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Angiogenin secretion from hepatoma cells activates hepatic stellate cells to amplify a self-sustained cycle promoting liver cancer.

Bárcena C, Stefanovic M, Tutusaus A, Martinez-Nieto GA, Martinez L, García-Ruiz C, de Mingo A, Caballeria J, Fernandez-Checa JC, Marí M, Morales A - Sci Rep (2015)

Bottom Line: Protein microarray secretome analyses revealed angiogenin as the most robust and selective protein released by HCC compared to LX2 secreted molecules.Moreover, angiogenin antagonism by blocking antibodies or angiogenin inhibitor neomycin decreased in vitro HSC activation by conditioned media or recombinant angiogenin.These findings indicate that targeting angiogenin signaling may be of potential relevance in HCC management.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Death and Proliferation, IIBB-CSIC, IDIBAPS, Barcelona, Spain.

ABSTRACT
Hepatocellular carcinoma (HCC) frequently develops in a pro-inflammatory and pro-fibrogenic environment with hepatic stellate cells (HSCs) remodeling the extracellular matrix composition. Molecules secreted by liver tumors contributing to HSC activation and peritumoral stromal transformation remain to be fully identified. Here we show that conditioned medium from HCC cell lines, Hep3B and HepG2, induced primary mouse HSCs transdifferentiation, characterized by profibrotic properties and collagen modification, with similar results seen in the human HSC cell line LX2. Moreover, tumor growth was enhanced by coinjection of HepG2/LX2 cells in a xenograft murine model, supporting a HCC-HSC crosstalk in liver tumor progression. Protein microarray secretome analyses revealed angiogenin as the most robust and selective protein released by HCC compared to LX2 secreted molecules. In fact, recombinant angiogenin induced in vitro HSC activation requiring its nuclear translocation and rRNA transcriptional stimulation. Moreover, angiogenin antagonism by blocking antibodies or angiogenin inhibitor neomycin decreased in vitro HSC activation by conditioned media or recombinant angiogenin. Finally, neomycin administration reduced tumor growth of HepG2-LX2 cells coinjected in mice. In conclusion, angiogenin secretion by HCCs favors tumor development by inducing HSC activation and ECM remodeling. These findings indicate that targeting angiogenin signaling may be of potential relevance in HCC management.

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Schematic representation of angiogenin role in HCC/HSC crosstalk.Angiogenin secreted from hepatoma cells is an inducer of HSC transformation by changing numerous proteins involved in ECM remodeling. Among them, specific fibrillar components such as collagen, type I, alpha 1, encoded by the COL1A1 gene, are induced, as well as abnormal expression of enzymes that degrade type IV and V collagens and other extracellular matrix proteins, such us MMP9, tissue inhibitors of metalloproteinases (TIMP) or cysteine proteinases such as cathepsin B. Consequently, physiologic ECM formation is altered due to HSC induction, providing a profibrogenic environment that facilitates tumor growth. In summary, liver tumor promotes its own development via angiogenin-dependent HSC activation, and antagonism of angiogenin signaling, as neomycin does, may be an interesting approach to halter liver cancer progression.
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f8: Schematic representation of angiogenin role in HCC/HSC crosstalk.Angiogenin secreted from hepatoma cells is an inducer of HSC transformation by changing numerous proteins involved in ECM remodeling. Among them, specific fibrillar components such as collagen, type I, alpha 1, encoded by the COL1A1 gene, are induced, as well as abnormal expression of enzymes that degrade type IV and V collagens and other extracellular matrix proteins, such us MMP9, tissue inhibitors of metalloproteinases (TIMP) or cysteine proteinases such as cathepsin B. Consequently, physiologic ECM formation is altered due to HSC induction, providing a profibrogenic environment that facilitates tumor growth. In summary, liver tumor promotes its own development via angiogenin-dependent HSC activation, and antagonism of angiogenin signaling, as neomycin does, may be an interesting approach to halter liver cancer progression.

Mentions: HCC onset and progression is a complex process that requires multifactorial players23. Liver injury associated with inflammation and fibrosis precedes the occurrence of HCC, suggesting that HSC activation orchestrates a favorable scenario for HCC growth. Previous observations using a co-culture approach between HCC cells and primary HSC have suggested that activated HSCs play a supportive role in tumor progression by stimulating pro-angiogenic events and promoting tumor survival2728. The role of disease-specific proteins in the cell secretome is crucial in regulating cell-to-cell and cell-to-extracellular matrix interactions, and the identification of specific molecules present in conditioned media from cell lines is an interesting approach for biomedical research29. Comparing the protein content of two well-studied hepatoma cell lines (HepG2 and Hep3B) to the secretome composition of a human myofibroblastic cell line (LX2) has obvious limitations, but may provide hints of molecules involved in the cellular cross-talk during liver cancer implantation and progression. By doing so, our data identify angiogenin as novel player in a positive loop by which HCC mediates HSC activation, which accelerates and amplifies HCC development (Figure 8). Our findings indicate that HCC-mediated secretion of angiogenin induces, or even aggravate, HSC activation promoting a phenotype with ECM remodeling activity. On the contrary, angiogenin antagonism may affect cancer microenvironment, reducing vessel formation and slowing down tumor growth in in vivo HCC models.


