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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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Angiogenin serum levels are not increased in ALD.Angiogenin serum levels were measured in control individuals (n = 5) and patients with liver pathologies: ALD, alcoholic liver disease (n = 9, age: 48.1 ± 3.8), HC, hepatic cirrhosis (n = 7, age: 58.1 ± 2.4, MELD: 10.3 ± 1.6), and HCD, hepatic cirrhosis decompensated (n = 9 age: 51.3 ± 2.2, MELD: 14.3 ± 1.8). Additional data of patients is provided in Suppl. Fig. 2.
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f6: Angiogenin serum levels are not increased in ALD.Angiogenin serum levels were measured in control individuals (n = 5) and patients with liver pathologies: ALD, alcoholic liver disease (n = 9, age: 48.1 ± 3.8), HC, hepatic cirrhosis (n = 7, age: 58.1 ± 2.4, MELD: 10.3 ± 1.6), and HCD, hepatic cirrhosis decompensated (n = 9 age: 51.3 ± 2.2, MELD: 14.3 ± 1.8). Additional data of patients is provided in Suppl. Fig. 2.

Mentions: HSC activation is a hallmark of progressive fibrosis during hepatic deterioration in liver diseases. Our data indicates that angiogenin is an inducer of HSC activation in vitro, however, a relevant role of angiogenin in the HSC activation during liver fibrosis has not been previously proposed. To evaluate the contribution of angiogenin on HSC activation during chronic liver injury, we analyzed the levels of angiogenin in serum samples from individuals with different stages of alcoholic liver disease (ALD). No differences in angiogenin serum content were observed in alcoholic patients with fibrosis or compensated cirrhosis compared to control individuals (Fig. 6). Of note, cirrhotic decompensated patients displayed an important reduction of angiogenin levels, in line with previous results14. Therefore, although angiogenin secreted by hepatoma cells, might modify liver cancer microenvironment via HSC-induced matrix remodeling, and angiogenin serum levels have been found increase in HCC patients as the tumor vascularity increase14, angiogenin concentration is not a reliable pre-neoplasic marker for liver cancer development, at least in a background of ALD. Therefore, angiogenin induced by hepatoma cells may induce further activation of already active HSCs, as observed in LX2 cells, contributing to HCC development, but without playing a role in initial HSC activation during liver fibrosis.


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)

Angiogenin serum levels are not increased in ALD.Angiogenin serum levels were measured in control individuals (n = 5) and patients with liver pathologies: ALD, alcoholic liver disease (n = 9, age: 48.1 ± 3.8), HC, hepatic cirrhosis (n = 7, age: 58.1 ± 2.4, MELD: 10.3 ± 1.6), and HCD, hepatic cirrhosis decompensated (n = 9 age: 51.3 ± 2.2, MELD: 14.3 ± 1.8). Additional data of patients is provided in Suppl. Fig. 2.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f6: Angiogenin serum levels are not increased in ALD.Angiogenin serum levels were measured in control individuals (n = 5) and patients with liver pathologies: ALD, alcoholic liver disease (n = 9, age: 48.1 ± 3.8), HC, hepatic cirrhosis (n = 7, age: 58.1 ± 2.4, MELD: 10.3 ± 1.6), and HCD, hepatic cirrhosis decompensated (n = 9 age: 51.3 ± 2.2, MELD: 14.3 ± 1.8). Additional data of patients is provided in Suppl. Fig. 2.
Mentions: HSC activation is a hallmark of progressive fibrosis during hepatic deterioration in liver diseases. Our data indicates that angiogenin is an inducer of HSC activation in vitro, however, a relevant role of angiogenin in the HSC activation during liver fibrosis has not been previously proposed. To evaluate the contribution of angiogenin on HSC activation during chronic liver injury, we analyzed the levels of angiogenin in serum samples from individuals with different stages of alcoholic liver disease (ALD). No differences in angiogenin serum content were observed in alcoholic patients with fibrosis or compensated cirrhosis compared to control individuals (Fig. 6). Of note, cirrhotic decompensated patients displayed an important reduction of angiogenin levels, in line with previous results14. Therefore, although angiogenin secreted by hepatoma cells, might modify liver cancer microenvironment via HSC-induced matrix remodeling, and angiogenin serum levels have been found increase in HCC patients as the tumor vascularity increase14, angiogenin concentration is not a reliable pre-neoplasic marker for liver cancer development, at least in a background of ALD. Therefore, angiogenin induced by hepatoma cells may induce further activation of already active HSCs, as observed in LX2 cells, contributing to HCC development, but without playing a role in initial HSC activation during liver fibrosis.

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