Myofibroblasts are responsible for collagen synthesis in the stroma of human hepatocellular carcinoma: an in vivo and in vitro study
Section snippets
Human liver tissues
Tissues from ten hepatocellular carcinomas were obtained from patients undergoing either a partial hepatectomy or a liver transplantation. Histological grading of tumors was done according to Edmondson & Steiner's criteria (18). Samples were snap frozen in iso-pentane cooled in liquid nitrogen, and stored at −80°C until use.
Immunohistochemistry
Serial 5-μm sections were cut, fixed for 10 min in acetone at room temperature and processed using an indirect three-step immunoperoxidase procedure to detect α-SMA positive
Liver specimens and clinical data
The tumors came from eight males and two females, 51–68 years old (mean: 61.0), with viral (5/10) or non-viral (5/10) liver pathology. The tumors ranged from 2 cm to 14 cm (mean: 6.4 cm) and from grade 1 to 4, grade 2 being the most frequent. The non-tumoral liver was cirrhotic in 6/10, had some kind of non-cirrhotic fibrosis in two additional cases, and was non-fibrotic in the two remaining cases.
In situ detection of MF and ECM proteins and transcripts in HCC (Fig. 1 and 2)
Immunohistochemistry revealed the presence of numerous α-SMA positive cells in HCC tissues. The
Discussion
In this study, we show, using in situ hybridization, that the synthesis of type I, IV, V and VI collagens in HCC takes place in MF and not in tumoral hepatocytes or endothelial cells. The cellular type(s) responsible for the synthesis of ECM molecules in HCC had not previously been clearly identified. Several studies, using immunohistochemistry, have suggested that tumoral hepatocytes were mainly responsible for this synthesis 3., 15.. This is in contrast to our results obtained by in situ
Acknowledgements
This work was supported by grants from Comités de la Gironde et de la Dordogne from the Ligue Nationale contre le Cancer, Association pour la Recherche sur le Cancer, Fédération des Groupements d'Entreprises Françaises pour la Lutte Contre le Cancer, and Région Aquitaine.
SF was a recipient of a fellowship from Comité de la Dordogne from the Ligue Nationale Contre Le Cancer and from the Fondation Pour La Recherche Médicale.
We thank S. Milani for invaluable help in setting up the combined
References (41)
- et al.
The extracellular matrix in hepatocellular carcinoma shows different localization patterns depending on the differentiation and the histological pattern of tumors: immunohistochemical analysis
J Hepatol
(1994) - et al.
Fine structural analysis of the human pro-α1(I) collagen gene. Promoter structure, Alu I repeats, and polymorphic transcripts
J Biol Chem
(1985) - et al.
cDNA clones coding for the pro-α1(IV) chain of human type IV procollagen reveal an unusual homology of amino acid sequences in two halves of the carboxyl-terminal domain
J Biol Chem
(1985) - et al.
Human myofibroblast-like cells obtained by outgrowth are representative of the fibrogenic cells in the liver
Hepatology
(1995) - et al.
Quantitation of sinusoid-like vessels in hepatocellular carcinoma: its clinical and prognostic significance
Hepatology
(1997) - et al.
Single step method of RNA isolation by acid guanidinium thiocyanate phenol chloroform extraction
Anal Biochem
(1987) - et al.
Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates
Anal Biochem
(1980) - et al.
Evaluation of different staining procedures for the quantification of fibroblasts cultured in 96-well plates
Anal Biochem
(1991) - et al.
Evidence that “myofibroblast-like” cells are the cellular source of capsular collagen in hepatocellular carcinoma
J Hepatol
(1997) - et al.
Human hepatic myofibroblasts increase invasiveness of hepatocellular carcinoma cells. Evidence for a role of hepatocyte growth factor
Hepatology
(1997)
Evaluation of hepatocellular carcinoma aggressiveness by a panel of extracellular matrix antigens
Am J Pathol
Differential expression of extracellular matrix proteins and integrins in hepatocellular carcinoma and chronic liver disease
Anticancer Res
Extracellular matrix composition and integrin expression in early hepatocarcinogenesis in human cirrhotic liver
J Pathol
Hepatocellular carcinoma developed on noncirrhotic livers. Sinusoids in hepatocellular carcinoma
Arch Pathol Lab Med
Modulation of alpha smooth muscle actin and desmin expression in perisinusoidal cells of normal and diseased human liver
Am J Pathol
Myofibroblasts in hepatitis B related cirrhosis and hepatocellular carcinoma
J Clin Pathol
Expression of α-smooth muscle actin in liver diseases
J Korean Med Sci
α-Smooth muscle actin positive perisinusoidal stromal cells in human hepatocellular carcinoma
Hepatology
Genes of laminin B1 chain, α1(IV) chain of type IV collagen, and 72-Kd type IV collagenase are mainly expressed by the stromal cells of lung carcinomas
Am J Pathol
Expression and in-situ localization of genes coding for extracellular matrix proteins and extracellular degrading proteases in pancreatic cancer
Int J Cancer
Cited by (94)
Stromal regulation of tumor-associated lymphatics
2020, Advanced Drug Delivery ReviewsCitation Excerpt :Within a tumor, CAFs have a pivotal role with respect to formation and remodeling of the mechanical environment. Here they serve as the dominant source of ECM, producing amongst others, collagen family members, tenascin C, laminins, versican, fibronectin, hyaluronic acid, proteoglycans, glycosaminoglycans and MMPs [159–163]. Increased ECM reorganization is catalyzed by CAF-derived MMPs and lysyl oxidase (LOX) family members which cross-link collagens to mediate fiber elongation and fiber realignment, and promote a stiffer medium conducive with disease progression [164–167].
Breast cancer-derived extracellular vesicles stimulate myofibroblast differentiation and pro-angiogenic behavior of adipose stem cells
2017, Matrix BiologyCitation Excerpt :Excessive fibrotic remodeling of the stroma, termed desmoplasia, is a hallmark of breast cancer that is mediated by myofibroblasts and correlates with an advanced, invasive phenotype and worse clinical prognosis [1–3]. Myofibroblasts are highly contractile cells that contain alpha smooth muscle actin (α-SMA)-positive stress fibers [4] and are able to deposit and remodel key fibrillar components of the extracellular matrix (ECM) including type I collagen and fibronectin [5,6]. The resulting compositional, structural, and mechanical changes of the ECM directly impact tumor cell aggressiveness [7–9].
Hepatocellular carcinoma: Mouse models and the potential roles of proteases
2017, Cancer LettersPancreatic cancer stromal biology and therapy
2015, Genes and DiseasesCitation Excerpt :Generation of stromal cells via epithelial-to-mesenchymal transition. Myofibroblasts are the most abundant stromal cells and are actively involved in the development of pancreatic cancer stroma.64,72–76 However, recent discoveries that myofibroblasts can be derived from epithelial cells have provided a new impetus for investigating the processes involved in myofibroblast formation in the fibrotic and malignant contexts.76–84
Endothelial cell plasticity in kidney fibrosis and disease
2023, Acta PhysiologicaNitric Oxide Derived from Cytoglobin-Deficient Hepatic Stellate Cells Causes Suppression of Cytochrome c Oxidase Activity in Hepatocytes
2023, Antioxidants and Redox Signaling