Tumorigenic transformation of human prostatic epithelial cell line RWPE‐1 by growth hormone‐releasing hormone (GHRH)

Muñoz-Moreno, Laura; Carmena, Marı́a J.; Prieto, Juan Carlos; Schally, Andrew V.; Bajo, Ana M.

doi:10.1002/pros.24339

Cited by 8 publications

(2 citation statements)

References 33 publications

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“…MCF10A cells are spontaneously immortalized [35], while RWPE-1 cells have been immortalized using HPV-18 [36]. Both cell lines, however, remain untransformed and maintain stable epithelial morphology and functional characteristics in-vitro [35][36][37][38][39][40][41][42]. HCMV has been found in both mammary epithelial cells [7] and prostate epithelial cells [43,44] in-vivo; therefore, MCF10A and RWPE-1 are physiologically relevant experimental infection models.…”

Section: Resultsmentioning

confidence: 99%

Subconfluent ARPE-19 Cells Display Mesenchymal Cell-State Characteristics and Behave Like Fibroblasts, Rather than Epithelial Cells, in Experimental HCMV Infection Studies

Golconda,

Andrade-Medina,

Oberstein

2023

Preprint

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Human cytomegalovirus (HCMV) has a broad cellular tropism and epithelial cells are important physiological targets during infection. The retinal pigment epithelial cell line ARPE-19 has been used to model HCMV infection in epithelial cells for decades and remains a commonly used cell-type for studying viral entry, replication, and the cellular response to infection. We previously found that ARPE-19 cells, despite being derived from an epithelial cell explant, express extremely low levels of canonical epithelial proteins, such as E-cadherin and EpCAM. Here, we perform comparative studies of ARPE-19 and additional epithelial cell lines with strong epithelial characteristics. We find that ARPE-19 cells cultured under subconfluent conditions resemble mesenchymal fibroblasts, rather than epithelial cells; consistent with previous studies showing that ARPE-19 cultures require extended periods of high confluency culture to maintain epithelial characteristics. By reanalyzing public gene expression data and using machine-learning, we find evidence that ARPE-19 cultures maintained across many labs exhibit mesenchymal characteristics and that the majority of studies employing ARPE-19 use them in a mesenchymal state. Lastly, by performing experimental HCMV infections across mesenchymal and epithelial cell lines, we find that permissivity to HCMV infection is positively correlated with mesenchymal cell state features, and negatively correlated with epithelial cell state features. Our work identifies the epithelial-mesenchymal cell state axis as a potential endogenous regulator of HCMV infection. It also raises awareness that sub-confluent ARPE-19 may not be suitable for modeling epithelial infection with HCMV or other human viruses.

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Section: Resultsmentioning

confidence: 99%

Subconfluent ARPE-19 Cells Display Mesenchymal Cell-State Characteristics and Behave Like Fibroblasts, Rather than Epithelial Cells, in Experimental HCMV Infection Studies

Golconda,

Andrade-Medina,

Oberstein

2023

Preprint

View full text Add to dashboard Cite

show abstract

“…MCF10A cells are spontaneously immortalized [ 53 ], while RWPE-1 cells have been immortalized using HPV-18 [ 54 ]. Both cell lines, however, remain untransformed and maintain stable epithelial morphology and functional characteristics in vitro [ 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 ]. HCMV has been found in both mammary epithelial cells [ 5 ] and prostate epithelial cells [ 61 , 62 ] in vivo; therefore, MCF10A and RWPE-1 are physiologically relevant experimental infection models.…”

Section: Resultsmentioning

confidence: 99%

Subconfluent ARPE-19 Cells Display Mesenchymal Cell-State Characteristics and Behave like Fibroblasts, Rather Than Epithelial Cells, in Experimental HCMV Infection Studies

