白头翁皂苷D通过多靶点和多途径抑制三阴性乳腺癌侵袭转移

doi:10.12122/j.issn.1673-4254.2025.01.18

摘要/Abstract

摘要：

目的采用网络药理学和计算机模拟探讨白头翁皂苷D（PSD）抑制三阴性乳腺癌（TNBC）侵袭转移的作用机制，并进行实验验证。方法运用Super-PRED、Swiss Target Prediction、PharmMapper、STITCH和BATMAN-TCM数据库收集PSD潜在靶点，利用GeneCards和OMIM数据库获得TNBC侵袭转移靶点，将两者相交获得药物-疾病交集靶点，利用Cytoscape3.10.1软件绘制“PSD-靶点-疾病”互作网络。运用Cytoscape3.10.1中的Centiscape2.2插件设定阈值并获得核心靶点，使用String数据库进行蛋白质互作（PPI）分析，通过David数据库对核心靶点进行KEGG通路和GO功能富集分析。最后将核心靶点依次与PSD进行分子对接。通过Transwell和Western blotting法对PSD的作用及机制进行验证。结果网络药理学结果显示，共筛选出PSD潜在靶点285个及药物与疾病核心靶点26个。GO分析获得175个条目，涉及生物大分子（蛋白质、DNA、RNA）的结合、酶活性、基因转录调控等方面。KEGG分析获得46个条目，涉及癌症途径、化学致癌-受体活化、癌症中的微小RNA、化学致癌-活性氧、癌症中PD-L1表达和PD-1检查点通路等。分子对接显示，PSD与MTOR、HDAC2、ABL1、CDK1、TLR4、TERT、PIK3R1、NFE2L2、PTPN1有较高的结合性。Transwell和Western blotting结果显示，PSD抑制TNBC细胞侵袭迁移且降低其MMP2、MMP9、N-cadherin及关键蛋白p-mTOR、ABL1、TERT、PTPN1、HDAC2、PIK3R1、CDK1、TLR4和细胞核内NFE2L2的表达（P<0.05），PSD可通过这些靶点阻碍TNBC侵袭转移。结论 PSD通过多靶点和多途径抑制TNBC侵袭转移，为后续深入的机制研究提供前期基础，为TNBC药物研发提供新思路。

关键词: 白头翁皂苷D, 网络药理学, 分子对接, 三阴性乳腺癌, 侵袭, 转移

Abstract:

Objective To explore the mechanism by which Pulsatilla saponin D (PSD) inhibits invasion and metastasis of triple-negative breast cancer (TNBC). Methods The public databases were used to identify the potential targets of PSD and the invasion and metastasis targets of TNBC to obtain the intersection targets between PSD and TNBC. The "PSD-target-disease" interaction network was constructed and protein-protein interaction (PPI) analysis was performed to obtain the core targets, which were analyzed for KEGG pathway and GO functional enrichment. Molecular docking study of the core targets and PSD was performed, and the therapeutic effect and mechanism of PSD were verified using Transwell assay and Western blotting in cultured TNBC cells. Results Network pharmacology analysis identified a total of 285 potential PSD targets and 26 drug-disease intersection core targets. GO analysis yielded 175 entries related to the binding of biomolecules (protein, DNA and RNA), enzyme activities, and regulation of gene transcription. KEGG analysis yielded 46 entries involving pathways in cancer, chemical carcinogenesis-receptor activation, microRNAs in cancer, chemical carcinogenesis-reactive oxygen species, PD-L1 expression and PD-1 checkpoint pathway in cancer. Molecular docking showed high binding affinities of PSD to MTOR, HDAC2, ABL1, CDK1, TLR4, TERT, PIK3R1, NFE2L2 and PTPN1. In cultured TNBC cells, treatment with PSD significantly inhibited cell invasion and migration and lowered the expressions of MMP2, MMP9, N-cadherin and the core proteins p-mTOR, ABL1, TERT, PTPN1, HDAC2, PIK3R1, CDK1, TLR4 as well as NFE2L2 expressionin the cell nuclei. Conclusion The inhibitory effects of PSD on TNBC invasion and metastasis are mediated by multiple targets and pathways.

Key words: Pulsatilla saponin D, network pharmacology, molecular docking, triple negative breast cancer, invasion, metastasis

褚乔, 王小娜, 续佳颖, 彭荟林, 赵裕琳, 张静, 陆国玉, 王恺. 白头翁皂苷D通过多靶点和多途径抑制三阴性乳腺癌侵袭转移[J]. 南方医科大学学报, 2025, 45(1): 150-161.

