Mass-spectrometric and bioinformatic analysis of eEF1Bγ interactome in the cytoplasmic fraction of A549 cells

Abstract Purpose: There is an urgent need to develop effective anti-pneumonia drugs. Phillygenin (PHI) is derived from Forsythia suspense and possesses anti-inflammation, anti-oxidant bioactivities. In the present study, we aimed to evaluate the therapeutic potential of phillygenin (PHI) on lipopolysaccharide (LPS)-induced acute pneumonia.Methods: The molecular target of PHI was predicted by bioinformatic analysis. Hollow fiber-based ligand fishing (HFLF) strategy and luciferase reporter assay were further used to identify the target of PHI. LPS-induced acute pneumonia rat model and A549 cells model were used to evaluate PHI function. TNF-α pathway and apoptosis associated proteins were detected by Western Blot, immunofluorescence and immunohistochemistry. Cell cycle and cytokines were determined by flow cytometry.Results: The bioinformatic analysis and luciferase reporter assay identified that the target protein of PHI was pregnane X receptor (PXR) PHI could directly bind to PXR protein and inhibit NF-κB P65 activity. PHI significantly decreased the expression of phosphorylated JNK, P38, Erk, P65 in acute pneumonia rat model. PHI also declined the expression of Bax, Caspase-3 and Caspase-9 and repressed lung epithelial cell apoptosis induced by LPS in vivo and in vitro. In addition, PHI inhibited inflammation cytokines production including TNF-α, IFN-γ, IL-6, IL-1β and IL-18.Conclusions: PHI significantly alleviated LPS-induced lung injury in vivo by exerting anti-inflammatory effects. This is the first study to demonstrate that PHI, a small molecule natural product, significantly alleviates LPS-induced acute pneumonia by binding to PXR. Thus, PHI can be a novel therapeutic agent for pneumonia.

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Crystal Structures and Cytotoxicity of ent-Kaurane-Type Diterpenoids from Two Aspilia Species

Molecules ◽

10.3390/molecules23123199 ◽

2018 ◽

Vol 23 (12) ◽

pp. 3199 ◽

Cited By ~ 3

Author(s):

Souaibou Yaouba ◽

Arto Valkonen ◽

Paolo Coghi ◽

Jiaying Gao ◽

Eric M. Guantai ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Nuclear Magnetic Resonance ◽

Nmr Assignments ◽

A549 Cells ◽

Mass Spectrometric ◽

X Ray Diffraction ◽

Hep G2 ◽

X Ray ◽

Aerial Parts ◽

Phytochemical Investigation

A phytochemical investigation of the roots of Aspilia plurisetaled to the isolation of ent-kaurane-type diterpenoids and additional phytochemicals (1–23). The structures of the isolated compounds were elucidated based on Nuclear Magnetic Resonance (NMR) spectroscopic and mass spectrometric analyses. The absolute configurations of seven of the ent-kaurane-type diterpenoids (3–6, 6b, 7 and 8) were determined by single crystal X-ray diffraction studies. Eleven of the compounds were also isolated from the roots and the aerial parts of Aspilia mossambicensis. The literature NMR assignments for compounds 1 and 5 were revised. In a cytotoxicity assay, 12α-methoxy-ent-kaur-9(11),16-dien-19-oic acid (1) (IC50 = 27.3 ± 1.9 µM) and 9β-hydroxy-15α-angeloyloxy-ent-kaur-16-en-19-oic acid (3) (IC50 = 24.7 ± 2.8 µM) were the most cytotoxic against the hepatocellular carcinoma (Hep-G2) cell line, while 15α-angeloyloxy-16β,17-epoxy-ent-kauran-19-oic acid (5) (IC50 = 30.7 ± 1.7 µM) was the most cytotoxic against adenocarcinomic human alveolar basal epithelial (A549) cells.

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MiR-199a-5p-HIF-1α-STAT3 positive feed-back loop contributes to the progression of NSCLC

10.21203/rs.3.rs-52959/v1 ◽

2020 ◽

Author(s):

Xingping Yang ◽

Yuzhen Zheng ◽

Jian Tan ◽

Xiansheng Hu ◽

Renjiang Tian ◽

...

