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| Status |
Public on Dec 01, 2008 |
| Title |
Stable XIAP knockdown in HCT116 colon cancer cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
X-linked inhibitor of apoptosis (XIAP) is the most potent endogenous caspase inhibitor preventing cell death via caspase-9, -7 and -3 (initiator and executioner caspase pathways). Using short hairpin RNA (shRNA) against XIAP, stably expressed in a parent HCT116 human colon cancer cell line, a series of clones have been developed. XIAP mRNA levels were established by RT-PCR, the four X (XIAP knockdown) clonal cell lines show 82-93% reduction in XIAP mRNA when compared to the four L (luciferase control) cell lines. Immunoblot analysis showed a 67-89% reduction in XIAP protein in X cell lines compared to L. RNA was analysed by microarray and XIAP knockdown was confirmed in 7 probe sets, there was no significant compensation of other IAP family members. XIAP knockdown induced a 2-fold increase in the basal level of apoptosis without modification of caspase 3/7 activity. Finally, XIAP knockdown sensitises cells to radiotherapy by 20%, to recombinant TRAIL by a 3-fold factor, and to paclitaxel and docetaxel by >2 fold factor. Future work should focus on targeted agents such as rhTRAIL in combination with strategies to down regulate XIAP. XIAP antisense is now in clinical development in oncology.
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| Overall design |
Human HCT116 colon cancer cell lines were transfected with either XIAP (X) or luciferase (L) shRNA. The expression profiles of both were compared at early (p4) and late (p8) passage.
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| Contributor(s) |
Connolly KC, Mitter R, Muir M, Jodwell DI, Guichard SM |
| Citation(s) |
19034449 |
| Submission date |
May 30, 2008 |
| Last update date |
Mar 25, 2019 |
| Contact name |
Kathryn Clare Connolly |
| Organization name |
University of Edinburgh
|
| Department |
Cancer Research Centre
|
| Lab |
Pharmacology and Drug Development Group
|
| Street address |
Crewe Road South
|
| City |
Edinburgh |
| ZIP/Postal code |
EH4 2XR |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL570 |
[HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array |
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| Samples (18)
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| Relations |
| BioProject |
PRJNA106125 |
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