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Short hairpin RNAs targeting Bcl-xL modulate senescence and apoptosis following SN-38 and irinotecan exposure in a colon cancer model

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Abstract

Bcl-xL is an anti-apoptotic protein over-expressed in colorectal cancers acting on both the intrinsic and extrinsic pathways. We stably expressed four different short hairpin RNA (pSNG-xL1-4) targeting Bcl-xL in HCT 116 cells. HCT 116 pSNG-xL#1 produced a modest (30%) decrease in Bcl-xL expression whilst Bcl-2 levels were similar to the parental cell line, HCT 116 pSNG-xL#2 and 3 showed 50% decrease in Bcl-xL and stable Bcl-2. HCT 116 pSNG-xL#3 showed a concomitant decrease (50%) in Bcl-2. A decrease in Bcl-xL sensitised cells to the small molecule inhibitor of Bcl-xL, Antimycin A3 and the DNA topoisomerase I inhibitors, SN-38 and camptothecin, but not to doxorubicin. HCT 116 pSNG-xL#1 produced a moderate increase in both senescence and apoptosis and a limited increase in SN-38 induced cell death while HCT 116 pSNG-xL#2 produced an increase in apoptosis but reduced senescence. Finally, when both Bcl-xL and Bcl-2 were decreased to a similar degree (HCT 116 pSNG-xL#3), senescence was significantly increased but apoptosis was limited. This effect was confirmed in vivo after administration of irinotecan and was associated with greater anti-tumour effect. Optimal growth inhibitory effect was therefore observed when both Bcl-xL and Bcl-2 were decreased to a similar extent. Antimycin A3, in combination with SN-38 recapitulated this phenotype in HCT 116 cells, suggesting a potential role for small molecule inhibitors of Bcl-xL/Bcl-2 in the treatment of colorectal cancer, potentially in combination with irinotecan.

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Acknowledgments

We are very grateful to Prof. Hockenbery for providing Antimycin A3.

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Authors and Affiliations

  1. CRUK Pharmacology and Drug Development Group, Cancer Research UK Centre, University of Edinburgh, Crewe Road, Edinburgh, EH4 2XR, UK

    S. M. Guichard, M. L. Hua, P. Kang, J. S. Macpherson & D. I. Jodrell

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  1. S. M. Guichard
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  2. M. L. Hua
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  3. P. Kang
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  4. J. S. Macpherson
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  5. D. I. Jodrell
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Correspondence to S. M. Guichard.

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Guichard, S.M., Hua, M.L., Kang, P. et al. Short hairpin RNAs targeting Bcl-xL modulate senescence and apoptosis following SN-38 and irinotecan exposure in a colon cancer model. Cancer Chemother Pharmacol 60, 651–660 (2007). https://doi.org/10.1007/s00280-006-0408-1

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  • DOI: https://doi.org/10.1007/s00280-006-0408-1

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