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KRAS phosphorylation regulates cell polarization and tumorigenic properties in colorectal cancer

Oncogene volume 40, pages 5730–5740 (2021)Cite this article

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Abstract

Oncogenic mutations of KRAS are found in the most aggressive human tumors, including colorectal cancer. It has been suggested that oncogenic KRAS phosphorylation at Ser181 modulates its activity and favors cell transformation. Using nonphosphorylatable (S181A), phosphomimetic (S181D), and phospho-/dephosphorylatable (S181) oncogenic KRAS mutants, we analyzed the role of this phosphorylation to the maintenance of tumorigenic properties of colorectal cancer cells. Our data show that the presence of phospho-/dephosphorylatable oncogenic KRAS is required for preserving the epithelial organization of colorectal cancer cells in 3D cultures, and for supporting subcutaneous tumor growth in mice. Interestingly, gene expression differed according to the phosphorylation status of KRAS. In DLD-1 cells, CTNNA1 was only expressed in phospho-/dephosphorylatable oncogenic KRAS-expressing cells, correlating with cell polarization. Moreover, lack of oncogenic KRAS phosphorylation leads to changes in expression of genes related to cell invasion, such as SERPINE1, PRSS1,2,3, and NEO1, and expression of phosphomimetic oncogenic KRAS resulted in diminished expression of genes involved in enterocyte differentiation, such as HNF4G. Finally, the analysis, in a public data set of human colorectal cancer, of the gene expression signatures associated with phosphomimetic and nonphosphorylatable oncogenic KRAS suggests that this post-translational modification regulates tumor progression in patients.

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Fig. 1: Stable expression of oncogenic KRAS phosphomutants induces differential cell morphology.
Fig. 2: Status of oncogenic KRAS phosphorylation at Ser181 has an impact on gene expression.
Fig. 3: Oncogenic KRAS phosphorylation/dephosphorylation cycle at Ser181 is necessary to induce an epithelial polarized structure.
Fig. 4: Oncogenic KRAS phosphorylation/dephosphorylation cycle at Ser181 regulates cell-invasive capacity.
Fig. 5: Phosphorylation at Ser181 of oncogenic KRAS is necessary for tumor growth.
Fig. 6: Expression of S181D and S181A signature in human CRC primary tumors and normal colon.

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Data availability

The datasets generated during the current study are available in the GEO database repository: https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE176276

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Acknowledgements

We thank the personnel of the Advanced Microscopy Unit of CCiT-UB (Campus Clinic) for help in setting up image acquisition and analysis. This work was supported by grants from the Ministerio de Economia y Competitividad and co funded by FEDER funds—a way to build Europe− (SAF2016-76239-R and PID2019-105483RB-I00 to NA, SAF2015-68016-R to GC), CIBERONC and the Government of Catalonia (grants 2017SGR1282 and PERIS SLT002/16/0037), and from Instituto de Salud Carlos III (grant PI17/01304 to MC); a FPU fellowship from Ministerio de Educación, Cultura y Deporte for DC, a FI fellowship from Generalitat de Catalunya for NP, and a Predoc-UB from University of Barcelona to BA; CR is supported by the Serra Húnter Program (Generalitat de Catalunya).

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Author notes

  1. Núria Gendrau-Sanclemente

    Present address: Program Against Cancer Therapeutic Resistance (ProCURE), Catalan Institute of Oncology, Hospital Duran i Reynals, Institut d’Investigació Biomèdica de Bellvitge (IDIBELL), L’Hospitalet de Llobregat, Barcelona, Spain

  2. Carles Barceló

    Present address: Institut d’Investigació Sanitària Illes Balears (IdISBa), Palma de Mallorca, Spain

Authors and Affiliations

  1. Department Biomedicina, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, Barcelona, Spain

    Débora Cabot, Sònia Brun, Noelia Paco, Núria Gendrau-Sanclemente, Baraa Abuasaker, Triana Ruiz-Fariña, Carles Barceló, Marta Bosch, Carles Rentero, Josep M. Estanyol, Montserrat Jaumot & Neus Agell

  2. Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain

    Débora Cabot, Sònia Brun, Noelia Paco, Baraa Abuasaker, Carles Barceló, Miriam Cuatrecasas, Marta Bosch, Carles Rentero, Montserrat Jaumot & Neus Agell

  3. Hereditary Cancer Program, Translational Research Laboratory, Catalan Institute of Oncology, ICO-IDIBELL, Hospitalet de Llobregat, Barcelona, Spain and Centro de Investigación Biomédica en Red en Cáncer (CIBERONC), Madrid, Spain

    Mireia M. Ginesta & Gabriel Capellà

  4. Departament de Fonaments Clínics, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona; Pathology Department and Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd) and Tumor Bank-Biobank, Hospital Clínic, Barcelona, Spain

    Miriam Cuatrecasas

  5. Departament de Ciències Fisiològiques, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona and Institut d’Investigació Biomèdica de Bellvitge (IDIBELL), L’Hospitalet de Llobregat, Barcelona, Spain

    Gabriel Pons

  6. Proteomics Unit, CCiT-UB, Universitat de Barcelona, Barcelona, Spain

    Josep M. Estanyol

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  1. Débora Cabot
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Contributions

DC and SB contributed equally to this work; DC, SB, NP, MMG, BA, and NG-S conducted the experiments and data analysis; CB, MC, JME, MB, CR, GC, MJ, and NA conducted data analysis and interpretation; TR-F, GP, and CR provided technical set-up and support. MJ and NA designed the study; MJ, DC, and NA wrote the paper. All authors read and approve the final paper.

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Correspondence to Montserrat Jaumot or Neus Agell.

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Cabot, D., Brun, S., Paco, N. et al. KRAS phosphorylation regulates cell polarization and tumorigenic properties in colorectal cancer. Oncogene 40, 5730–5740 (2021). https://doi.org/10.1038/s41388-021-01967-3

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