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Article

CTCF Expression is Essential for Somatic Cell Viability and Protection Against Cancer

1
Gene and Stem Cell Therapy Program Centenary Institute, The University of Sydney, Camperdown, 2050 NSW, Australia
2
Department of Pathology, University of Washington, 98195 Seattle, USA
3
Laboratory of Immunogenetics, Molecular Pathology Section, National Institute of Allergy & Infectious Diseases, 20852-8152 Rockville, USA
4
Cell and Molecular Therapies, Royal Prince Alfred Hospital, Camperdown, 2050 NSW, Australia
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2018, 19(12), 3832; https://doi.org/10.3390/ijms19123832
Received: 19 October 2018 / Revised: 11 November 2018 / Accepted: 11 November 2018 / Published: 30 November 2018
(This article belongs to the Special Issue Molecular Research of Endometrial Pathophysiology)
CCCTC-binding factor (CTCF) is a conserved transcription factor that performs diverse roles in transcriptional regulation and chromatin architecture. Cancer genome sequencing reveals diverse acquired mutations in CTCF, which we have shown functions as a tumour suppressor gene. While CTCF is essential for embryonic development, little is known of its absolute requirement in somatic cells and the consequences of CTCF haploinsufficiency. We examined the consequences of CTCF depletion in immortalised human and mouse cells using shRNA knockdown and CRISPR/Cas9 genome editing as well as examined the growth and development of heterozygous Ctcf (Ctcf+/−) mice. We also analysed the impact of CTCF haploinsufficiency by examining gene expression changes in CTCF-altered endometrial carcinoma. Knockdown and CRISPR/Cas9-mediated editing of CTCF reduced the cellular growth and colony-forming ability of K562 cells. CTCF knockdown also induced cell cycle arrest and a pro-survival response to apoptotic insult. However, in p53 shRNA-immortalised Ctcf+/− MEFs we observed the opposite: increased cellular proliferation, colony formation, cell cycle progression, and decreased survival after apoptotic insult compared to wild-type MEFs. CRISPR/Cas9-mediated targeting in Ctcf+/− MEFs revealed a predominance of in-frame microdeletions in Ctcf in surviving clones, however protein expression could not be ablated. Examination of CTCF mutations in endometrial cancers showed locus-specific alterations in gene expression due to CTCF haploinsufficiency, in concert with downregulation of tumour suppressor genes and upregulation of estrogen-responsive genes. Depletion of CTCF expression imparts a dramatic negative effect on normal cell function. However, CTCF haploinsufficiency can have growth-promoting effects consistent with known cancer hallmarks in the presence of additional genetic hits. Our results confirm the absolute requirement for CTCF expression in somatic cells and provide definitive evidence of CTCF’s role as a haploinsufficient tumour suppressor gene. CTCF genetic alterations in endometrial cancer indicate that gene dysregulation is a likely consequence of CTCF loss, contributing to, but not solely driving cancer growth. View Full-Text
Keywords: CTCF; tumour suppressor gene; haploinsufficiency; zinc finger; CRISPR/Cas9; cancer; endometrial cancer; gene editing CTCF; tumour suppressor gene; haploinsufficiency; zinc finger; CRISPR/Cas9; cancer; endometrial cancer; gene editing
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MDPI and ACS Style

Bailey, C.G.; Metierre, C.; Feng, Y.; Baidya, K.; Filippova, G.N.; Loukinov, D.I.; Lobanenkov, V.V.; Semaan, C.; Rasko, J.E. CTCF Expression is Essential for Somatic Cell Viability and Protection Against Cancer. Int. J. Mol. Sci. 2018, 19, 3832. https://doi.org/10.3390/ijms19123832

AMA Style

Bailey CG, Metierre C, Feng Y, Baidya K, Filippova GN, Loukinov DI, Lobanenkov VV, Semaan C, Rasko JE. CTCF Expression is Essential for Somatic Cell Viability and Protection Against Cancer. International Journal of Molecular Sciences. 2018; 19(12):3832. https://doi.org/10.3390/ijms19123832

Chicago/Turabian Style

Bailey, Charles G., Cynthia Metierre, Yue Feng, Kinsha Baidya, Galina N. Filippova, Dmitri I. Loukinov, Victor V. Lobanenkov, Crystal Semaan, and John E. Rasko. 2018. "CTCF Expression is Essential for Somatic Cell Viability and Protection Against Cancer" International Journal of Molecular Sciences 19, no. 12: 3832. https://doi.org/10.3390/ijms19123832

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