Transforming growth factor-beta1 induces microvascular abnormalities through a down-modulation of neural cell adhesion molecule in human hepatocellular carcinoma

Piera Balzarini, Anna Benetti, Gloria Invernici, Silvia Cristini, Sonia Zicari, Arnaldo Caruso, Luisa B. Gatta, Angiola Berenzi, Luisa Imberti, Cinzia Zanotti, Nazario Portolani, Stefano M. Giulini, Maura Ferrari, Emilio Ciusani, Stefania E. Navone, Alessandra Canazza, Eugenio A. Parati, Giulio Alessandri

Research output: Contribution to journalArticle

Abstract

Hepatocellular carcinoma (HCC) is a very angiogenic and malignant cancer. Conventional chemotherapy is poorly effective because of the abnormal structural organization of HCC-infiltrating vessels. In previous work, we demonstrated that HCC angiogenesis is driven by transforming growth factor beta-1(TGF-Β1)/CD105 axis, stimulating liver-derived microvascular endothelial cells (Ld-MECs) migration. As TGF-Β1 also affects mural cells (MCs) recruitment and maturation, we asked whether it may contribute to HCC-induced vascular abnormalities. HCC and adjacent non-neoplastic liver (nNL) biopsies obtained from 12 patients were analyzed by immunohistochemistry for angiogenic markers CD105, TGF-Β1, CD44 and vascular endothelial growth factor-a (VEGFa) and for MC markers NG2, α-smooth muscle actin (αSMA) and neural cell adhesion molecule (NCAM). The same markers were also investigated by immunocytochemistry on cultured HCC-derived stromal cells (HCC-StCs) and nNL-derived StCs (nNL-StCs) isolated from the same liver biopsies. Angiogenic factors released by StCs were analyzed by ELISA and the interaction between StCs and Ld-MECs by adhesion assay. Compared with nNL, HCC biopsies showed increased angiogenic markers and αSMA that was localized in vessels. By contrast, NG2 and NCAM were substantially localized in tumor cells but absent in vessels and stroma. Cultured HCC-StCs showed less expression of NG2, αSMA and NCAM. They also demonstrated a lower capacity to release angiogenic factors and adhered on Ld-MECs. HCC-StCs and nNL-StCs treated with TGF-Β1 or with of HepG2 (a human hepatoma cell line) derived conditioned medium (CM), down-modulated NCAM expression, whereas anti-NCAM antibodies significantly reduced the adhesion of StCs to Ld-MECs. By further blocking TGF-Β1 with anti-TGF-Β1 antibodies or with Ly-364947 (a specific inhibitor TGF-Β1-receptor) adhesion to Ld-MECs and NCAM expression respectively was partially restored. TGF-Β1 contributes to HCC-induced vascular alterations by affecting the interaction between HCC-StCs and Ld-MECs through a down-modulation of NCAM expression.

Original languageEnglish
Pages (from-to)1297-1309
Number of pages13
JournalLaboratory Investigation
Volume92
Issue number9
DOIs
Publication statusPublished - Sep 2012

    Fingerprint

Keywords

  • adhesion molecules
  • angiogenesis
  • HCC
  • NCAM
  • stromal cells
  • TGF-β1
  • vascular abnormalities

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Cell Biology
  • Molecular Biology

Cite this

Balzarini, P., Benetti, A., Invernici, G., Cristini, S., Zicari, S., Caruso, A., Gatta, L. B., Berenzi, A., Imberti, L., Zanotti, C., Portolani, N., Giulini, S. M., Ferrari, M., Ciusani, E., Navone, S. E., Canazza, A., Parati, E. A., & Alessandri, G. (2012). Transforming growth factor-beta1 induces microvascular abnormalities through a down-modulation of neural cell adhesion molecule in human hepatocellular carcinoma. Laboratory Investigation, 92(9), 1297-1309. https://doi.org/10.1038/labinvest.2012.94