Mass Spectrometry-Based Proteomics Approach Characterizes the Dual Functionality of miR-328 in Monocytes

Saul, Meike J.; Hegewald, Anett B.; Emmerich, Anne C.; Ossipova, Elena; Vogel, Marc; Baumann, Isabell; Kultima, Kim; Lengqivst, Johan; Steinhilber, Dieter; Jakobsson, Per Johan

doi:10.3389/fphar.2019.00640

Cited by 6 publications

(7 citation statements)

References 17 publications

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“…A FC cutoff of ≥1.25 for overexpression and ≤0.8 for the downregulation of proteins was selected to be biologically significant, and an adjusted p -value, that is, q ≤ 0.05, was selected for statistical significance. A similar range of FCs has been previously reported by studies using isobaric labeling. − Rotenone treatment highlighted 421 differentially regulated proteins (FC ≥1.25 or ≤0.8), of which 144 were significantly altered ( q ≤ 0.05, with respect to untreated cell control, Figure B). Yashtimadhu choorna extract cotreatment was found to significantly regulate 88 proteins ( q ≤ 0.05, with respect to rotenone), which included restoration of 84 proteins that were dysregulated by rotenone and an additional regulation of 4 proteins (Figure C).…”

Section: Results and Discussionsupporting

confidence: 82%

Identification of Molecular Network Associated with Neuroprotective Effects of Yashtimadhu (Glycyrrhiza glabra L.) by Quantitative Proteomics of Rotenone-Induced Parkinson’s Disease Model

et al. 2020

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Parkinson’s disease (PD) is a progressive neurodegenerative disorder, whose treatment with modern therapeutics leads to a plethora of side effects with prolonged usage. Therefore, the management of PD with complementary and alternative medicine is often pursued. In the Ayurveda system of alternative medicine, Yashtimadhu choorna, a Medhya Rasayana (nootropic), prepared from the dried roots of Glycyrrhiza glabra L. (licorice), is prescribed for the management of PD with a favorable outcome. We pursued to understand the neuroprotective effects of Yashtimadhu choorna against a rotenone-induced cellular model of PD using differentiated IMR-32 cells. Cotreatment with Yashtimadhu choorna extract rescued rotenone-induced apoptosis and hyperphosphorylation of ERK-1/2. Quantitative proteomic analysis of six peptide fractions from independent biological replicates acquired 1,561,169 mass spectra, which when searched resulted in 565,008 peptide-spectrum matches mapping to 30,554 unique peptides that belonged to 4864 human proteins. Proteins commonly identified in biological replicates and >4 PSMs were considered for further analysis, leading to a refined set of 3720 proteins. Rotenone treatment differentially altered 144 proteins (fold ≥1.25 or ≤0.8), involved in mitochondrial, endoplasmic reticulum, and autophagy functions. Cotreatment with Yashtimadhu choorna extract rescued 84 proteins from the effect of rotenone and an additional regulation of 4 proteins. Network analysis highlighted the interaction of proteins and pathways regulated by them, which can be targeted for neuroprotection. Validation of proteomics data highlighted that Yashtimadhu confers neuroprotection by preventing mitochondrial oxidative stress and apoptosis. This discovery will pave the way for understanding the molecular action of Ayurveda drugs and developing novel therapeutics for PD.

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Section: Results and Discussionsupporting

confidence: 82%

Identification of Molecular Network Associated with Neuroprotective Effects of Yashtimadhu (Glycyrrhiza glabra L.) by Quantitative Proteomics of Rotenone-Induced Parkinson’s Disease Model

et al. 2020

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“…Interestingly, the decoy mechanism of miR-328 and hnRNP E2 affects a variety of targets and cellular processes (Saul et al, 2016(Saul et al, , 2019b while in this case the decoy of miR-574-5p and CUGBP1 seems to be quite specific for mPGES-1. Retrospectively, this is consistent with one of our previous studies in which we firstly demonstrated that miR-574-5p prevents CUGBP1 from binding to mPGES-1 mRNA.…”

Section: Discussionmentioning

confidence: 99%

“…However, it should be mentioned that mPGES-1 did not reveal this pattern in the proteomics data. We would like to point out that a proteomics study, even if we have used an established method (Eriksson et al, 2008;Ochs et al, 2013;Bergqvist et al, 2019;Saul et al, 2019b), only provides initial indications of which proteins could be regulated. It does not map the exact expression changes, which is a common limitation of this approach.…”

