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毛蕊异黄酮诱导人甲状腺乳头状癌细胞自噬和凋亡以及Sestrin2/AMPK/mTOR通路。

Calycosin induces autophagy and apoptosis Sestrin2/AMPK/mTOR in human papillary thyroid cancer cells.

作者信息

Qu Na, Qu Junsheng, Huang Na, Zhang Kexin, Ye Tongtong, Shi Junfeng, Chen Bing, Kan Chengxia, Zhang Jingwen, Han Fang, Hou Ningning, Sun Xiaodong, Pan Ruiyan

机构信息

Department of Endocrinology and Metabolism, Affiliated Hospital of Weifang Medical University, Weifang, China.

Clinical Research Center, Affiliated Hospital of Weifang Medical University, Weifang, China.

出版信息

Front Pharmacol. 2022 Dec 16;13:1056687. doi: 10.3389/fphar.2022.1056687. eCollection 2022.

DOI:10.3389/fphar.2022.1056687
PMID:36588732
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9800829/
Abstract

Calycosin, one of small molecules derived from astragalus, has anti-tumor effects in various tumors. However, the effects of calycosin on papillary thyroid cancer (PTC) remain unclear. This study aimed to explore the anti-tumor ability of calycosin on human PTC and its potential mechanisms. The B-CPAP cells were treated with calycosin, then cell proliferation, apoptosis and invasiveness were measured by CCK8 assay, flow cytometry, wound healing and transwell invasion assay, respectively. The cells were also performed by whole transcriptome microarray bioinformatics analysis. Apoptosis and autophagy-related markers or proteins were measured by qRT-PCR or western blot. Sestrin2-mediated AMPK/mTOR pathways were determined by western blot. We found that calycosin inhibited migration and invasion of B-CPAP cells and induced apoptosis (Bax/Bcl-2) and autophagy (LC3II/I, Beclin1) of B-CPAP cells. Differential expressed genes were screened between the calycosin-treated cells and control (524 genes upregulated and 328 genes downregulated). The pathway enrichment suggested that the role of calycosin in B-CPAP cells is closely related to apoptosis-related genes and p70S6 Kinase. Transmission electron microscopy found an increase in autophagosomes in calycosin-treated cells. Sestrin2 in human PTC tissues and B-CPAP cells was lower than in normal thyroid tissues and cells. And the pharmacological effects of calycosin in PTC cells were related to Sestrin2 activation, increased p-AMPK and inhibited p-mTOR and p-p70S6Kinase; these alterations were reversed when silencing Sestrin2. In conclusion, calycosin has an inhibitory effect on PTC promoting apoptosis and autophagy through the Sestrin2/AMPK/mTOR pathway.

摘要

毛蕊异黄酮是黄芪中的小分子成分之一，对多种肿瘤具有抗肿瘤作用。然而，毛蕊异黄酮对甲状腺乳头状癌（PTC）的作用尚不清楚。本研究旨在探讨毛蕊异黄酮对人PTC的抗肿瘤能力及其潜在机制。用毛蕊异黄酮处理B-CPAP细胞，然后分别通过CCK8法、流式细胞术、伤口愈合实验和Transwell侵袭实验检测细胞增殖、凋亡和侵袭能力。还对细胞进行了全转录组微阵列生物信息学分析。通过qRT-PCR或蛋白质免疫印迹法检测凋亡和自噬相关标志物或蛋白。通过蛋白质免疫印迹法确定Sestrin2介导的AMPK/mTOR信号通路。我们发现毛蕊异黄酮抑制B-CPAP细胞的迁移和侵袭，并诱导B-CPAP细胞凋亡（Bax/Bcl-2）和自噬（LC3II/I、Beclin1）。筛选出毛蕊异黄酮处理组细胞与对照组之间的差异表达基因（上调524个基因，下调328个基因）。通路富集分析表明，毛蕊异黄酮在B-CPAP细胞中的作用与凋亡相关基因和p70S6激酶密切相关。透射电子显微镜观察发现，毛蕊异黄酮处理组细胞中自噬体增多。人PTC组织和B-CPAP细胞中的Sestrin2低于正常甲状腺组织和细胞。毛蕊异黄酮在PTC细胞中的药理作用与Sestrin2激活、p-AMPK增加以及p-mTOR和p-p70S6激酶抑制有关；沉默Sestrin2后这些改变被逆转。总之，毛蕊异黄酮对PTC具有抑制作用，通过Sestrin2/AMPK/mTOR信号通路促进细胞凋亡和自噬。

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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/651a/9800829/325e18f884ee/fphar-13-1056687-g007.jpg
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