PMID- 38403323
OWN - NLM
STAT- MEDLINE
DCOM- 20240227
LR  - 20240227
IS  - 1001-5302 (Print)
IS  - 1001-5302 (Linking)
VI  - 49
IP  - 2
DP  - 2024 Jan
TI  - [Mechanism of n-butanol fraction of Wenxia Formula combining with gefitinib in 
      treating non-small cell lung cancer based on network pharmacology and in vitro 
      experiment].
PG  - 471-486
LID - 10.19540/j.cnki.cjcmm.20230914.704 [doi]
AB  - This study combined network pharmacology, molecular docking, and in vitro 
      experiments to explore the potential mechanism of the active components of the 
      n-butanol fraction of Wenxia Formula(NWXF) combined with gefitinib(GEF) in 
      treating non-small cell lung cancer(NSCLC). Ultra-performance liquid 
      chromatography-quadrupole Orbitrap mass spectrometry(UPLC-Q-Orbitrap MS) was 
      employed to detect the main chemical components of NWXF. The active components of 
      NWXF were retrieved from SwissADME, and the candidate targets of these active 
      components were retrieved from SwissTargetPrediction. Online Mendelian 
      Inheritance in Man(OMIM) and GeneCards were searched for the targets of NSCLC. 
      Cytoscape 3.9.0 and STRING were employed to build the protein-protein 
      interaction(PPI) network with the common targets shared by NWXF and NSCLC. Gene 
      Ontology(GO) annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) 
      enrichment were performed in DAVID to predict the potential mechanisms. Finally, 
      molecular docking between the main active ingredients and key targets was 
      conducted in SYBYL-X 2.0. The methyl thiazolyl tetrazolium(MTT) assay was 
      employed to evaluate the inhibitory effects of NWXF and/or GEF on the 
      proliferation of human non-small cell lung cancer cells(A549 and PC-9). 
      Additionally, the impact of NWXF on human embryonic lung fibroblast cells(MRC-5) 
      was assessed. The effectiveness of the drug combination was evaluated based on 
      the Q value. The terminal-deoxynucleoitidyl transferase mediated nick-end 
      labeling(TUNEL) assay was employed to examine the apoptosis of A549 and PC-9 
      cells treated with NWXF and/or GEF. Quantitative real-time PCR(qRT-PCR) was 
      employed to measure the mRNA levels of epidermal growth factor receptor(EGFR), 
      c-Jun N-terminal kinase(JNK), and Bcl2-associated X protein(Bax) in the A549 and 
      PC-9 cells treated with NWXF and/or GEF. Western blot was employed to determine 
      the protein levels of EGFR, p-EGFR, JNK, p-JNK, and Bax in the A549 and PC-9 
      cells treated with NWXF and/or GEF. A total of 77 active components, 488 
      potential targets, and 49 key targets involved in the treatment of NSCLC with 
      NWXF were predicted. The results of GO annotation showed that NWXF may treat 
      NSCLC by regulating the biological processes such as cell proliferation, 
      apoptosis, and protein phosphorylation. KEGG enrichment revealed that the key 
      targets of NWXF in treating NSCLC were enriched in the mitogen-activated protein 
      kinase(MAPK), phosphatidylinositol 3-kinase(PI3K)-protein kinase B(AKT), 
      hypoxia-inducible factor-1(HIF-1), and microRNA-related signaling pathways. 
      Molecular docking results showed that 91.9% of the docking scores were greater 
      than 5, indicating the strong binding capability between main active components 
      and key targets. The cell experiments demonstrated that NWXF combined with GEF 
      synergistically inhibited the proliferation, promoted the apoptosis, decreased 
      p-EGFR/EGFR and p-JNK/JNK values, down-regulated the mRNA levels of EGFR and JNK, 
      and up-regulated the mRNA and protein levels of Bax in A549 and PC-9 cells. In 
      conclusion, NWXF combined with GEF can regulate the EGFR/JNK pathway to promote 
      the apoptosis of NSCLC cells, thus treating NSCLC.
FAU - Chen, Rui-Jie
AU  - Chen RJ
AD  - Huzhou Central Hospital Huzhou 313000, China.
FAU - Bi, Qian-Yu
AU  - Bi QY
AD  - School of Basic Medicine, Zhejiang Chinese Medical University Hangzhou 310053, 
      China.
FAU - Shi, Jie-Min
AU  - Shi JM
AD  - Huzhou Central Hospital Huzhou 313000, China.
FAU - Zhong, Jing
AU  - Zhong J
AD  - Huzhou Central Hospital Huzhou 313000, China.
FAU - Han, Jin-Tao
AU  - Han JT
AD  - Huzhou Central Hospital Huzhou 313000, China.
FAU - Ji, Xu-Ming
AU  - Ji XM
AD  - Laboratory of Neuropharmacology and Translational Medicine of Zhejiang Province 
      Hangzhou 310053, China Academy of Chinese Medical Sciences, Zhejiang Chinese 
      Medical University Hangzhou 310053, China.
LA  - chi
PT  - English Abstract
PT  - Journal Article
PL  - China
TA  - Zhongguo Zhong Yao Za Zhi
JT  - Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese 
      materia medica
JID - 8913656
RN  - S65743JHBS (Gefitinib)
RN  - 8PJ61P6TS3 (1-Butanol)
RN  - 0 (bcl-2-Associated X Protein)
RN  - EC 2.7.1.- (Phosphatidylinositol 3-Kinases)
RN  - EC 2.7.10.1 (ErbB Receptors)
RN  - 0 (RNA, Messenger)
RN  - 0 (Drugs, Chinese Herbal)
SB  - IM
MH  - Humans
MH  - *Carcinoma, Non-Small-Cell Lung/drug therapy/genetics
MH  - Gefitinib/pharmacology
MH  - 1-Butanol
MH  - bcl-2-Associated X Protein
MH  - Network Pharmacology
MH  - Molecular Docking Simulation
MH  - Phosphatidylinositol 3-Kinases
MH  - *Lung Neoplasms/drug therapy/genetics
MH  - ErbB Receptors
MH  - RNA, Messenger
MH  - *Drugs, Chinese Herbal/pharmacology
OTO - NOTNLM
OT  - apoptosis
OT  - gefitinib
OT  - mechanism
OT  - mole-cular docking
OT  - n-butanol fraction of Wenxia Formula
OT  - network pharmacology
OT  - non-small cell lung cancer
EDAT- 2024/02/26 00:42
MHDA- 2024/02/27 06:44
CRDT- 2024/02/25 20:52
PHST- 2024/02/27 06:44 [medline]
PHST- 2024/02/26 00:42 [pubmed]
PHST- 2024/02/25 20:52 [entrez]
AID - 10.19540/j.cnki.cjcmm.20230914.704 [doi]
PST - ppublish
SO  - Zhongguo Zhong Yao Za Zhi. 2024 Jan;49(2):471-486. doi: 
      10.19540/j.cnki.cjcmm.20230914.704.