Functional Impact of Collagens on the Activity Directed by the Promoter of the<i>α5</i>Integrin Subunit Gene in Corneal Epithelial Cells

Lake, Jennifer; Zaniolo, Karine; Gingras, Marie-Ève; Couture, Camille; Salesse, Christian; Guérin, Sylvain L.

doi:10.1167/iovs.15-16587

Cited by 7 publications

(13 citation statements)

References 85 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ITGA5 has been widely accepted as involved in the process of invasiveness and tumorigenesis [20,32]. In our research, the expression level of ITGA5 in SiITGA5 group was down-regulated at both mRNA and protein levels, which means that ITGA5 might work as a facilitator in GBM.…”

Section: Discussionmentioning

confidence: 51%

miR-330-5p suppresses glioblastoma cell proliferation and invasiveness through targeting ITGA5

Feng

et al. 2017

Bioscience Reports

View full text Add to dashboard Cite

The present study intended to investigate the biological effects of miR-330-5p on glioblastoma (GBM) cell proliferation and invasiveness by targeting integrin α5 (ITGA5). The expressions of miR-330-5p and ITGA5 mRNA in GBM cell lines (U87, U251, and U373) and normal brain glial cell line (HEB) were detected using RT-qPCR. Protein expression of ITGA5 was examined using Western blot. The present study used MTT assay, colony formation assay, Transwell assay, wound healing assay, and flow cytometry analysis in order to determine the biological functions of GBM cells (including cell proliferation, invasion, migration, apoptosis, and cell cycle). The present study applied dual-luciferase reporter gene assay to identify the target relationship between miR-330-5p and ITGA5. miR-330-5p was low-expressed in GBM cell lines while ITGA5 was high-expressed compared with HEB. miR-330-5p could directly target ITGA5 as well as suppress its expression in GBM cells. Up-regulation of miR-330-5p and down-regulation of ITGA5 both have an inhibitory effect on cell proliferation, invasion, and migration. Meanwhile, they could also promote GBM cell apoptosis. miR-330-5p could suppress proliferation and invasion of GBM cells through targeting ITGA5.

show abstract

Section: Discussionmentioning

confidence: 51%

miR-330-5p suppresses glioblastoma cell proliferation and invasiveness through targeting ITGA5

Feng

et al. 2017

Bioscience Reports

View full text Add to dashboard Cite

show abstract

“…However, none of them have been done in the context of human wound healing of the cornea. The proximal element identified in our study (-82/-203) bears target sites for TBP (TATA-binding protein), AP-1 and both the Sp1 and Sp3 family members (Figure 3A), which we are very familiar with [11,17,18,53,66]. We demonstrated that CLU gene transcription was indeed ensured in part by the binding of the TFs Sp1/Sp3 and AP-1 to overlapping target sites within the basal promoter segment CLU -203/-153.…”

Section: Discussionmentioning

confidence: 85%

“…The significant reduction in the formation of both the AP-1 and Sp1/Sp3 complexes combined to the formation of new, slow-migrating supershifted complexes (SSC) upon addition of a polyclonal antibody that recognizes AP-1 (lane 3), Sp1 (lane 4), or the combination of both Sp1 and Sp3 (lane 5), provided evidence that formation of complexes b and a/c resulted from the recognition of the CLU-203/-/53 labeled probe by AP-1 and Sp1/Sp3, respectively (Figure 4D). Formation of complex d results from the recognition of the labeled probe by non-specific DNA binding proteins, as is often observed in EMSA [17,18,[52][53][54][55][56][57].…”

Section: Members From the Ap1 And Sp1 Families Bind To The Clu Basal Promoter In Hcecsmentioning

confidence: 94%

“…We succeeded in producing two-layer hTECs (epithelium and stroma) made up of primary cultured cells grown on a naturally secreted extracellular matrix that show characteristics very similar to those of the native cornea, including the expression of the epithelial barrier marker ZO-1, the differentiation marker keratins K3/K12, the corneal integrins αvβ6 and α2β1 and integrin subunits β4, α3 and α6 and the subepithelial basement membrane and stromal components laminin V, collagen types I, IV, V and VII, to name a few [9][10][11][12][13]. Over the last 20 years, we used this substitute to study the mechanistic of wound healing [2,9,11,[14][15][16][17][18]. The dynamic of wound closure was found to be very similar between the hTEC and the native cornea [11].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Contribution of the Transcription Factors Sp1/Sp3 and AP-1 to Clusterin Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas

