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dc.contributor.advisorMcLellan, Alexander Donald
dc.contributor.authorCoutinho, Frazer Paul
dc.date.available2014-02-23T19:45:34Z
dc.date.copyright2014
dc.identifier.citationCoutinho, F. P. (2014). An investigation of CD147 expression in apoptotic vesicles (Thesis, Master of Science). University of Otago. Retrieved from http://hdl.handle.net/10523/4607en
dc.identifier.urihttp://hdl.handle.net/10523/4607
dc.description.abstractOur laboratory has recently found that glycoprotein CD147 (Basigin, EMMPRIN) is highly enriched on the surface of apoptotic vesicles (Apo-V) isolated from EL4 and B16 tumour cells. The CD147 protein can appear at a range of sizes due to the presence of multiple isoforms and glycosylation processes. CD147 has been described as an anti-apoptotic molecule and is involved in the invasion and metastasis of metastatic cancers. Apoptotic vesicles (Apo-V) have been shown to bind the CD169 sialic acid-receptor expressed by macrophages in the sub-capsular sinus of lymph nodes and the marginal zone of the spleen. It has been suggested that this interaction of vesicles with the CD169 receptor may have the potential to suppress immune responses. In this investigation, the expression and role of CD147 in Apo-V was observed. After many failed attempts to knockdown CD147 expression in EL4 cells using small hairpin RNA (shRNA) techniques, an alternate pathway was undertaken using B16 melanoma cells. The observed knockdown in cellular fractions of B16 was also replicated in corresponding Apo-V fractions. No significant changes in Apo-V production in the CD147 shRNA knock-down B16 cell line were observed, neither were there major changes in Coomassie blue bands as analysed by polyacrylamide gel electrophoresis. This suggested that CD147 knockdown did not affect apoptotic vesicle production quality. The binding of Apo-V to spleen and lymph node sections was not affected by the knockdown of CD147, suggesting that the CD147 glycoprotein is not the major ligand for CD169 in lymphatic tissues. To further investigate the potential of CD147 to bind CD169, a CD169-Fc-FLAG protein was produced. However, no specific binding of CD169-Fc-FLAG to CD147 was detected. Since CD147 has been implicated in tumour metastasis, the metastatic potential of CD147 knockdown B16 cell lines was next tested to confirm the knockdown. This resulted in functional changes in the cell line behaviour. However, no conclusive evidence for an effect of CD147 knockdown on metastasis was noted.
dc.format.mimetypeapplication/pdf
dc.language.isoen
dc.publisherUniversity of Otago
dc.rightsAll items in OUR Archive are provided for private study and research purposes and are protected by copyright with all rights reserved unless otherwise indicated.
dc.subjectApo-V
dc.subjectapoptotic
dc.subjectvesicles
dc.subjectCD147
dc.subjectCD169
dc.subjectcancer
dc.subjectmelanoma
dc.subjectB16
dc.subjectEL4
dc.subjecttumour
dc.subjectmetastasis
dc.subjecttransfection
dc.subjectknockdown
dc.subjectGFP
dc.subjectFLAG
dc.subjectspleen
dc.subjectlymph
dc.subjectnode
dc.subjectimmunology
dc.subjectimmunity
dc.subjectglycoproteins
dc.titleAn investigation of CD147 expression in apoptotic vesicles
dc.typeThesis
dc.date.updated2014-02-21T22:07:21Z
dc.language.rfc3066en
thesis.degree.disciplineMicrobiology and Immunology
thesis.degree.nameMaster of Science
thesis.degree.grantorUniversity of Otago
thesis.degree.levelMasters
otago.openaccessOpen
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