Quantitative proteomics and biochemical analyses reveal the role of endoplasmin in the regulation of the expression and secretion of A Disintegrin And Metalloproteinase 12

Xiong, Lipeng; Yan, Wenwen; Zubia, Emmanuel; Zhou, Yanqing; Zhang, Yang; Narayan, Mahesh; Xu, Guoqiang

doi:10.1016/j.jprot.2018.04.033

Cited by 4 publications

(7 citation statements)

References 54 publications

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“…Whether it means these patients have more active anti-tumor stromal cells and what types of stromal cells play a major anti-tumor role in these patients’ TME remain to be further studied. Among the three signature genes, ADAM12 was overexpressed in SCLC and might serve as a potential prognostic biomarker of SCLC (Duan et al, 2019; Shao et al, 2014; Xiong et al, 2018). Current study reveals that p65BTK, a novel isoform of the BTK, is overexpressed in NSCLC, and may be a novel drug target (Giordano et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

Identification of prognostic gene signature associated with microenvironment of lung adenocarcinoma

Cheng¹,

Ma²,

Zhou³

2019

PeerJ

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BackgroundLung cancer has the highest morbidity and mortality worldwide, and lung adenocarcinoma (LADC) is the most common pathological subtype. Accumulating evidence suggests the tumor microenvironment (TME) is correlated with the tumor progress and the patient’s outcome. As the major components of TME, the tumor-infiltrated immune cells and stromal cells have attracted more and more attention. In this study, differentially expressed immune and stromal signature genes were used to construct a TME-related prognostic model for predicting the outcomes of LADC patients.MethodsThe expression profiles of LADC samples with clinical information were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). The differentially expressed genes (DEGs) related to the TME of LADC were identified using TCGA dataset by Wilcoxon rank sum test. The prognostic effects of TME-related DEGs were analyzed using univariate Cox regression. Then, the least absolute shrinkage and selection operator (LASSO) regression was performed to reduce the overfit and the number of genes for further analysis. Next, the prognostic model was constructed by step multivariate Cox regression and risk score of each sample was calculated. Then, survival and Receiver Operating Characteristic (ROC) analyses were conducted to validate the model using TCGA and GEO datasets, respectively. The Kyoto Encyclopedia of Genes and Genomes analysis of gene signature was performed using Gene Set Enrichment Analysis (GSEA). Finally, the overall immune status, tumor purity and the expression profiles of HLA genes of high- and low-risk samples was further analyzed to reveal the potential mechanisms of prognostic effects of the model.ResultsA total of 93 TME-related DEGs were identified, of which 23 DEGs were up-regulated and 70 DEGs were down-regulated. The univariate cox analysis indicated that 23 DEGs has the prognostic effects, the hazard ratio ranged from 0.65 to 1.25 (p < 0.05). Then, seven genes were screened out from the 23 DEGs by LASSO regression method and were further analyzed by step multivariate Cox regression. Finally, a three-gene (ADAM12, Bruton Tyrosine Kinase (BTK), ERG) signature was constructed, and ADAM12, BTK can be used as independent prognostic factors. The three-gene signature well stratified the LADC patients in both training (TCGA) and testing (GEO) datasets as high-risk and low-risk groups, the 3-year area under curve (AUC) of ROC curves of three GEO sets were 0.718 (), 0.646 () and 0.643 (). The GSEA analysis indicated that highly expressed ADAM12, BTK, ERG mainly correlated with the activation of pathways involving in focal adhesion, immune regulation. The immune analysis indicated that the low-risk group has more immune activities and higher expression of HLA genes than that of the high-risk group. In sum, we identified and constructed a three TME-related DEGs signature, which could be used to predict the prognosis of LADC patients.

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Section: Discussionmentioning

confidence: 99%

Identification of prognostic gene signature associated with microenvironment of lung adenocarcinoma

Cheng¹,

Ma²,

Zhou³

2019

PeerJ

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“…GANAB was among proteins most enriched in acidic exosomes implicated in melanoma progression, with high expression levels associated with poor prognosis [54]. It interacts with the short secreted isoform of ADAM12, which is overexpressed in many tumor types [55]. Knockdown of GANAB or PRKCSH, which encode the glucosidase II heterodimer, reduced Wnt3 secretion by 40%-50%, arguing for the role in Wnt signalling [56].…”

Section: Discussionmentioning

confidence: 99%

Cancer-Associated Substitutions in RNA Recognition Motifs of PUF60 and U2AF65 Reveal Residues Required for Correct Folding and 3′ Splice-Site Selection