Keyword(s):

A549 Cells ◽

Bioinformatic Analysis ◽

Luciferase Assay ◽

Migration And Invasion ◽

Flow Cytometry Analysis ◽

Transwell Assay ◽

Xenograft Models ◽

Direct Target ◽

Induced Apoptosis ◽

Common Malignancy

Abstract Background Non-small cell lung cancer (NSCLC), is the most common malignancy worldwide. MiR-199a-5p has been reported to play important roles in multiple tumors, inclusive of NSCLC. Whereas, little is definitively known pertaining to its explicit mechanism of action in NSCLC. Methods Expression of miR-199a-5p and HIF-1α mRNA were quantified employing qRT-PCR. H1299 and A549 cells were transiently transfected with miR-199a-5p mimics or inhibitors. Then CCK-8 assays, flow cytometry analysis, Transwell assay were implemented for detecting cell proliferation, cell cycle, apoptosis, migration and invasion of NSCLC cells, respectively. HIF-1α, STAT3 and p-STAT3 expression were detected via Western blotting. Bioinformatic analysis and dual-luciferase assay were performed for monitoring interaction between miR-199a-5p, HIF-1α and STAT3. Xenograft models were established with nude mice for further analyzing Bevacizumab resistance of NSCLC. Results MiR-199a-5p expression was markedly attenuated in NSCLC tissues and cell lines. Overexpression of miR-199a-5p constrained proliferation, migration, invasion but induced apoptosis of NSCLC cells. HIF-1α was identified as a direct target of miR-199a-5p. Further study confirmed a positive feed-back loop between miR-199a-5p, HIF-1α and STAT3. Co-transfection of HIF-1α or STAT3 overexpression plasmids counteracted effects of miR-199a-5p. Further study substantiated that feed-back loop might be in association with the Bevacizumab resistance of NSCLC cells . Conclusion MiR-199a-5p constrained the progression and Bevacizumab resistance of NSCLC by suppressing HIF-1α and STAT3, while HIF-1α/STAT3 axis suppressed the expression of miR-199a-5p, which forms a positive feed-back loop to promote the sustaining progression of NSCLC.

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Bioinformatic analysis of miR-4792 regulates Radix Tetrastigma hemsleyani flavone to inhibit proliferation, invasion, and induce apoptosis of A549 cells

OncoTargets and Therapy ◽

10.2147/ott.s182525 ◽

2019 ◽

Vol Volume 12 ◽

pp. 1401-1412 ◽

Cited By ~ 3

Author(s):

Peigang Liu ◽

Jinbao Pu ◽

Junhui Zhang ◽

Zhilu Chen ◽

Kemin Wei ◽

...

Keyword(s):

A549 Cells ◽

Bioinformatic Analysis

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Advanced bioinformatic analysis and pathway prediction of NSCLC cells upon cisplatin resistance

Scientific Reports ◽

10.1038/s41598-021-85930-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

A K M Nawshad Hossian ◽

Fatema Tuz Zahra ◽

Sagun Poudel ◽

Camille F. Abshire ◽

Paula Polk ◽

...

Keyword(s):

Cholesterol Biosynthesis ◽

A549 Cells ◽

Bioinformatic Analysis ◽

Rna Seq ◽

Prolonged Incubation ◽

Pathway Activity ◽

Gene Dysregulation ◽

Pathway Prediction ◽

Bioinformatics Software ◽

A549 Lung

AbstractThis study aims to identify pathway involvement in the development of cisplatin (cis-diamminedichloroplatinum (II); CDDP) resistance in A549 lung cancer (LC) cells by utilizing advanced bioinformatics software. We developed CDDP-resistant A549 (A549/DDP) cells through prolonged incubation with the drug and performed RNA-seq on RNA extracts to determine differential mRNA and miRNA expression between A549/DDP and A549 cells. We analyzed the gene dysregulation with Ingenuity Pathway Analysis (IPA; QIAGEN) software. In contrast to prior research, which relied on the clustering of dysregulated genes to pathways as an indication of pathway activity, we utilized the IPA software for the dynamic evaluation of pathway activity depending on the gene dysregulation levels. We predicted 15 pathways significantly contributing to the chemoresistance, with several of them to have not been previously reported or analyzed in detail. Among them, the PKR signaling, cholesterol biosynthesis, and TEC signaling pathways are included, as well as genes, such as PIK3R3, miR-34c-5p, and MDM2, among others. We also provide a preliminary analysis of SNPs and indels, present exclusively in A549/DDP cells. This study's results provide novel potential mechanisms and molecular targets that can be explored in future studies and assist in improving the understanding of the chemoresistance phenotype.

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