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Functional relationship between CFTR and RAC3 expression for maintaining cancer cell stemness in human colorectal cancer

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Abstract

Purpose

CFTR mutations not only cause cystic fibrosis, but also increase the risk of colorectal cancer. A putative role of CFTR in colorectal cancer patients without cystic fibrosis has so far, however, not been investigated. RAC3 is a nuclear receptor coactivator that has been found to be overexpressed in several human tumors, and to be required for maintaining cancer stemness. Here, we investigated the functional relationship between CFTR and RAC3 for maintaining cancer stemness in human colorectal cancer.

Methods

Cancer stemness was investigated by analysing the expression of stem cell markers, clonogenic growth and selective retention of fluorochrome, using stable transfection of shCFTR or shRAC3 in HCT116 colorectal cancer cells. In addition, we performed pathway enrichment and network analyses in both primary human colorectal cancer samples (TCGA, Xena platform) and Caco-2 colorectal cancer cells including (1) CD133+ or CD133- side populations and (2) CFTRwt or CFTRmut cells (ConsensusPathDB, STRING, Cytoscape, GeneMANIA).

Results

We found that the CD133+ side population expresses higher levels of RAC3 and CFTR than the CD133- side population. RAC3 overexpression increased CFTR expression, whereas CFTR downregulation inhibited the cancer stem phenotype. CFTR mRNA levels were found to be increased in colorectal cancer samples from patients without cystic fibrosis compared to those with CFTR mutations, and this correlated with an increased expression of RAC3. The expression pattern of a gene set involved in inflammatory response and nuclear receptor modulation in CD133+ Caco-2 cells was found to be shared with that in CFTRwt Caco-2 cells. These genes may contribute to colorectal cancer development.

Conclusions

CFTR may play a non-tumor suppressor role in colorectal cancer development and maintenance involving enhancement of the expression of a set of genes related to cancer stemness and development in patients without CFTR mutations.

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Abbreviations

ABC transporters:

ATP-Binding Cassette transporters

ATP:

Adenosine Triphosphate

BSA:

Bovine Serum Albumin

CFTR:

Cystic Fibrosis Transmembrane Conductance Regulator

EGF:

Epidermal Growth Factor

MDR:

Multiple Drug Resistance

NF-ҡB:

Nuclear Factor Kappa B

OCT4:

Octamer-Binding Transcription Factor 4

PBS:

Phosphate Buffered Saline

PPI:

Protein-Protein Interaction

shCFTR:

Short Hairpin for CFTR

shRAC3:

Short Hairpin for RAC3

TNF:

Tumor Necrosis Factor

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Funding

This work was supported by Consejo Nacional de Investigaciones Científicas y Técnicas [PIP 11220150100118CO] and Agencia Nacional de Promoción Científica y Tecnológica [PICT 1424, PBID 2014].

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Authors and Affiliations

  1. Laboratorio de Biología Molecular y Apoptosis, Instituto de Investigaciones Médicas Alfredo Lanari, IDIM-UBA-CONICET, Facultad de Medicina, Universidad de Buenos Aires, Combatientes de Malvinas 3150. Cuerpo II, Piso 1, C1427ARO, Buenos Aires, Argentina

    Alejandra Graciela Palma, Mileni Soares Machado, María Cecilia Lira, Francisco Rosa, María Fernanda Rubio & Mónica Alejandra Costas

  2. CONICET, Buenos Aires, Argentina

    María Fernanda Rubio, Gabriela Marino, Basilio Aristidis Kotsias & Mónica Alejandra Costas

  3. Laboratorio de Canales Iónicos, Instituto de Investigaciones Médicas Alfredo Lanari, IDIM-UBA-CONICET, Facultad de Medicina, Universidad de Buenos Aires, Combatientes de Malvinas 3150, C1427ARO, Buenos Aires, Argentina

    Gabriela Marino & Basilio Aristidis Kotsias

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  1. Alejandra Graciela Palma
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  2. Mileni Soares Machado
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  3. María Cecilia Lira
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  4. Francisco Rosa
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  5. María Fernanda Rubio
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  6. Gabriela Marino
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  7. Basilio Aristidis Kotsias
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  8. Mónica Alejandra Costas
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Contributions

Alejandra Graciela Palma: methodology, investigation, visualization, formal analysis; Mileni Soares Machado: methodology, investigation, visualization; María Cecilia Lira: investigation, formal analysis; Francisco Rosa: investigation, formal analysis; María Fernanda Rubio: Review and editing, visualization; Gabriela Marino: formal analysis, visualization; Basilio Aristidis Kotsias: conceptualization, writing, review and editing; Mónica Alejandra Costas: supervision, conceptualization, funding acquisition, project administration, original draft writing, formal analysis, visualization.

Corresponding author

Correspondence to Mónica Alejandra Costas.

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Palma, A.G., Soares Machado, M., Lira, M.C. et al. Functional relationship between CFTR and RAC3 expression for maintaining cancer cell stemness in human colorectal cancer. Cell Oncol. 44, 627–641 (2021). https://doi.org/10.1007/s13402-021-00589-x

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