The role of the cholecystokinin 2 receptor in cancer
The gastrointestinal (GI) hormone, gastrin, promotes cancer progression and its down-regulation has been linked to reduced cancer stem cell numbers. Gastrin acts through the cholecystokinin-2 receptor (CCK-2R) and its biological effects are blocked by CCK-2R inhibitors. We investigated the regulatio...
| Main Author: | |
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| Format: | Thesis (University of Nottingham only) |
| Language: | English English |
| Published: |
2011
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| Online Access: | https://eprints.nottingham.ac.uk/12705/ |
| _version_ | 1848791563648368640 |
|---|---|
| author | Mayne, Cerys Mary |
| author_facet | Mayne, Cerys Mary |
| author_sort | Mayne, Cerys Mary |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | The gastrointestinal (GI) hormone, gastrin, promotes cancer progression and its down-regulation has been linked to reduced cancer stem cell numbers. Gastrin acts through the cholecystokinin-2 receptor (CCK-2R) and its biological effects are blocked by CCK-2R inhibitors. We investigated the regulation of the CCK-2R and its potential role in promoting survival of cancer stem cells (CSC).
A panel of cancer cell-lines, including GI, glioblastoma and lung, with CCK-2R-transfected cells as a positive control, were grown either as monolayers, or, to provide a 3D in vitro tumour model, as spheres. Linear-after-the-Exponential (LATE)-PCR was used to quantify CCK-2R gene expression and this was validated using siRNAs. Flow cytometry was used to investigate receptor protein expression. Activity of CCK-2R promoter reporters was quantified using luciferase assays.
LATE-PCR for CCK-2R gene expression is 10,000-fold more sensitive than the Taqman-based assay, and provides a highly precise method for detection of genes which have important biological functions but low expression. This assay showed that primary non-small-cell lung tumours have significantly more expression than normal lung tissue, indicating a potential therapeutic marker. CCK-2R siRNAs resulted in up to 97% (p<0.05) knockdown of the receptor in cancer cells, confirming the specificity of LATE-PCR and offering a therapeutic possibility.
The CCK-2R promoter constructs were active in lung, glioma and colorectal cancer cell-lines, demonstrating a potential drug target; however, transcriptional activity did not correlate with gene expression suggesting post-transcriptional or translational regulation is a factor affecting CCK-2R expression.
Flow cytometry suggests the presence of a small population of cells within each of these cell-lines which expresses CCK-2R very highly, which was not correlated to CSC markers. However, CCK-2R expression was enriched when cells were grown as spheres, and inhibition caused a delay in sphere-forming, implying that the CCK-2R may play a role in tumour, and CSC, expansion. Thus, CCK2R provides a potential target for therapeutic intervention in cancer. |
| first_indexed | 2025-11-14T18:30:30Z |
| format | Thesis (University of Nottingham only) |
| id | nottingham-12705 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| language | English English |
| last_indexed | 2025-11-14T18:30:30Z |
| publishDate | 2011 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-127052025-02-28T11:20:54Z https://eprints.nottingham.ac.uk/12705/ The role of the cholecystokinin 2 receptor in cancer Mayne, Cerys Mary The gastrointestinal (GI) hormone, gastrin, promotes cancer progression and its down-regulation has been linked to reduced cancer stem cell numbers. Gastrin acts through the cholecystokinin-2 receptor (CCK-2R) and its biological effects are blocked by CCK-2R inhibitors. We investigated the regulation of the CCK-2R and its potential role in promoting survival of cancer stem cells (CSC). A panel of cancer cell-lines, including GI, glioblastoma and lung, with CCK-2R-transfected cells as a positive control, were grown either as monolayers, or, to provide a 3D in vitro tumour model, as spheres. Linear-after-the-Exponential (LATE)-PCR was used to quantify CCK-2R gene expression and this was validated using siRNAs. Flow cytometry was used to investigate receptor protein expression. Activity of CCK-2R promoter reporters was quantified using luciferase assays. LATE-PCR for CCK-2R gene expression is 10,000-fold more sensitive than the Taqman-based assay, and provides a highly precise method for detection of genes which have important biological functions but low expression. This assay showed that primary non-small-cell lung tumours have significantly more expression than normal lung tissue, indicating a potential therapeutic marker. CCK-2R siRNAs resulted in up to 97% (p<0.05) knockdown of the receptor in cancer cells, confirming the specificity of LATE-PCR and offering a therapeutic possibility. The CCK-2R promoter constructs were active in lung, glioma and colorectal cancer cell-lines, demonstrating a potential drug target; however, transcriptional activity did not correlate with gene expression suggesting post-transcriptional or translational regulation is a factor affecting CCK-2R expression. Flow cytometry suggests the presence of a small population of cells within each of these cell-lines which expresses CCK-2R very highly, which was not correlated to CSC markers. However, CCK-2R expression was enriched when cells were grown as spheres, and inhibition caused a delay in sphere-forming, implying that the CCK-2R may play a role in tumour, and CSC, expansion. Thus, CCK2R provides a potential target for therapeutic intervention in cancer. 2011-12-13 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/12705/1/Cerys_Thesis_locked.pdf application/pdf en arr https://eprints.nottingham.ac.uk/12705/2/Cerys_Mayne_Thesis_Appendix.pdf Mayne, Cerys Mary (2011) The role of the cholecystokinin 2 receptor in cancer. PhD thesis, University of Nottingham. |
| spellingShingle | Mayne, Cerys Mary The role of the cholecystokinin 2 receptor in cancer |
| title | The role of the cholecystokinin 2 receptor in cancer |
| title_full | The role of the cholecystokinin 2 receptor in cancer |
| title_fullStr | The role of the cholecystokinin 2 receptor in cancer |
| title_full_unstemmed | The role of the cholecystokinin 2 receptor in cancer |
| title_short | The role of the cholecystokinin 2 receptor in cancer |
| title_sort | role of the cholecystokinin 2 receptor in cancer |
| url | https://eprints.nottingham.ac.uk/12705/ |