dc.contributor.author |
Anchan, Akshata |
|
dc.contributor.author |
Martin, Olivia |
|
dc.contributor.author |
Hucklesby, James JW |
|
dc.contributor.author |
Finlay, Graeme |
|
dc.contributor.author |
Johnson, Rebecca H |
|
dc.contributor.author |
Robilliard, Laverne D |
|
dc.contributor.author |
O'Carroll, Simon J |
|
dc.contributor.author |
Angel, Catherine E |
|
dc.contributor.author |
Graham, E Scott |
|
dc.coverage.spatial |
Switzerland |
|
dc.date.accessioned |
2021-03-16T01:46:28Z |
|
dc.date.available |
2021-03-16T01:46:28Z |
|
dc.date.issued |
2020-11 |
|
dc.identifier.citation |
International journal of molecular sciences 21(21) Nov 2020 |
|
dc.identifier.issn |
1422-0067 |
|
dc.identifier.uri |
https://hdl.handle.net/2292/54710 |
|
dc.description.abstract |
We have recently demonstrated that invasive melanoma cells are capable of disrupting the brain endothelial barrier integrity. This was shown using ECIS biosensor technology, which revealed rapid disruption via the paracellular junctions. In this paper, we demonstrate that melanoma cells secrete factors (e.g., cytokines) that weaken the endothelial barrier integrity. Through proteome profiling, we attempt to identify the barrier-disrupting cytokines. Melanoma conditioned media were collected from three New Zealand melanoma lines. ECIS technology was used to assess if the conditioned media disrupted the endothelial barrier independent of the melanoma cells. The melanoma cell secretome was assessed using cytometric bead array (CBA), Luminex immunoassay and multiplex Proteome Profilers, to detect the expression of secretory proteins, which may facilitate metastasis. Finally, ECIS technology was used to assess the direct effects of secreted proteins identified as candidates from the proteome screens. We show that melanoma-conditioned media significantly disrupted the brain endothelial barrier, however, to a much lesser extent than the cells from which they were collected. Cytokine and proteome profiling of the conditioned media showed evidence of high concentrations of approximately 15 secreted proteins (including osteopontin, IL-8, GDF-15, MIF and VEGF). These 15 secreted proteins were expressed variably across the melanoma lines. Surprisingly, the addition of these individually to the brain endothelial cells did not substantially affect the barrier integrity. ANGPTL-4 and TGFβ were also produced by the melanoma cells. Whilst TGFβ-1 had a pronounced effect on the barrier integrity, surprisingly ANGPTL-4 did not. However, its C-terminal fragment did and within a very similar period to the conditioned media, albeit not to the same extent. Herein we show that melanoma cells produce a wide-range of soluble factors at high concentrations, which most likely favour support or survival of the cancer cells. Most of these, except for TGFβ-1 and the C-terminal fragment of ANGPTL-4, did not have an impact on the integrity of the brain endothelial cells. |
|
dc.format.medium |
Electronic |
|
dc.language |
eng |
|
dc.publisher |
MDPI AG |
|
dc.relation.ispartofseries |
International journal of molecular sciences |
|
dc.rights |
Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. |
|
dc.rights.uri |
https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm |
|
dc.rights.uri |
https://creativecommons.org/licenses/by/4.0/ |
|
dc.subject |
ANGPTL-4 |
|
dc.subject |
ECIS technology |
|
dc.subject |
TGFβ |
|
dc.subject |
brain endothelial cells |
|
dc.subject |
cytokine profiling |
|
dc.subject |
melanoma |
|
dc.subject |
metastasis |
|
dc.subject |
Science & Technology |
|
dc.subject |
Life Sciences & Biomedicine |
|
dc.subject |
Physical Sciences |
|
dc.subject |
Biochemistry & Molecular Biology |
|
dc.subject |
Chemistry, Multidisciplinary |
|
dc.subject |
Chemistry |
|
dc.subject |
melanoma |
|
dc.subject |
brain endothelial cells |
|
dc.subject |
ECIS technology |
|
dc.subject |
cytokine profiling |
|
dc.subject |
metastasis |
|
dc.subject |
TGFβ |
|
dc.subject |
ANGPTL-4 |
|
dc.subject |
EPITHELIAL-MESENCHYMAL TRANSITION |
|
dc.subject |
GROWTH-FACTOR |
|
dc.subject |
TGF-BETA |
|
dc.subject |
VASCULAR-PERMEABILITY |
|
dc.subject |
TUMOR-GROWTH |
|
dc.subject |
EXPRESSION |
|
dc.subject |
RECRUITMENT |
|
dc.subject |
METASTASIS |
|
dc.subject |
CANCER |
|
dc.subject |
SPARC |
|
dc.subject |
0399 Other Chemical Sciences |
|
dc.subject |
0604 Genetics |
|
dc.subject |
0699 Other Biological Sciences |
|
dc.title |
Analysis of Melanoma Secretome for Factors That Directly Disrupt the Barrier Integrity of Brain Endothelial Cells. |
|
dc.type |
Journal Article |
|
dc.identifier.doi |
10.3390/ijms21218193 |
|
pubs.issue |
21 |
|
pubs.begin-page |
8193 |
|
pubs.volume |
21 |
|
dc.date.updated |
2021-02-11T19:22:36Z |
|
dc.rights.holder |
Copyright: The authors |
en |
pubs.author-url |
https://www.ncbi.nlm.nih.gov/pubmed/33139674 |
|
pubs.publication-status |
Published |
|
dc.rights.accessrights |
http://purl.org/eprint/accessRights/OpenAccess |
en |
pubs.subtype |
research-article |
|
pubs.subtype |
Journal Article |
|
pubs.elements-id |
824054 |
|
dc.identifier.eissn |
1422-0067 |
|
dc.identifier.pii |
ijms21218193 |
|
pubs.number |
ARTN 8193 |
|
pubs.online-publication-date |
2020-11-1 |
|