Section: Discussionmentioning

confidence: 99%

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Proteomics-Based Characterization of miR-574-5p Decoy to CUGBP1 Suggests Specificity for mPGES-1 Regulation in Human Lung Cancer Cells

et al. 2020

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MicroRNAs (miRs) are one of the most important post-transcriptional repressors of gene expression. However, miR-574-5p has recently been shown to positively regulate the expression of microsomal prostaglandin E-synthase-1 (mPGES-1), a key enzyme in the prostaglandin E 2 (PGE 2 ) biosynthesis, by acting as decoy to the RNA-binding protein CUG-RNA binding protein 1 (CUGBP1) in human lung cancer. miR-574-5p exhibits oncogenic properties and promotes lung tumor growth in vivo via induction of mPGES-1-derived PGE 2 synthesis. In a mass spectrometry-based proteomics study, we now attempted to characterize this decoy mechanism in A549 lung cancer cells at a cellular level. Besides the identification of novel CUGBP1 targets, we identified that the interaction between miR-574-5p and CUGBP1 specifically regulates mPGES-1 expression. This is supported by the fact that CUGBP1 and miR-574-5p are located in the nucleus, where CUGBP1 regulates alternative splicing. Further, in a bioinformatical approach we showed that the decoy-dependent mPGES-1 splicing pattern is unique. The specificity of miR-574-5p/CUGBP1 regulation on mPGES-1 expression supports the therapeutic strategy of pharmacological inhibition of PGE 2 formation, which may provide significant therapeutic value for NSCLC patients with high miR-574-5p levels.

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“…MiR-328 has been reported to function as a cancer-related miRNA. MiR-328 could increase reactive oxygen species (ROS) production and promote the adhesion and migration ability of monocytes during monocyte maturation (13). In lung cancer, miR-328 expression was significantly elevated and could promote cell migration (14)(15)(16).…”

Section: Introductionmentioning

confidence: 99%

Overexpression of miR-328-5p influences cell growth and migration to promote NSCLC progression by targeting LOXL4

Ji¹,

You²,

Wu³

et al. 2022

Ann Transl Med

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Background: Lung cancer is the leading cause of cancer-associated mortality worldwide, and most lung cancers are classified as non-small cell lung cancer (NSCLC). MiR-328 influence the progression of multiple tumors, but the role of miR-328-5p in NSCLC has not been elucidated. The aim of this study was to illuminate the oncogenic role and potential molecular mechanisms of the miR-328-5p and lysyl oxidase like 4 (LOXL4) in NSCLC.Methods: Expression of miR-328-5p was detected by real-time quantitative polymerase chain reaction (qRT-PCR) in tumor and non-tumor adjacent tissues. After Lentivirus-miR-328-5p was employed to intervene this miRNA in NSCLC cell lines, RT-qPCR was used to detect the expression levels of miR-328-5p. Cell Counting Kit-8 (CCK-8), cell colony formation, flow cytometry, wound healing, Transwell assays were used to determine the malignant phenotypes of NSCLC cells. Nude mice models of subcutaneous tumors were established to observe the effect of miR-328-5p on tumorigenesis. Targeting the 3'UTR of LOXL4 by miR-328-5p was verified by integrated analysis including transcriptome sequencing, dualluciferase and western-blot assays.Results: High miR-328-5p level was observed in NSCLC cells from The Cancer Genome Atlas (TCGA) database and tumor tissues collected from NSCLC patients. Overexpressed miR-328-5p promoted NSCLC cell proliferation, survival, and migration, and promoted tumor growth in vivo. Knockdown of miR-328-5p suppressed tumorigenic activities. Transcriptome sequencing analysis revealed that LOXL4 was downregulated by miR-328-5p, which was confirmed by dual-luciferase reporter and western-blot assays.Conclusions: miR-328-5p showed targeted regulation of LOXL4 to promote cell proliferation and migration in NSCLC.

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Mass Spectrometry-Based Proteomics Approach Characterizes the Dual Functionality of miR-328 in Monocytes

Cited by 6 publications

References 17 publications

Identification of Molecular Network Associated with Neuroprotective Effects of Yashtimadhu (Glycyrrhiza glabra L.) by Quantitative Proteomics of Rotenone-Induced Parkinson’s Disease Model

Identification of Molecular Network Associated with Neuroprotective Effects of Yashtimadhu (Glycyrrhiza glabra L.) by Quantitative Proteomics of Rotenone-Induced Parkinson’s Disease Model

Proteomics-Based Characterization of miR-574-5p Decoy to CUGBP1 Suggests Specificity for mPGES-1 Regulation in Human Lung Cancer Cells

Overexpression of miR-328-5p influences cell growth and migration to promote NSCLC progression by targeting LOXL4

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