The <scp>CRISPR</scp>/Cas system inhibited the pro‐oncogenic effects of alternatively spliced fibronectin extra domain A via editing the genome in salivary adenoid cystic carcinoma cells

Hc, Wang; Yang, Yanwen; Sy, Xu; Peng, Jing; Jh, Jiang; Cy, Li

doi:10.1111/odi.12323

Cited by 17 publications

(50 citation statements)

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“…The fibroblasts were successfully isolated from the surgical specimens of 20 in the 22 cases, and the cells from each cases were routinely cultured for in vitro analysis, using α‐modified Eagle's medium (α‐MEM; GIBCO, Grand Island, NY, USA; Wang & Li , Wang et al 2015a, Wang et al 2015b). Raw264.7, the osteoclast precursor, was grown in Dulbecco's modified Eagle's medium (DMEM) (GIBCO).…”

Section: Methodsmentioning

confidence: 99%

“…According to previous studies, fibroblasts isolated from the fibrous capsule of odontogenic cysts induce more osteoclasts than do normal controls (Wang & Li , Wang et al 2015a, Wang et al 2015b), possibly attributed to the differentiation of myofibroblasts, which are activated during chronic inflammation (Kouhsoltani et al ) or inappropriate wound healing (Lemanska‐Perek et al ). Chronic inflammation may initiate radicular cysts that can also activate fibroblasts in capsules, generating extracellular matrix (ECM; Jiang et al ), cellular factors and metabolic products (de Moraes et al , Tekkesin et al , Bernardi et al ), leading to osteoclastogenesis and bone destruction, as illustrated by radiolucent periapical areas.…”

Section: Introductionmentioning

confidence: 98%

“…Radicular cysts are initiated by chronic inflammation, and inflammatory factors within the fibrous capsules tend to stimulate generation of EDB + FN and CS1‐FN (Boyle et al , Khan et al , Liu et al ). Considering the participation of FN isoforms in tumour aggressiveness (Kamarajan et al , Wang et al 2015a, Wang et al 2015b), bone destruction resulting from odontogenic cysts may be affected by FN isoforms as well.…”

Section: Introductionmentioning

confidence: 99%

“…Since the three main splice variants, EDA + FN, EDB + FN and CS1‐FN, are ligands of integrin receptors and Toll‐like receptor 4 (TLR4), interactions between FN isoforms and receptors in pre‐osteoclasts may directly affect osteoclastogenesis (Doddapattar et al , Jiang et al ), whilst their interaction in other cells could alter the expression of osteoclastogenic genes, such as vascular endothelial growth factor (VEGF) or IL‐17 (Liu et al , Lv et al ), by which osteoclastogenesis can be indirectly influenced. Therefore, this study explored the effects of FN isoforms on bone destruction in radicular cysts, as well as the underlying mechanism through antibody blocking and gene editing using the type II bacterial clustered, regularly interspaced, palindromic repeats (CRISPR)‐associated (Cas) system (Kohan et al , Wang et al 2015a, Wang et al 2015b).…”

Section: Introductionmentioning

confidence: 99%

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The extra domain A of fibronectin facilitates osteoclastogenesis in radicular cysts through vascular endothelial growth factor

