A functional genomics screen reveals a strong synergistic effect between docetaxel and the mitotic gene DLGAP5 that is mediated by the androgen receptor

Hewit, K., Sandilands, E., Sanchez Martinez, R., James, D., Leung, H. Y. , Bryant, D. M. , Shanks, E. and Markert, E. K. (2018) A functional genomics screen reveals a strong synergistic effect between docetaxel and the mitotic gene DLGAP5 that is mediated by the androgen receptor. Cell Death and Disease, 9(11), 1069. (doi:10.1038/s41419-018-1115-7) (PMID:30341281) (PMCID:PMC6195526)

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Abstract

Based on a molecular classification of prostate cancer using gene expression pathway signatures, we derived a set of 48 genes in critical pathways that significantly predicts clinical outcome in all tested patient cohorts. We tested these genes in a functional genomics screen in a panel of three prostate cancer cell lines (LNCaP, PC3, DU145), using RNA interference. The screen revealed several genes whose knockdown caused strong growth inhibition in all cell lines. Additionally, we tested the gene set in the presence of docetaxel to see whether any gene exhibited additive or synergistic effects with the drug. We observed a strong synergistic effect between DLGAP5 knockdown and docetaxel in the androgen-sensitive line LNCaP, but not in the two other androgen-independent lines. We then tested whether this effect was connected to androgen pathways and found that knockdown of the androgen receptor by si-RNA attenuated the synergy significantly. Similarly, androgen desensitized LNCaP-AI cells had a higher IC50 to docetaxel and did not exhibit the synergistic interaction. Short-term exposure to enzalutamide did not significantly alter the behaviour of parental LNCaP cells. An immunofluorescence analysis in LNCaP cells suggests that under the double insult of DLGAP5 knockdown and docetaxel, cells predominantly arrest in metaphase. In contrast, the knockdown of the androgen receptor by siRNA appears to assist cells to progress through metaphase in to anaphase, even in the presence of docetaxel. Our data suggest that DLGAP5 has a unique function in stabilizing spindle formation and surviving microtubule assault from docetaxel, in an androgen-regulated cell cycle system.

Item Type:Articles
Additional Information:We acknowledge the Cancer Research UK Glasgow Centre (C596/A18076) and the RNAi screening facility at the Cancer Research UK Beatson Institute (E Shanks, K Hewit, D James), core funded by Cancer Research UK (C596/A17196).
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Markert, Dr Elke and James, Mr Daniel and Bryant, Dr David and Shanks, Dr Emma and Leung, Professor Hing and Sandilands, Dr Emma and Sanchez Martinez, Mr Rafael and Hewit, Dr Kay
Authors: Hewit, K., Sandilands, E., Sanchez Martinez, R., James, D., Leung, H. Y., Bryant, D. M., Shanks, E., and Markert, E. K.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Cell Death and Disease
Publisher:Nature Publishing Group
ISSN:2041-4889
ISSN (Online):2041-4889
Copyright Holders:Copyright © 2018 The Authors
First Published:First published in Cell Death and Disease 9(11):1069
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
190874CR-UK Centre renewalKaren VousdenNot Applicable (NA)C596/A18076Institute of Cancer Sciences