Evidence of distinct tumour-propagating cell populations with different properties in primary human hepatocellular carcinoma

Federico Colombo, Francesca Baldan, Silvia Mazzucchelli, Ines Martin-Padura, Paola Marighetti, Alessandra Cattaneo, Barbara Foglieni, Marta Spreafico, Silvana Guerneri, Marco Baccarin, Francesco Bertolini, Giorgio Rossi, Vincenzo Mazzaferro, Massimiliano Cadamuro, Marco Maggioni, Luca Agnelli, Paolo Rebulla, Daniele Prati, Laura Porretti

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Background and Aims: Increasing evidence that a number of malignancies are characterised by tumour cell heterogeneity has recently been published, but there is still a lack of data concerning liver cancers. The aim of this study was to investigate and characterise tumour-propagating cell (TPC) compartments within human hepatocellular carcinoma (HCC). Methods: After long-term culture, we identified three morphologically different tumour cell populations in a single HCC specimen, and extensively characterised them by means of flow cytometry, fluorescence microscopy, karyotyping and microarray analyses, single cell cloning, and xenotransplantation in NOD/SCID/IL2Rγ-/- mice. Results: The primary cell populations (hcc-1, -2 and -3) and two clones generated by means of limiting dilutions from hcc-1 (clone-1/7 and -1/8) differently expressed a number of tumour-associated stem cell markers, including EpCAM, CD49f, CD44, CD133, CD56, Thy-1, ALDH and CK19, and also showed different doubling times, drug resistance and tumorigenic potential. Moreover, we found that ALDH expression, in combination with CD44 or Thy-1 negativity or CD56 positivity identified subpopulations with a higher clonogenic potential within hcc-1, hcc-2 and hcc-3 primary cell populations, respectively. Karyotyping revealed the clonal evolution of the cell populations and clones within the primary tumour. Importantly, the primary tumour cell population with the greatest tumorigenic potential and drug resistance showed more chromosomal alterations than the others and contained clones with epithelial and mesenchymal features. Conclusions: Individual HCCs can harbor different self-renewing tumorigenic cell types expressing a variety of morphological and phenotypical markers, karyotypic evolution and different gene expression profiles. This suggests that the models of hepatic carcinogenesis should take into account TPC heterogeneity due to intratumour clonal evolution.

Original languageEnglish
Article numbere21369
JournalPLoS One
Volume6
Issue number6
DOIs
Publication statusPublished - 2011

Fingerprint

hepatoma
Tumors
Hepatocellular Carcinoma
Cells
clones
Population
drug resistance
Neoplasms
Clone Cells
karyotyping
Clonal Evolution
Karyotyping
cells
cloning (cells)
xenotransplantation
Drug Resistance
neoplasms
liver neoplasms
fluorescence microscopy
carcinogenesis

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Evidence of distinct tumour-propagating cell populations with different properties in primary human hepatocellular carcinoma. / Colombo, Federico; Baldan, Francesca; Mazzucchelli, Silvia; Martin-Padura, Ines; Marighetti, Paola; Cattaneo, Alessandra; Foglieni, Barbara; Spreafico, Marta; Guerneri, Silvana; Baccarin, Marco; Bertolini, Francesco; Rossi, Giorgio; Mazzaferro, Vincenzo; Cadamuro, Massimiliano; Maggioni, Marco; Agnelli, Luca; Rebulla, Paolo; Prati, Daniele; Porretti, Laura.

In: PLoS One, Vol. 6, No. 6, e21369, 2011.

Research output: Contribution to journalArticle

Colombo, Federico ; Baldan, Francesca ; Mazzucchelli, Silvia ; Martin-Padura, Ines ; Marighetti, Paola ; Cattaneo, Alessandra ; Foglieni, Barbara ; Spreafico, Marta ; Guerneri, Silvana ; Baccarin, Marco ; Bertolini, Francesco ; Rossi, Giorgio ; Mazzaferro, Vincenzo ; Cadamuro, Massimiliano ; Maggioni, Marco ; Agnelli, Luca ; Rebulla, Paolo ; Prati, Daniele ; Porretti, Laura. / Evidence of distinct tumour-propagating cell populations with different properties in primary human hepatocellular carcinoma. In: PLoS One. 2011 ; Vol. 6, No. 6.
@article{7e4cb367a7f7413f83d58d01025ae6ed,
title = "Evidence of distinct tumour-propagating cell populations with different properties in primary human hepatocellular carcinoma",
abstract = "Background and Aims: Increasing evidence that a number of malignancies are characterised by tumour cell heterogeneity has recently been published, but there is still a lack of data concerning liver cancers. The aim of this study was to investigate and characterise tumour-propagating cell (TPC) compartments within human hepatocellular carcinoma (HCC). Methods: After long-term culture, we identified three morphologically different tumour cell populations in a single HCC specimen, and extensively characterised them by means of flow cytometry, fluorescence microscopy, karyotyping and microarray analyses, single cell cloning, and xenotransplantation in NOD/SCID/IL2Rγ-/- mice. Results: The primary cell populations (hcc-1, -2 and -3) and two clones generated by means of limiting dilutions from hcc-1 (clone-1/7 and -1/8) differently expressed a number of tumour-associated stem cell markers, including EpCAM, CD49f, CD44, CD133, CD56, Thy-1, ALDH and CK19, and also showed different doubling times, drug resistance and tumorigenic potential. Moreover, we found that ALDH expression, in combination with CD44 or Thy-1 negativity or CD56 positivity identified subpopulations with a higher clonogenic potential within hcc-1, hcc-2 and hcc-3 primary cell populations, respectively. Karyotyping revealed the clonal evolution of the cell populations and clones within the primary tumour. Importantly, the primary tumour cell population with the greatest tumorigenic potential and drug resistance showed more chromosomal alterations than the others and contained clones with epithelial and mesenchymal features. Conclusions: Individual HCCs can harbor different self-renewing tumorigenic cell types expressing a variety of morphological and phenotypical markers, karyotypic evolution and different gene expression profiles. This suggests that the models of hepatic carcinogenesis should take into account TPC heterogeneity due to intratumour clonal evolution.",
author = "Federico Colombo and Francesca Baldan and Silvia Mazzucchelli and Ines Martin-Padura and Paola Marighetti and Alessandra Cattaneo and Barbara Foglieni and Marta Spreafico and Silvana Guerneri and Marco Baccarin and Francesco Bertolini and Giorgio Rossi and Vincenzo Mazzaferro and Massimiliano Cadamuro and Marco Maggioni and Luca Agnelli and Paolo Rebulla and Daniele Prati and Laura Porretti",
year = "2011",
doi = "10.1371/journal.pone.0021369",
language = "English",
volume = "6",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "6",

