Cancer Chemotherapy and Pharmacology

, Volume 64, Issue 2, pp 307–316 | Cite as

Stable XIAP knockdown clones of HCT116 colon cancer cells are more sensitive to TRAIL, taxanes and irradiation in vitro

  • Kate Connolly
  • Richard Mitter
  • Morwenna Muir
  • Duncan Jodrell
  • Sylvie Guichard
Original Article

Abstract

Purpose

To develop a model of X-linked inhibitor of apoptosis (XIAP) down regulation in colorectal cancer cell lines. This may be used to determine whether combination strategies have clinical potential.

Methods

A series of clones were developed using short hairpin RNA (shRNA) against XIAP stably expressed in HCT116 cells. XIAP mRNA and protein levels were established by RT-PCR and Immunoblot, respectively. GeneChip microarrays confirmed XIAP knockdown and absence of compensation by other IAP members.

Results

Four XIAP knockdown cell lines show 82–93% reduction in XIAP mRNA and 67–89% reduction in protein when compared to four luciferase control cell lines. XIAP knockdown sensitises cells to rhTRAIL by a factor of 3, to paclitaxel and docetaxel by a factor of >2 and, to a lesser extent, radiotherapy (20% enhancement).

Conclusions

Clinical trials with XIAP antisense continue, and these data suggest combination studies with agents such as rhTRAIL and taxanes should be undertaken.

Keywords

XIAP HCT116 TRAIL Radiotherapy Paclitaxel Docetaxel Microarray 

Abbreviations

AMPK

AMP-activated protein kinase

cIAP1

Cellular inhibitor of apoptosis protein 1

cIAP2

Cellular inhibitor of apoptosis protein 2

DR

Death receptor

IAP

Inhibitor of apoptosis protein

NSCLC

Non-small cell lung cancer

NAIP

Neuronal apoptosis inhibitor protein

MTT

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

RMA

Robust multichip average

RNAi

RNA interference

rhTRAIL

Recombinant human TNF related apoptosis inducing ligand

RT-PCR

Real time polymerase chain reaction

shRNA

Short hairpin RNA

SRB

Sulforhodamine B

XIAP

X-linked inhibitor of apoptosis protein

Notes

Acknowledgments

The authors would like to thank Michael Dodds and Susan Alexander for technical assistance, Aegera Therapeutics Inc for provision of the shRNA constructs and the Cancer Research UK GeneChip microarray service based in the Paterson Institute, University of Manchester for processing the microarray samples.

Conflict of interest statement

K Connolly was funded by Cancer Research UK (Grant number C96/A4743) and her salary was supported in part by Aegera Therapeutics Inc. The Edinburgh Cancer Research Centre received clinical trial support for the Phase I study AEG35156 (XIAP antisense) from Aegera Therapeutics Inc.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Kate Connolly
    • 1
  • Richard Mitter
    • 2
  • Morwenna Muir
    • 1
  • Duncan Jodrell
    • 1
    • 3
  • Sylvie Guichard
    • 1
    • 4
  1. 1.University of Edinburgh Cancer Research CentreEdinburghUK
  2. 2.Bioinformatics and BiostatisticsCancer Research UKLondonUK
  3. 3.Department of Oncology, Addenbrooke’s HospitalUniversity of CambridgeCambridgeUK
  4. 4.AstraZenecaMacclesfieldUK

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