCF-10A后,采用CCK-8方法检测不同浓度的DP(0、100、200、400、800 μg/mL)对细胞活力的影响;采用平板克隆实验检测DP对乳腺癌细胞克隆形成能力的影响;采用划痕实验和Transwell实验分别检测DP对乳腺癌迁移和侵袭能力的影响;蛋白免疫印迹法(Western blotting)检测DP作用MDA-MB-231细胞48 h后,该细胞中磷脂酰肌醇3激酶(PI3K)、磷酸化磷脂酰肌醇3激酶(p-PI3K)、蛋白激酶 B(Akt)、磷酸化蛋白激酶 B(p-Akt)、糖原合成激酶-3β(GSK-3β)、磷酸化糖原合成激酶-3β(p-GSK-3β)蛋白表达情况,上皮间质转化(EMT)相关标志蛋白E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)表达情况。结果显示,与空白组相比,DP组(100、200、400、800 μg/mL)能够显著抑制MDA-MB-231细胞存活率(P<0.05或P<0.01),而对MCF-10A细胞的存活率无显著影响;与空白组相比,DP组(200、400 μg/mL)细胞克隆形成能力显著减弱(P<0.01);与空白组相比,DP组(200、400 μg/mL)迁移能力显著降低;与空白组相比,DP组(200、400 μg/mL)侵袭能力显著减弱(P<0.01);与空白组相比,DP组(200、400 μg/mL)p-PI3K、p-Akt及p-GSK-3β蛋白表达水平降低(P<0.01),而PI3K、Akt及GSK-3β总蛋白无明显变化;与空白组相比,DP组(200、400 μg/mL)E-cadherin表达水平上调,N-cadherin和Vimentin表达水平下调,差异均有统计学意义(P<0.05或P<0.01)。综上,DP能够有效抑制人乳腺癌MDA-MB-231细胞增殖、迁移、侵袭和EMT进程,其机制可能与抑制PI3K/Akt/GSK-3β信号通路活性有关。

关键词: 磷脂酰肌醇3激酶/蛋白激酶B/糖原合成激酶-3β, 蒲公英多糖, 乳腺癌, 增殖, 迁移, 侵袭

Abstract:

In this study,we investigated the molecular mechanism of dandelion polysaccharide (DP) inhibition of breast cancer cells by observing the effects of DP on the proliferation,migration and invasion of human triple-negative breast cancer MDA-MB-231 cells.After treating human breast cancer MDA-MB-231 cells and normal breast epithelial cells MCF-10A with DP,the effect of different concentrations of DP (0,100,200,400,800 μg/mL) on cell viability was detected by CCK-8 method. The effect of DP on the clonogenic ability of breast cancer cells was detected by plate cloning assay.the effect of DP on the migration and invasion ability of breast cancer cells was examined by the scratch assay and Transwell assay.Western blotting was performed to detect phosphatidylinositol 3 kinase (PI3K),phosphorylated phosphatidylinositol 3 kinase (p-PI3K),protein kinase B (Akt),phosphorylated protein kinase B (p-Akt),glycogen synthase kinase-3β (GSK-3β),phosphorylated glycogen synthase kinase-3β (p-GSK-3β) protein expression,epithelial mesenchymal transition (EMT)-related marker proteins E-cadherin,N-cadherin and vimentin expression.The results showed that compared with the blank group,the DP group (100,200,400,800 μg/mL) was able to significantly inhibit the viability of MDA-MB-231 cells (< 0.05 or < 0.01),and there was no significant effect on the viability of MCF-10A cells.compared with the blank group,the DP group (200,400 μg/mL) was able to significantly reduce the ability of cell clone formation (< 0.01).the DP group (200,400 μg/mL) showed significantly reduced migration and invasion ability ( < 0.01).the DP group (200,400 μg/mL) showed significantly reduced p-PI3K,p-Akt and p-GSK-3β protein expression levels were decreased in the DP group (200,400 μg/mL) compared with the blank group (< 0.01),while no significant changes were observed in PI3K,Akt and GSK-3β total proteins.E-cadherin expression levels were upregulated,but N-cadherin and Vimentin expression levels were downregulated in the DP group (200,400 μg/mL) compared with the blank group,and the differences were statistically significant ( < 0.05 or P < 0.01).In conclusion,DP can effectively inhibit the proliferation,migration,invasion and EMT process of human breast cancer MDA-MB-231 cells,and the mechanism may be related to the inhibition of PI3K/Akt/GSK-3β signaling pathway activity.

Key words: PI3K/Akt/GSK-3β, dandelion polysaccharide, breast cancer, proliferation, migration, invasion

中图分类号:  R966