Chromosomally Stable Colon Cancer Cell Models

The cellular models consisted
of chromosomally stable human colon cancer DLD1 diploid (2N) and tetraploid
(4N) cells, along with tetraploid centrosome -amplified (4NCA) DLD1
cells and DLD1 HSET knockout (KO) cells. DLD1 2N and 4N cells were
generated and characterized as previously described.^{27 (link),41 ,42 (link)} In order to generate 4NCA cells,
we used dihydro-cytochalasin B (DCB) to transiently block cytokinesis
and induce tetraploidisation and centrosome amplification in DLD1
cells. Centrosome amplification in 4NCA cells is transient and therefore
they were generated from 2N cells for each run of the assay.^{27 (link)} The DLD1 HSET KO cells were knockout clones
generated by CRISPR and validated by sequencing to have a homozygous
deletion in the KIFC1 gene.

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Saint-Dizier F., Matthews T.P., Gregson A.M., Prevet H., McHardy T., Colombano G., Saville H., Rowlands M., Ewens C., McAndrew P.C., Tomlin K., Guillotin D., Mak G.W., Drosopoulos K., Poursaitidis I., Burke R., van Montfort R., Linardopoulos S, & Collins I. (2023). Discovery of 2-(3-Benzamidopropanamido)thiazole-5-carboxylate Inhibitors of the Kinesin HSET (KIFC1) and the Development of Cellular Target Engagement Probes. Journal of Medicinal Chemistry, 66(4), 2622-2645.

Publication 2023

A gene Assay Block Cells Cellular diploid Centrosome Colon cancer Crispr Cytochalasin b Diploid Human Tetraploid Transient

Corresponding Organization : Institute of Cancer Research

Other organizations : Breast Cancer Now

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Variable analysis

independent variables

Cellular models
Dihydro-cytochalasin B (DCB) treatment to induce tetraploidisation and centrosome amplification
CRISPR-mediated knockout of KIFC1 gene

dependent variables

Cellular ploidy (diploid, tetraploid, centrosome-amplified tetraploid)
Cellular phenotypes

control variables

Chromosomally stable human colon cancer DLD1 cells

controls

Positive control: DLD1 diploid (2N) and tetraploid (4N) cells
Negative control: Not explicitly mentioned

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