Angiogenin secretion from hepatoma cells activates hepatic stellate cells to amplify a self-sustained cycle promoting liver cancer.

Bárcena C, Stefanovic M, Tutusaus A, Martinez-Nieto GA, Martinez L, García-Ruiz C, de Mingo A, Caballeria J, Fernandez-Checa JC, Marí M, Morales A - Sci Rep (2015)

Schematic representation of angiogenin role in HCC/HSC crosstalk.Angiogenin secreted from hepatoma cells is an inducer of HSC transformation by changing numerous proteins involved in ECM remodeling. Among them, specific fibrillar components such as collagen, type I, alpha 1, encoded by the COL1A1 gene, are induced, as well as abnormal expression of enzymes that degrade type IV and V collagens and other extracellular matrix proteins, such us MMP9, tissue inhibitors of metalloproteinases (TIMP) or cysteine proteinases such as cathepsin B. Consequently, physiologic ECM formation is altered due to HSC induction, providing a profibrogenic environment that facilitates tumor growth. In summary, liver tumor promotes its own development via angiogenin-dependent HSC activation, and antagonism of angiogenin signaling, as neomycin does, may be an interesting approach to halter liver cancer progression.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4300499&req=5

f8: Schematic representation of angiogenin role in HCC/HSC crosstalk.Angiogenin secreted from hepatoma cells is an inducer of HSC transformation by changing numerous proteins involved in ECM remodeling. Among them, specific fibrillar components such as collagen, type I, alpha 1, encoded by the COL1A1 gene, are induced, as well as abnormal expression of enzymes that degrade type IV and V collagens and other extracellular matrix proteins, such us MMP9, tissue inhibitors of metalloproteinases (TIMP) or cysteine proteinases such as cathepsin B. Consequently, physiologic ECM formation is altered due to HSC induction, providing a profibrogenic environment that facilitates tumor growth. In summary, liver tumor promotes its own development via angiogenin-dependent HSC activation, and antagonism of angiogenin signaling, as neomycin does, may be an interesting approach to halter liver cancer progression.
Mentions: HCC onset and progression is a complex process that requires multifactorial players23. Liver injury associated with inflammation and fibrosis precedes the occurrence of HCC, suggesting that HSC activation orchestrates a favorable scenario for HCC growth. Previous observations using a co-culture approach between HCC cells and primary HSC have suggested that activated HSCs play a supportive role in tumor progression by stimulating pro-angiogenic events and promoting tumor survival2728. The role of disease-specific proteins in the cell secretome is crucial in regulating cell-to-cell and cell-to-extracellular matrix interactions, and the identification of specific molecules present in conditioned media from cell lines is an interesting approach for biomedical research29. Comparing the protein content of two well-studied hepatoma cell lines (HepG2 and Hep3B) to the secretome composition of a human myofibroblastic cell line (LX2) has obvious limitations, but may provide hints of molecules involved in the cellular cross-talk during liver cancer implantation and progression. By doing so, our data identify angiogenin as novel player in a positive loop by which HCC mediates HSC activation, which accelerates and amplifies HCC development (Figure 8). Our findings indicate that HCC-mediated secretion of angiogenin induces, or even aggravate, HSC activation promoting a phenotype with ECM remodeling activity. On the contrary, angiogenin antagonism may affect cancer microenvironment, reducing vessel formation and slowing down tumor growth in in vivo HCC models.

Bottom Line: Protein microarray secretome analyses revealed angiogenin as the most robust and selective protein released by HCC compared to LX2 secreted molecules.Moreover, angiogenin antagonism by blocking antibodies or angiogenin inhibitor neomycin decreased in vitro HSC activation by conditioned media or recombinant angiogenin.These findings indicate that targeting angiogenin signaling may be of potential relevance in HCC management.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Death and Proliferation, IIBB-CSIC, IDIBAPS, Barcelona, Spain.

ABSTRACT
Hepatocellular carcinoma (HCC) frequently develops in a pro-inflammatory and pro-fibrogenic environment with hepatic stellate cells (HSCs) remodeling the extracellular matrix composition. Molecules secreted by liver tumors contributing to HSC activation and peritumoral stromal transformation remain to be fully identified. Here we show that conditioned medium from HCC cell lines, Hep3B and HepG2, induced primary mouse HSCs transdifferentiation, characterized by profibrotic properties and collagen modification, with similar results seen in the human HSC cell line LX2. Moreover, tumor growth was enhanced by coinjection of HepG2/LX2 cells in a xenograft murine model, supporting a HCC-HSC crosstalk in liver tumor progression. Protein microarray secretome analyses revealed angiogenin as the most robust and selective protein released by HCC compared to LX2 secreted molecules. In fact, recombinant angiogenin induced in vitro HSC activation requiring its nuclear translocation and rRNA transcriptional stimulation. Moreover, angiogenin antagonism by blocking antibodies or angiogenin inhibitor neomycin decreased in vitro HSC activation by conditioned media or recombinant angiogenin. Finally, neomycin administration reduced tumor growth of HepG2-LX2 cells coinjected in mice. In conclusion, angiogenin secretion by HCCs favors tumor development by inducing HSC activation and ECM remodeling. These findings indicate that targeting angiogenin signaling may be of potential relevance in HCC management.

Show MeSH
Related in: MedlinePlus