Gross

Le‐Bel

Desjardins

et al. 2021

IJMS

Self Cite

View full text Add to dashboard Cite

In order to reduce the need for donor corneas, understanding of corneal wound healing and development of an entirely tissue-engineered human cornea (hTECs) is of prime importance. In this study, we exploited the hTEC to determine how deep wound healing affects the transcriptional pattern of corneal epithelial cells through microarray analyses. We demonstrated that the gene encoding clusterin (CLU) has its expression dramatically repressed during closure of hTEC wounds. Western blot analyses confirmed a strong reduction in the expression of the clusterin isoforms after corneal damage and suggest that repression of CLU gene expression might be a prerequisite to hTEC wound closure. Transfection with segments from the human CLU gene promoter revealed the presence of three regulatory regions: a basal promoter and two more distal negative regulatory regions. The basal promoter bears DNA binding sites for very potent transcription factors (TFs): Activator Protein-1 (AP-1) and Specificity protein-1 and 3 (Sp1/Sp3). By exploiting electrophoretic mobility shift assays (EMSA), we demonstrated that AP-1 and Sp1/Sp3 have their DNA binding site overlapping with one another in the basal promoter of the CLU gene in hCECs. Interestingly, expression of both these TFs is reduced (at the protein level) during hTEC wound healing, thereby contributing to the extinction of CLU gene expression during that process. The results of this study contribute to a better understanding of the molecular mechanisms accounting for the repression of CLU gene expression during corneal wound healing.

show abstract

“…Some of these key target genes have been shown to play a role in EC in previous studies. As an integrin α subunit, ITGA5 is involved in invasiveness and tumorigenesis (20,21) and is one of the most abundant proteins in the extracellular matrix. Numerous studies have shown that ITGA5 could have a regulatory role in specific cancers by activating focal adhesion kinases to promote cell adhesion and migration (21)(22)(23)(24).…”

Section: Discussionmentioning

confidence: 99%

miR-30b-5p acts as a tumor suppressor microRNA in esophageal squamous cell carcinoma

Xu¹,

Lv²,

Zhang³

et al. 2019

J. Thorac. Dis

View full text Add to dashboard Cite

Background: To study miR-30b-5p expression in esophageal squamous cell carcinoma (ESCC) by comparisons between tumor tissues and matched adjacent non-cancerous tissues to elucidate the correlation between miR-30b-5p expression and ESCC clinical parameters, and to explore the signaling pathways associated with miR-30b-5p and key target genes. Methods: Clinical data, cancer tissues, and adjacent non-cancerous tissues of 32 patients diagnosed with ESCC were collected from Taizhou Hospital of Zhejiang Province. The expression levels of miR-30b-5p were determined by real-time polymerase chain reaction (RT-PCR). mRNA data for ESCC tissues and normal tissues, and clinical materials of patients with ESCC were obtained from the Gene Expression Omnibus (GEO) database and The Cancer Genome Atlas (TCGA). Associations between miR-30b-5p expression and clinical features of patients with ESCC and overall survival were explored. A bioinformatics analysis was performed to determine the pathways and key miR-30b-5p targets associated with ESCC.Additionally, a cytological experiment was performed to evaluate the biological functions of miR-30b-5p.Finally, correlations between miR-30b-5p and key targets involved in PI3K/Akt signaling pathways were validated by western blotting.Results: The expression level of miR-30b-5p in the 32 ESCC tissues was significantly lower than that in adjacent normal tissues (P<0.01) and was significantly disparate in the T stage, with higher expression in T1 than in T2 (P<0.05). Among the patients with higher expression levels of miR-30b-5p in ESCC tissues than in adjacent normal tissues, patients with higher expression of miR-30b-5p had a better prognosis (P<0.05).An analysis of gene chip data from the GEO database showed similar results. A gene enrichment analysis indicated a series of pathways that may be associated with the downregulation of miR-30b-5p, including focal adhesion, ECM-receptor interaction, and PI3K/Akt signaling pathways. Seven key target genes (PDGFRB, VIM, ITGA5, ACTN1, THBS2, SERPINE1, and RUNX2) were identified; these were found to be upregulated in ESCC tissues and were negatively correlated with miR-30b-5p. Functional experiments showed that miR-30b-5p attenuated migration (P<0.01) and invasion (P<0.05) in the Eca109 cell line. Moreover, the levels of ITGA5, PDGFRB, p-PI3K, and p-AKT, which are involved in the PI3K/Akt signaling pathway, were decreased in the miR-30b-5p-overexpressing Eca109 cell line. Conclusions: Upregulated miR-30b-5p may inhibit migration and invasion in ESCC by targeting ITGA5, PDGFRB, and signaling pathways, such as PI3K/Akt, involved in ESCC regulation. Our results indicate that miR-30b-5p plays an important role in the occurrence and progression of ESCC and is a potential therapeutic target.

show abstract

Functional Impact of Collagens on the Activity Directed by the Promoter of theα5Integrin Subunit Gene in Corneal Epithelial Cells

Abstract: Collagens considerably suppressed α5 gene expression in CECs, suggesting that during wound healing, they may interfere with the influence FN exerts on CECs by altering their adhesive and migratory properties through a mechanism involving a reduction in α5 gene expression.

Cited by 7 publications

References 85 publications

miR-330-5p suppresses glioblastoma cell proliferation and invasiveness through targeting ITGA5

miR-330-5p suppresses glioblastoma cell proliferation and invasiveness through targeting ITGA5

Contribution of the Transcription Factors Sp1/Sp3 and AP-1 to Clusterin Gene Expression during Corneal Wound Healing of Tissue-Engineered Human Corneas

miR-30b-5p acts as a tumor suppressor microRNA in esophageal squamous cell carcinoma

Contact Info

Product

Resources

About