}

TY - JOUR

T1 - Evidence of distinct tumour-propagating cell populations with different properties in primary human hepatocellular carcinoma

AU - Colombo, Federico

AU - Baldan, Francesca

AU - Mazzucchelli, Silvia

AU - Martin-Padura, Ines

AU - Marighetti, Paola

AU - Cattaneo, Alessandra

AU - Foglieni, Barbara

AU - Spreafico, Marta

AU - Guerneri, Silvana

AU - Baccarin, Marco

AU - Bertolini, Francesco

AU - Rossi, Giorgio

AU - Mazzaferro, Vincenzo

AU - Cadamuro, Massimiliano

AU - Maggioni, Marco

AU - Agnelli, Luca

AU - Rebulla, Paolo

AU - Prati, Daniele

AU - Porretti, Laura

PY - 2011

Y1 - 2011

N2 - Background and Aims: Increasing evidence that a number of malignancies are characterised by tumour cell heterogeneity has recently been published, but there is still a lack of data concerning liver cancers. The aim of this study was to investigate and characterise tumour-propagating cell (TPC) compartments within human hepatocellular carcinoma (HCC). Methods: After long-term culture, we identified three morphologically different tumour cell populations in a single HCC specimen, and extensively characterised them by means of flow cytometry, fluorescence microscopy, karyotyping and microarray analyses, single cell cloning, and xenotransplantation in NOD/SCID/IL2Rγ-/- mice. Results: The primary cell populations (hcc-1, -2 and -3) and two clones generated by means of limiting dilutions from hcc-1 (clone-1/7 and -1/8) differently expressed a number of tumour-associated stem cell markers, including EpCAM, CD49f, CD44, CD133, CD56, Thy-1, ALDH and CK19, and also showed different doubling times, drug resistance and tumorigenic potential. Moreover, we found that ALDH expression, in combination with CD44 or Thy-1 negativity or CD56 positivity identified subpopulations with a higher clonogenic potential within hcc-1, hcc-2 and hcc-3 primary cell populations, respectively. Karyotyping revealed the clonal evolution of the cell populations and clones within the primary tumour. Importantly, the primary tumour cell population with the greatest tumorigenic potential and drug resistance showed more chromosomal alterations than the others and contained clones with epithelial and mesenchymal features. Conclusions: Individual HCCs can harbor different self-renewing tumorigenic cell types expressing a variety of morphological and phenotypical markers, karyotypic evolution and different gene expression profiles. This suggests that the models of hepatic carcinogenesis should take into account TPC heterogeneity due to intratumour clonal evolution.

AB - Background and Aims: Increasing evidence that a number of malignancies are characterised by tumour cell heterogeneity has recently been published, but there is still a lack of data concerning liver cancers. The aim of this study was to investigate and characterise tumour-propagating cell (TPC) compartments within human hepatocellular carcinoma (HCC). Methods: After long-term culture, we identified three morphologically different tumour cell populations in a single HCC specimen, and extensively characterised them by means of flow cytometry, fluorescence microscopy, karyotyping and microarray analyses, single cell cloning, and xenotransplantation in NOD/SCID/IL2Rγ-/- mice. Results: The primary cell populations (hcc-1, -2 and -3) and two clones generated by means of limiting dilutions from hcc-1 (clone-1/7 and -1/8) differently expressed a number of tumour-associated stem cell markers, including EpCAM, CD49f, CD44, CD133, CD56, Thy-1, ALDH and CK19, and also showed different doubling times, drug resistance and tumorigenic potential. Moreover, we found that ALDH expression, in combination with CD44 or Thy-1 negativity or CD56 positivity identified subpopulations with a higher clonogenic potential within hcc-1, hcc-2 and hcc-3 primary cell populations, respectively. Karyotyping revealed the clonal evolution of the cell populations and clones within the primary tumour. Importantly, the primary tumour cell population with the greatest tumorigenic potential and drug resistance showed more chromosomal alterations than the others and contained clones with epithelial and mesenchymal features. Conclusions: Individual HCCs can harbor different self-renewing tumorigenic cell types expressing a variety of morphological and phenotypical markers, karyotypic evolution and different gene expression profiles. This suggests that the models of hepatic carcinogenesis should take into account TPC heterogeneity due to intratumour clonal evolution.

UR - http://www.scopus.com/inward/record.url?scp=79959551642&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79959551642&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0021369

DO - 10.1371/journal.pone.0021369

M3 - Article

VL - 6

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 6

M1 - e21369

ER -