et al. 2019

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Aim To analyse the effects of the alternatively spliced fibronectin (FN) gene and its isoforms on osteoclastogenesis in radicular cysts. Methodology Specimens of radicular cysts were collected surgically from 22 patients whose radiolucent periapical areas were measured on digital panoramic radiographs before surgery. The associations between the radiolucent areas and FN isoforms, vascular endothelial growth factor (VEGF) expression or micro‐vessel density, as well as the relationships amongst them, were analysed by immunohistochemical staining using the antibodies IST‐9, BC‐1, P1F11, VEGF and CD34. Fibroblasts isolated from those specimens were used to induce Trap + MNCs, and the effects of induction were assessed by blocking FN containing extra domain A (EDA + FN), COX‐2 or VEGF in vitro. The effects of EDA exon knockout using CRISPR/Cas system were also assessed. Quantitative PCR was used to analyse relative expression of FN isoforms and osteoclastogenic genes. Data were analysed using linear regression, Spearman's rank correlation analysis, chi‐square test and Student's t‐test; P < 0.05 was considered significant. Results Micro‐vessel density and EDA + FN staining were positively associated with the size of radiolucent periapical areas (mm2; P < 0.05), consistent with a positive association between Trap + MNCs and VEGF expression in fibroblasts (P < 0.05). Blocking the interaction between EDA + FN and fibroblasts inhibited Trap + MNC formation. In addition, EDA exon knockout decreased VEGF expression and inhibited Trap + MNC formation to the extent of blocking VEGF by bevacizumab, but osteoclastogenic induction was restored by recombinant VEGF. Using retrospective clinicopathological data, VEGF staining was shown to be positively associated with EDA + FN staining, micro‐vessel density and the size of radiolucent areas (P < 0.05). Conclusion In fibrous capsules of radicular cysts, the alternatively spliced isoform EDA + FN generated by fibroblasts stimulated VEGF expression via an autocrine effect and then facilitated osteoclastogenesis. Both blockage of VEGF and EDA exon knockout could be used to inhibit bone destruction.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

The extra domain A of fibronectin facilitates osteoclastogenesis in radicular cysts through vascular endothelial growth factor

et al. 2019

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“…Total protein was extracted from cells grown in complete medium, and supernatants were collected from cells and tissues [15]. Proteins were separated by 12% SDS-PAGE and transferred to nitrocellulose filter membranes, which were probed overnight at 4 °C with the following antibodies: anti-MDM2 (1:1000, ab38618), anti-p53 (phosopho ser15) (1:500, ab1431), and antiBax (1:1000, ab182734) (Abcam, USA).…”

Section: Western Blotmentioning

confidence: 99%

MALAT1 Activates the P53 Signaling Pathway by Regulating MDM2 to Promote Ischemic Stroke

Zhang

Wang

et al. 2018

Cell Physiol Biochem

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Background/Aims: This study focused on evaluating the effect of MALAT1 and MDM2 on ischemic stroke through regulation of the p53 signaling pathway. Materials: Bioinformatics analysis was performed to identify abnormally expressed lncRNAs, mRNAs and their associated pathways. Oxygen-glucose deprivation/reoxygenation (OGD/R) in cells and middle cerebral artery occlusion/reperfusion (MCAO/R) in mice were performed to simulate an ischemic stroke environment. Western blot and qRT-PCR were used to examine lncRNA expression and mRNA levels. Fluorescence in situ hybridization (FISH) LncRNA was used to locate mRNA. MTT and flow cytometry were performed to examine cell proliferation and apoptosis. Finally, immunohistochemistry was used to observe the expression of genes in vivo. Results: MALAT1 and MDM2, which exhibit strong expression in stroke tissues, were subjected to bioinformatics analysis, and the p53 pathway was chosen for further study. MALAT1, MDM2 and p53 signaling pathway-related proteins were all up regulated in OGD/R cells. Furthermore, Malat1, Mdm2 and p53 pathway related-proteins were also up regulated in MCAO/R mice. Both MALAT1 and MDM2 were localized in the nuclei. Down regulation of MALAT1 and MDM2 enhanced cell proliferation ability and reduced apoptosis, resulting in decreased infarct size in MCAO/R brains. Conclusion: These results indicate that MALAT1/MDM2/p53 signaling pathway axis may provide more effective clinical therapeutic strategy for patients with ischemic stroke.

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Prospective Advances in Genome Editing Investigation

Isola

2022

Advances in Experimental Medicine and Biology

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The CRISPR/Cas system inhibited the pro‐oncogenic effects of alternatively spliced fibronectin extra domain A via editing the genome in salivary adenoid cystic carcinoma cells

Cited by 17 publications

References 44 publications

The extra domain A of fibronectin facilitates osteoclastogenesis in radicular cysts through vascular endothelial growth factor

The extra domain A of fibronectin facilitates osteoclastogenesis in radicular cysts through vascular endothelial growth factor

MALAT1 Activates the P53 Signaling Pathway by Regulating MDM2 to Promote Ischemic Stroke

Prospective Advances in Genome Editing Investigation

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