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dc.contributor.authorJoshi, Rajeshree
dc.contributor.authorTawfik, Amany
dc.contributor.authorEdeh, Nneka
dc.contributor.authorMcCloud, Veronica
dc.contributor.authorLooney, Stephen W.
dc.contributor.authorLewis, Jill
dc.contributor.authorHsu, Stephen
dc.contributor.authorOgbureke, Kalu U.E.
dc.date.accessioned2012-10-26T16:26:52Z
dc.date.available2012-10-26T16:26:52Z
dc.date.issued2010-11-12en_US
dc.identifier.citationPLoS One. 2010 Nov 12; 5(11):e13974en_US
dc.identifier.issn1932-6203en_US
dc.identifier.pmid21103065en_US
dc.identifier.doi10.1371/journal.pone.0013974en_US
dc.identifier.urihttp://hdl.handle.net/10675.2/609
dc.description.abstractBackground: We determined recently that dentin sialophosphoprotein (DSPP), a member of the SIBLING (Small integrin-binding ligand N-linked glycoproteins) family of phosphoglycoproteins, is highly upregulated in human oral squamous cell carcinomas (OSCCs) where upregulation is associated with tumor aggressiveness. To investigate the effects of DSPP-silencing on the tumorigenic profiles of the oral cancer cell line, OSC2, short-hairpin RNA (shRNA) interference was employed to silence DSPP in OSC2 cells.
dc.description.abstractMethodology/Principal Findings: Multiple regions of DSPP transcript were targeted for shRNA interference using hDSPshRNA lentiviral particles designed to silence DSPP gene expression. Control shRNA plasmid encoding a scrambled sequence incapable of degrading any known cellular mRNA was used for negative control. Following puromycin selection of stable lines of DSSP-silenced OSC2 cells, phenotypic hallmarks of oral carcinogenesis were assayed by western blot and RT-PCR analyses, MTT (cell-viability), colony-formation, modified Boyden-Chamber (migration and invasion), and flow cytometry (cell-cycle and apoptosis) analyses. DSPP-silenced OSC2 cells showed altered cell morphology, reduced viability, decreased colony-formation ability, decreased migration and invasion, G0/G1 cell-cycle arrest, and increased tumor cell sensitivity to cisplatin-induced apoptosis. Furthermore, MMP-2, MMP-3, MMP-9, VEGF, Ki-67, p53, and EGFR were downregulated. There was a direct correlation between the degree of DSPP-silencing and MMP suppression, as indicated by least squares regression: MMP-2 {(y = 0.850x, p,0.001) (y = 1.156x, p,0.001)}, MMP-3 {(y = 0.994x, p,0.001) (y = 1.324x, p = 0.004)}, and MMP-9 {(y = 1.248x, p = 0.005, y = 0.809, p = 0.013)}.
dc.description.abstractConclusions/Significance: DSPP-silencing in OSC2 cell decreased salient hallmarks of oral tumorigenesis and provides the first functional evidence of a potential key role for DSPP in oral cancer biology. The down-regulation of MMP-2, MMP-3, MMP-9, p53 and VEGF in DSPP-silenced OSC2 cells provides a significant functional/molecular framework for deciphering the mechanisms of DSPP activities in oral cancer biology.