Královičová

Borovská

Kubíčková

et al. 2020

Cancers

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U2AF65 (U2AF2) and PUF60 (PUF60) are splicing factors important for recruitment of the U2 small nuclear ribonucleoprotein to lariat branch points and selection of 3′ splice sites (3′ss). Both proteins preferentially bind uridine-rich sequences upstream of 3′ss via their RNA recognition motifs (RRMs). Here, we examined 36 RRM substitutions reported in cancer patients to identify variants that alter 3′ss selection, RNA binding and protein properties. Employing PUF60- and U2AF65-dependent 3′ss previously identified by RNA-seq of depleted cells, we found that 43% (10/23) and 15% (2/13) of independent RRM mutations in U2AF65 and PUF60, respectively, conferred splicing defects. At least three RRM mutations increased skipping of internal U2AF2 (~9%, 2/23) or PUF60 (~8%, 1/13) exons, indicating that cancer-associated RRM mutations can have both cis- and trans-acting effects on splicing. We also report residues required for correct folding/stability of each protein and map functional RRM substitutions on to existing high-resolution structures of U2AF65 and PUF60. These results identify new RRM residues critical for 3′ss selection and provide relatively simple tools to detect clonal RRM mutations that enhance the mRNA isoform diversity.

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“…Previous studies on ADAM12S were mainly focused on its proteolytic substrates and interacting proteins. 9,20,44 Several ADAM12S substrates such as E-cadherin, IGFBP-3/5, gelatin, type-IV collagen, and fibronectin have been discovered. 9,15,44 GRP94, α9β1 integrin, and β3 integrin interact with ADAM12 and are implicated in the regulation of cell adhesion, proliferation, and invasion.…”

Section: ■ Discussionmentioning

confidence: 99%

“…ADAM12S was amplified by PCR from the previous constructed pCDF-ADAM12S-FLAG plasmid 20 and inserted into pLVX-IRES-ZsGreen1 vector at the EcoRI/BamHI sites to obtain pLVX-ADAM12S-FLAG-IRES-ZsGreen1 plasmid, which was further validated by Sanger sequencing. pLVX-IRES-ZsGreen1 or pLVX-ADAM12S-FLAG-IRES-ZsGreen1 plasmid was transfected together with the packaging plasmids psPAX2 and pMD2.0G at a ratio of 3:2:1 into HEK293T cells to produce lentiviral particles.…”

Section: Construction Of Plasmids and Stable Cell Linesmentioning

confidence: 99%

“…13,17 ADAM12S also regulates the cell adhesion by binding to the α9β1 integrin or β3 integrin through disintegrin-like domain. 18,19 Our previous studies have screened the interacting proteins for ADAM12S in HeLa cells by quantitative proteomics and identified endoplasmin (GRP94) as its upstream regulator, 20 which is closely associated with the proliferation and metastasis of various cancers. Although many substrates and interacting partners for ADAM12S have been identified, the intracellular proteins regulated by ADAM12S and the involved signaling pathways remain elusive.…”

Section: ■ Introductionmentioning

confidence: 99%

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SILAC Quantitative Proteomics and Biochemical Analyses Reveal a Novel Molecular Mechanism by Which ADAM12S Promotes the Proliferation, Migration, and Invasion of Small Cell Lung Cancer Cells through Upregulating Hexokinase 1

Xiong

Chang

et al. 2019

J. Proteome Res.

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Small cell lung cancer (SCLC) accounts for ∼14% of total lung cancer, which is the worldwide leading cause of morbidity and mortality in cancer. Although SCLC can be treated with chemotherapy and radiotherapy, its 5 year survival rate is still below 7%. Therefore, it is essential to discover new molecules and elucidate the underlying mechanisms modulating the tumorigenesis and metastasis of SCLC for the unmet medical needs. The secreted form of A Disintegrin And Metalloproteinase 12 (ADAM12S) is highly expressed in SCLC and promotes the proliferation, migration, and invasion of SCLC cells. However, the underlying molecular mechanism is still elusive. Using stable isotope labeling by amino acids in cell culture (SILAC)-based quantitative proteomics, we identify 82 ADAM12S-regulated proteins in an SCLC cell line. Our proteomics and biochemical analyses discover that ADAM12S overexpression elevates while ADAM12 knockdown reduces the rate-limiting enzyme hexokinase 1 (HK1) in glycolysis. Through bioinformatics analyses, genetic manipulation, and in vitro assays, we further reveal that ADAM12S promotes the proliferation, colony formation, migration, and invasion of SCLC cells through upregulating HK1. This work links ADAM12S to glucose metabolic pathways in its attribution to the tumorigenesis and metastasis of SCLC cells and might provide valuable information for the exploration of therapeutic intervention for SCLC.

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Quantitative proteomics and biochemical analyses reveal the role of endoplasmin in the regulation of the expression and secretion of A Disintegrin And Metalloproteinase 12

Cited by 4 publications

References 54 publications

Identification of prognostic gene signature associated with microenvironment of lung adenocarcinoma

Identification of prognostic gene signature associated with microenvironment of lung adenocarcinoma

Cancer-Associated Substitutions in RNA Recognition Motifs of PUF60 and U2AF65 Reveal Residues Required for Correct Folding and 3′ Splice-Site Selection

SILAC Quantitative Proteomics and Biochemical Analyses Reveal a Novel Molecular Mechanism by Which ADAM12S Promotes the Proliferation, Migration, and Invasion of Small Cell Lung Cancer Cells through Upregulating Hexokinase 1

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