Characterization of CD133+ hepatocellular carcinoma cells as cancer stem/progenitor cells

doi:10.1016/j.bbrc.2006.10.128

Biochemical and Biophysical Research Communications

Volume 351, Issue 4, 29 December 2006, Pages 820-824

https://doi.org/10.1016/j.bbrc.2006.10.128 Get rights and content

Abstract

The CD133 antigen, identified as a hematopoietic stem cell marker, appears in various human embryonic epithelia including the neural tube, gut, and kidney. We herein investigated whether CD133⁺ cells isolated from human hepatocellular carcinoma cell lines possess cancer stem/progenitor cell-like properties. Among the three cell lines studied, the CD133 antigen was found to be expressed only on the surface of Huh-7 cells. CD133⁺ cells from Huh-7 performed a higher in vitro proliferative potential and lower mRNA expressions of mature hepatocyte markers, glutamine synthetase and cytochrome P450 3A4, than CD133⁻ population of Huh-7 cells. When either CD133⁺ or CD133⁻ cells were subcutaneously injected into SCID mice, CD133⁺ cells formed tumors, whereas CD133⁻ cells induced either a very small number of tumors or none at all. Taken together, the identification of CD133⁺ cells could thus be a potentially powerful tool to investigate the tumorigenic process in the hepatoma system and to also develop effective therapies targeted against hepatocellular carcinoma.

Section snippets

Materials and methods

Cell lines and culture conditions. Huh-7 (a human hepatoma cell line) and HepG2 cells (a human hepatoblastoma cell line) obtained from the Japanese Collection of Research Bioresources (Tokyo, Japan) were cultured in RPMI 1640 medium (Sigma–Aldrich, Irvine, UK) containing 1% penicillin and streptomycin (Gibco-BRL, Rockville, MD, USA), 12.5 mM Hepes buffer (Sigma–Aldrich), and supplemented with 0.2% lactoalbumin (Sigma–Aldrich) or 10% heat-inactivated fetal bovine serum (Sigma–Aldrich),

FACS analysis for the cell surface markers on human HCC cell lines

Huh-7, HepG2, and Hc cell lines were stained with fluorescence-conjugated primary antibodies against surface markers for hematopoietic stem cell (CD34, CD133), steel factor receptor (CD29, CD44), or stem cell factor receptor c-Kit (CD117), and were subjected to flow cytometry. HepG2 and Hc did not contain either CD34⁺ or CD133⁺ cells, but CD133⁺ cells were detected in 46.7% of Huh-7 cells. (Fig. 1) Only Hc cells were stained for CD44. Treatment with growth factors such as hepatocyte growth

Discussion

Stem cells are functionally defined as self-renewing and multipotent cells that exhibit multilineage differentiation [18], [19]. Somatic stem/progenitor cells are thought to self-renew in order to generate all of the mature cell types of a particular tissue through differentiation. However, the rigorous identification and isolation of tissue-specific stem/progenitor cells has so far been accomplished in only a few organ systems [9], [20], because a prospective such study on somatic stem cells

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Citation Excerpt :
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Conclusion: Incorporating intratumoral OV6 expression and CD68⁺ TAMs infiltration with established clinical indicators may serve as a promising prognostic signature for HCC, and could more accurately predict the clinical outcomes for HCC patients after liver resection.

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^☆: Abbreviations: CSCs, cancer stem cells; HCC, hepatocellular carcinoma; SCID, severe combined immunodeficient; FITC, fluorescein isothiocyanate; PE, phycoerythrin; APC, allophycocyanin; Alb, albumin; AFP, alfa-fetoprotein; GS, glutamine synthetase; CYP3A4, cytochrome P450 3A4.

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Characterization of CD133+ hepatocellular carcinoma cells as cancer stem/progenitor cells☆

Abstract

Section snippets

Materials and methods

FACS analysis for the cell surface markers on human HCC cell lines

Discussion

Cell

Immunity

Blood

J. Biol. Chem.

Heterogeneity in breast cancer and the problem of relevance of findings

Anal. Cell. Pathol.

Genome-wide views of cancer

N. Engl. J. Med.

Tumor heterogeneity

Cancer Res.

Primary bioassay of human tumor stem cells

Science

Growth characteristics of a mouse plasma cell tumor