Qiao CHU, Xiaona WANG, Jiaying XU, Huilin PENG, Yulin ZHAO, Jing ZHANG, Guoyu LU, Kai WANG. Pulsatilla saponin D inhibits invasion and metastasis of triple-negative breast cancer cells through multiple targets and pathways[J]. Journal of Southern Medical University, 2025, 45(1): 150-161.

图/表 12

参考文献 37

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Core targets	Degree unDir	PDB ID
HSP90AA1	39	6GR5
HSP90AB1	33	6N8Y
NFKB1	27	1SVC
MTOR	24	4JSN
HDAC2	23	7JS8
ABL1	20	5HU9
CDK1	19	6GU2
TLR4	18	2Z62
TERT	17	5UGW
AR	17	8E1A
PIK3R1	17	7PG5
NFE2L2	16	7X5G
ABCB1	12	7A69
KDM1A	12	7E0G
CFTR	11	5TFJ
PTPN1	11	7MN9
PTGS1	10	6Y3C
GRIA2	9	7F3O
MME	8	6SUK
PTK2B	7	3CC6
ACACA	6	2YL2
FPR2	6	6LW5
CAPN1	6	7W7O
SLC1A2	5	7VR7
LTA4H	4	3U9W
MMP1	4	966C

Core targets	Degree unDir	PDB ID
HSP90AA1	39	6GR5
HSP90AB1	33	6N8Y
NFKB1	27	1SVC
MTOR	24	4JSN
HDAC2	23	7JS8
ABL1	20	5HU9
CDK1	19	6GU2
TLR4	18	2Z62
TERT	17	5UGW
AR	17	8E1A
PIK3R1	17	7PG5
NFE2L2	16	7X5G
ABCB1	12	7A69
KDM1A	12	7E0G
CFTR	11	5TFJ
PTPN1	11	7MN9
PTGS1	10	6Y3C
GRIA2	9	7F3O
MME	8	6SUK
PTK2B	7	3CC6
ACACA	6	2YL2
FPR2	6	6LW5
CAPN1	6	7W7O
SLC1A2	5	7VR7
LTA4H	4	3U9W
MMP1	4	966C

ID	Pathways	P	Number of related genes
hsa05200	Pathways in cancer	0.000000974	11
hsa05417	Lipid and atherosclerosis	0.000026000	7
hsa05215	Prostate cancer	0.000007060	6
hsa04613	Neutrophil extracellular trap formation	0.000184000	6
hsa05170	Human immunodeficiency virus 1 infection	0.000298000	6
hsa05207	Chemical carcinogenesis - receptor activation	0.000298000	6
hsa05206	MicroRNAs in cancer	0.001673574	6
hsa04151	PI3K-Akt signaling pathway	0.003182088	6
hsa05418	Fluid shear stress and atherosclerosis	0.000618000	5
hsa04218	Cellular senescence	0.000953000	5
hsa05208	Chemical carcinogenesis-reactive oxygen species	0.003535389	5
hsa05131	Shigellosis	0.005091764	5
hsa05165	Human papillomavirus infection	0.014031422	5
hsa05010	Alzheimer disease	0.022984313	5
hsa05220	Chronic myeloid leukemia	0.001299397	4
hsa05235	PD-L1 expression and PD-1 checkpoint pathway in cancer	0.002047819	4
hsa04657	IL-17 signaling pathway	0.002394257	4
hsa04914	Progesterone-mediated oocyte maturation	0.003020588	4
hsa04931	Insulin resistance	0.003550859	4
hsa04659	Th17 cell differentiation	0.003550859	4
hsa04066	HIF-1 signaling pathway	0.003644426	4
hsa04152	AMPK signaling pathway	0.004886673	4
hsa04910	Insulin signaling pathway	0.006900929	4
hsa05135	Yersinia infection	0.006900929	4
hsa05226	Gastric cancer	0.008693554	4
hsa04217	Necroptosis	0.010557294	4
hsa05161	Hepatitis B	0.010919274	4
hsa05225	Hepatocellular carcinoma	0.012048446	4
hsa04141	Protein processing in endoplasmic reticulum	0.012439336	4
hsa04621	NOD-like receptor signaling pathway	0.015830764	4
hsa05203	Viral carcinogenesis	0.020215918	4
hsa05163	Human cytomegalovirus infection	0.026104939	4
hsa04024	cAMP signaling pathway	0.026104939	4
hsa05171	Coronavirus disease - COVID-19	0.028568364	4
hsa05132	Salmonella infection	0.033858771	4
hsa05016	Huntington disease	0.056576135	4
hsa04213	Longevity regulating pathway - multiple species	0.013188318	3
hsa05221	Acute myeloid leukemia	0.015766996	3
hsa05212	Pancreatic cancer	0.020005321	3
hsa04012	ErbB signaling pathway	0.02466712	3
hsa04211	Longevity regulating pathway	0.026868984	3
hsa05146	Amoebiasis	0.034548353	3
hsa05142	Chagas disease	0.034548353	3
hsa04620	Toll-like receptor signaling pathway	0.038349814	3
hsa04722	Neurotrophin signaling pathway	0.045714487	3
hsa04919	Thyroid hormone signaling pathway	0.047106228	3

