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Endogenous oncogenic KRAS expression increases cell proliferation and motility in near-diploid hTERT RPE-1 cells

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posted on 2024-06-26, 10:14 authored by Naushin L Hindul, Lauren R Abbott, Sumaya MD Adan, Kees StraatmanKees Straatman, Andrew M Fry, Kouji Hirota, Kayoko TanakaKayoko Tanaka
About 18% of all human cancers carry a mutation in the KRAS gene making it among the most sought-after anti-cancer targets. However, mutant KRas protein has proved remarkably undruggable. The recent approval of the first generation of RAS inhibitors therefore marks a seminal milestone in the history of cancer research. Inevitably though, it also raises the predictable challenges of limited drug efficacies and acquired resistance. Hence, new approaches that improve our understanding of the tumorigenic mechanisms of oncogenic RAS within more physiological settings continue to be essential. Here, we have employed the near-diploid human hTERT RPE-1 cells to generate isogenic cell lines in which one of the endogenous KRAS alleles carries an oncogenic KRAS mutation at glycine 12. Cells with a KRASG12V/+, KRASG12C/+, or KRASG12D/+ genotype, together with wild-type KRASG12G(WT)/+ cells, reveal that oncogenic KRAS.G12X mutations increase cell proliferation rate and cell motility and reduced focal adhesions in KRASG12V/+ cells. EGF-induced phosphorylation of ERK and AKT was comparable between KRASG12V/+, KRASG12C/+, KRASG12D/+, and KRASG12G(WT)/+ cells. Interestingly, KRASG12X/+ cells showed varying responses to distinct inhibitors with the KRASG12V/+ and KRASG12D/+ cells more sensitive to hydroxyurea and MEK inhibitors, U0126 and trametinib, but more resistant to PI3K inhibitor, PIK-90, than the KRASG12G(WT)/+ cells. A combination of low doses of hydroxyurea and U0126 showed an additive inhibition on growth rate that was greater in KRASG12V/+ than wild-type cells. Collectively, these cell lines will be a valuable resource for studying oncogenic RAS signalling and developing effective anti-KRAS reagents with minimum cytotoxicity on wild-type cells.

Funding

Institutional Strategic Support Fund

Wellcome Trust

Find out more...

BSc program at the University of Leicester to K. T. and JSPS KAKENHI (JP20H04337, JP19KK0210, and JP16H01314) to K. H.

History

Author affiliation

College of Life Sciences Molecular & Cell Biology Professional Services

Version

  • VoR (Version of Record)

Published in

Journal of Biological Chemistry

Volume

300

Issue

6

Pagination

107409

Publisher

Elsevier BV

issn

0021-9258

eissn

1083-351X

Copyright date

2024

Available date

2024-06-26

Spatial coverage

United States

Language

en

Deposited by

Dr Kayoko Tanaka

Deposit date

2024-06-22

Data Access Statement

The datasets supporting the conclusions of this article are included within the article and its additional file.

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