Golconda,

Andrade-Medina,

Oberstein

2023

Viruses

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Human cytomegalovirus (HCMV) has a broad cellular tropism and epithelial cells are important physiological targets during infection. The retinal pigment epithelial cell line ARPE-19 has been used to model HCMV infection in epithelial cells for decades and remains a commonly used cell type for studying viral entry, replication, and the cellular response to infection. We previously found that ARPE-19 cells, despite being derived from an epithelial cell explant, express extremely low levels of canonical epithelial proteins, such as E-cadherin and EpCAM. Here, we perform comparative studies of ARPE-19 and additional epithelial cell lines with strong epithelial characteristics. We find that ARPE-19 cells cultured under subconfluent conditions resemble mesenchymal fibroblasts, rather than epithelial cells; this is consistent with previous studies showing that ARPE-19 cultures require extended periods of high confluency culture to maintain epithelial characteristics. By reanalyzing public gene expression data and using machine learning, we find evidence that ARPE-19 cultures maintained across many labs exhibit mesenchymal characteristics and that the majority of studies employing ARPE-19 use them in a mesenchymal state. Lastly, by performing experimental HCMV infections across mesenchymal and epithelial cell lines, we find that ARPE-19 cells behave like mesenchymal fibroblasts, producing logarithmic yields of cell-free infectious progeny, while cell lines with strong epithelial character exhibit an atypical infectious cycle and naturally restrict the production of cell-free progeny. Our work highlights important characteristics of the ARPE-19 cell line and suggests that subconfluent ARPE-19 cells may not be optimal for modeling epithelial infection with HCMV or other human viruses. It also suggests that HCMV biosynthesis and/or spread may occur quite differently in epithelial cells compared to mesenchymal cells. These differences could contribute to viral persistence or pathogenesis in epithelial tissues.

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Tumorigenic transformation of human prostatic epithelial cell line RWPE‐1 by growth hormone‐releasing hormone (GHRH)

et al. 2022

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Background: Growth hormone-releasing hormone (GHRH) and its receptors have been implicated in the progression of various tumors. In this study, we analyzed the carcinogenetic potential of exposure to GHRH of a nontumor human prostate epithelial cell line (RWPE-1) as well as its transforming effect in a xenograft model. Methods: We performed cell viability, cell proliferation, adhesion and migration assays. In addition, metalloprotease (MMP)−2 activity by means gelatin zymography, GHRH-R subcellular location using confocal immunofluorescence microscopy and vascular endothelial growth factor (VEGF) levels by enzyme-linked immunoassay were assessed. Besides, we developed an in vivo model in order vivo model to determine the role of GHRH on tumorigenic transformation of RWPE-1 cells. Results: In cell cultures, we observed development of a migratory phenotype consistent with the gelatinolytic activity of MMP-2, expression of VEGF, as well as E-cadherin-mediated cell-cell adhesion and increased cell motility. Treatment with 0.1 µM GHRH for 24 h significantly increased cell viability and cell proliferation. Similar effects of GHRH were seen in RWPE-1 tumors developed by subcutaneous injection of GHRH-treated cells in athymic nude mice, 49 days after inoculation.Conclusions: Thus, GHRH appears to act as a cytokine in the transformation of RWPE-1 cells by mechanisms that likely involve epithelial-mesenchymal transition, thus reinforcing the role of GHRH in tumorigenesis of prostate.

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Tumorigenic transformation of human prostatic epithelial cell line RWPE‐1 by growth hormone‐releasing hormone (GHRH)

Cited by 8 publications

References 33 publications

Subconfluent ARPE-19 Cells Display Mesenchymal Cell-State Characteristics and Behave Like Fibroblasts, Rather than Epithelial Cells, in Experimental HCMV Infection Studies

Subconfluent ARPE-19 Cells Display Mesenchymal Cell-State Characteristics and Behave Like Fibroblasts, Rather than Epithelial Cells, in Experimental HCMV Infection Studies

Subconfluent ARPE-19 Cells Display Mesenchymal Cell-State Characteristics and Behave like Fibroblasts, Rather Than Epithelial Cells, in Experimental HCMV Infection Studies

Tumorigenic transformation of human prostatic epithelial cell line RWPE‐1 by growth hormone‐releasing hormone (GHRH)

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