ID	Pathways	P	Number of related genes
hsa05200	Pathways in cancer	0.000000974	11
hsa05417	Lipid and atherosclerosis	0.000026000	7
hsa05215	Prostate cancer	0.000007060	6
hsa04613	Neutrophil extracellular trap formation	0.000184000	6
hsa05170	Human immunodeficiency virus 1 infection	0.000298000	6
hsa05207	Chemical carcinogenesis - receptor activation	0.000298000	6
hsa05206	MicroRNAs in cancer	0.001673574	6
hsa04151	PI3K-Akt signaling pathway	0.003182088	6
hsa05418	Fluid shear stress and atherosclerosis	0.000618000	5
hsa04218	Cellular senescence	0.000953000	5
hsa05208	Chemical carcinogenesis-reactive oxygen species	0.003535389	5
hsa05131	Shigellosis	0.005091764	5
hsa05165	Human papillomavirus infection	0.014031422	5
hsa05010	Alzheimer disease	0.022984313	5
hsa05220	Chronic myeloid leukemia	0.001299397	4
hsa05235	PD-L1 expression and PD-1 checkpoint pathway in cancer	0.002047819	4
hsa04657	IL-17 signaling pathway	0.002394257	4
hsa04914	Progesterone-mediated oocyte maturation	0.003020588	4
hsa04931	Insulin resistance	0.003550859	4
hsa04659	Th17 cell differentiation	0.003550859	4
hsa04066	HIF-1 signaling pathway	0.003644426	4
hsa04152	AMPK signaling pathway	0.004886673	4
hsa04910	Insulin signaling pathway	0.006900929	4
hsa05135	Yersinia infection	0.006900929	4
hsa05226	Gastric cancer	0.008693554	4
hsa04217	Necroptosis	0.010557294	4
hsa05161	Hepatitis B	0.010919274	4
hsa05225	Hepatocellular carcinoma	0.012048446	4
hsa04141	Protein processing in endoplasmic reticulum	0.012439336	4
hsa04621	NOD-like receptor signaling pathway	0.015830764	4
hsa05203	Viral carcinogenesis	0.020215918	4
hsa05163	Human cytomegalovirus infection	0.026104939	4
hsa04024	cAMP signaling pathway	0.026104939	4
hsa05171	Coronavirus disease - COVID-19	0.028568364	4
hsa05132	Salmonella infection	0.033858771	4
hsa05016	Huntington disease	0.056576135	4
hsa04213	Longevity regulating pathway - multiple species	0.013188318	3
hsa05221	Acute myeloid leukemia	0.015766996	3
hsa05212	Pancreatic cancer	0.020005321	3
hsa04012	ErbB signaling pathway	0.02466712	3
hsa04211	Longevity regulating pathway	0.026868984	3
hsa05146	Amoebiasis	0.034548353	3
hsa05142	Chagas disease	0.034548353	3
hsa04620	Toll-like receptor signaling pathway	0.038349814	3
hsa04722	Neurotrophin signaling pathway	0.045714487	3
hsa04919	Thyroid hormone signaling pathway	0.047106228	3

Compound	Targets	PDB ID	MM-GBSA dG Bind (kcal/mol)
PSD	MTOR	4JSN	-32.00
	HDAC2	7JS8	-17.49
	ABL1	5HU9	-30.56
	CDK1	6GU2	-32.77
	TLR4	2Z62	-19.87
	TERT	5UGW	-39.09
	PIK3R1	7PG5	-31.43
	NFE2L2	7X5G	-43.75
	KDM1A	7E0G	-43.68
	PTPN1	7MN9	-20.59