dc.rightsJoshi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.en_US
dc.subjectResearch Articleen_US
dc.subjectMolecular Biologyen_US
dc.subjectCell Biology/Cell Growth and Divisionen_US
dc.subjectMolecular Biology/DNA Methylationen_US
dc.subjectMolecular Biology/DNA Repairen_US
dc.subjectOncology/Head and Neck Cancersen_US
dc.subjectPathology/Histopathologyen_US
dc.subjectPathology/Molecular Pathologyen_US
dc.subjectPathology/Surgical Pathologyen_US
dc.subjectCell Biology/Extra-Cellular Matrixen_US
dc.subject.meshAnimalsen_US
dc.subject.meshAntineoplastic Agentsen_US
dc.subject.meshApoptosisen_US
dc.subject.meshBlotting, Westernen_US
dc.subject.meshCell Lineen_US
dc.subject.meshCell Line, Tumoren_US
dc.subject.meshCell Movementen_US
dc.subject.meshCisplatinen_US
dc.subject.meshExtracellular Matrix Proteinsen_US
dc.subject.meshFemaleen_US
dc.subject.meshHumansen_US
dc.subject.meshKeratinocytesen_US
dc.subject.meshMatrix Metalloproteinase 2en_US
dc.subject.meshMatrix Metalloproteinase 3en_US
dc.subject.meshMatrix Metalloproteinase 9en_US
dc.subject.meshMiceen_US
dc.subject.meshMice, Inbred BALB Cen_US
dc.subject.meshMice, Nudeen_US
dc.subject.meshMouth Neoplasmsen_US
dc.subject.meshNeoplasms, Experimentalen_US
dc.subject.meshPhosphoproteinsen_US
dc.subject.meshRNA Interferenceen_US
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen_US
dc.subject.meshSialoglycoproteinsen_US
dc.subject.meshTransplantation, Heterologousen_US
dc.subject.meshTumor Suppressor Protein p53en_US
dc.subject.meshUp-Regulationen_US
dc.titleDentin Sialophosphoprotein (DSPP) Gene-Silencing Inhibits Key Tumorigenic Activities in Human Oral Cancer Cell Line, OSC2en_US
dc.typeArticleen_US
dc.identifier.pmcidPMC2980487en_US
dc.contributor.corporatenameDepartment of Oral Biology
dc.contributor.corporatenameDepartment of Pathology
dc.contributor.corporatenameDepartment of Oral Health and Diagnostic Sciences
dc.contributor.corporatenameCollege of Graduate Studies
dc.contributor.corporatenameDepartment of Biostatistics and Epidemiology
refterms.dateFOA2019-04-09T22:51:05Z
html.description.abstractBackground: We determined recently that dentin sialophosphoprotein (DSPP), a member of the SIBLING (Small integrin-binding ligand N-linked glycoproteins) family of phosphoglycoproteins, is highly upregulated in human oral squamous cell carcinomas (OSCCs) where upregulation is associated with tumor aggressiveness. To investigate the effects of DSPP-silencing on the tumorigenic profiles of the oral cancer cell line, OSC2, short-hairpin RNA (shRNA) interference was employed to silence DSPP in OSC2 cells.
html.description.abstractMethodology/Principal Findings: Multiple regions of DSPP transcript were targeted for shRNA interference using hDSPshRNA lentiviral particles designed to silence DSPP gene expression. Control shRNA plasmid encoding a scrambled sequence incapable of degrading any known cellular mRNA was used for negative control. Following puromycin selection of stable lines of DSSP-silenced OSC2 cells, phenotypic hallmarks of oral carcinogenesis were assayed by western blot and RT-PCR analyses, MTT (cell-viability), colony-formation, modified Boyden-Chamber (migration and invasion), and flow cytometry (cell-cycle and apoptosis) analyses. DSPP-silenced OSC2 cells showed altered cell morphology, reduced viability, decreased colony-formation ability, decreased migration and invasion, G0/G1 cell-cycle arrest, and increased tumor cell sensitivity to cisplatin-induced apoptosis. Furthermore, MMP-2, MMP-3, MMP-9, VEGF, Ki-67, p53, and EGFR were downregulated. There was a direct correlation between the degree of DSPP-silencing and MMP suppression, as indicated by least squares regression: MMP-2 {(y = 0.850x, p,0.001) (y = 1.156x, p,0.001)}, MMP-3 {(y = 0.994x, p,0.001) (y = 1.324x, p = 0.004)}, and MMP-9 {(y = 1.248x, p = 0.005, y = 0.809, p = 0.013)}.
html.description.abstractConclusions/Significance: DSPP-silencing in OSC2 cell decreased salient hallmarks of oral tumorigenesis and provides the first functional evidence of a potential key role for DSPP in oral cancer biology. The down-regulation of MMP-2, MMP-3, MMP-9, p53 and VEGF in DSPP-silenced OSC2 cells provides a significant functional/molecular framework for deciphering the mechanisms of DSPP activities in oral cancer biology.


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