Expression of Intercellular Adhesion Molecule 1 by Hepatocellular Carcinoma Stem Cells and Circulating Tumor Cells

doi:10.1053/j.gastro.2013.01.046

Gastroenterology

Volume 144, Issue 5, May 2013, Pages 1031-1041.e10

https://doi.org/10.1053/j.gastro.2013.01.046 Get rights and content

Background & Aims

Intercellular adhesion molecule 1 (ICAM-1) is believed to be involved in metastasis of hepatocellular carcinoma (HCC) cells. Cancer stem cells promote tumor relapse and metastasis. We investigated whether ICAM-1 is a marker of HCC stem cells.

Methods

Sphere formation and tumor formation assays were performed to investigate the stem cell properties of ICAM-1⁺ cells in vitro and in vivo. A specific targeting system that inhibits ICAM-1 expression and hepatitis B virus transgenic mice (M-TgHBV) were used to investigate whether inhibition of ICAM-1 reduced tumor incidence and metastasis in vivo. We used real-time polymerase chain reaction and immunoblot analysis to assess ICAM-1 and Nanog expression in tumor cell lines, and flow cytometry analysis was used to investigate ICAM-1 expression in HCC and blood samples.

Results

ICAM-1 was expressed on a minor cell population in HCC tumor cell lines, as well as in tumor tissues and circulating tumor cells isolated from patients and transgenic mice. ICAM-1⁺ tumor cells had greater sphere-forming and tumorigenic capacities and increased expression of stemness-related genes compared with ICAM-1⁻ tumor cells. The specific inhibition of ICAM-1 reduced formation and metastasis in M-TgHBV mice. ICAM-1 was found to be a marker of circulating tumor cells from patients and M-TgHBV mice. Increased numbers of CD45⁻ICAM-1⁺ cells in blood samples of patients with HCC correlated with worse clinical outcomes. The stem cell transcription factor Nanog regulated expression of ICAM-1 in HCC stem cells.

Conclusions

ICAM-1 is a marker of HCC stem cells in humans and mice; ICAM-1 inhibitors slow tumor formation and metastasis in mice. ICAM-1 expression is regulated by the stem cell transcription factor Nanog.

Section snippets

Samples

Human tumor tissues and blood samples were obtained from patients with HCC at the Eastern Hepatobiliary Surgery Hospital after obtaining informed consent. The follow-up procedures applied to these patients have been described in a previous report.²² Overall survival and disease-free survival were defined as previously described.²³ Female nude mice (4–6 weeks old) were purchased from the Transgenic Animal Research Center, Second Military Medical University. All mice were maintained in a

ICAM-1⁺ Tumor Cells Possess Characteristics of Stem/Progenitor Cells

To investigate whether ICAM-1 can be used as a CSC marker, we first determined whether an ICAM-1⁺ cell population was present in tumor cell lines. Flow cytometry analysis showed that approximately 5% of Huh7 cells and 7% of Hep3B cells expressed ICAM-1 (Figure 1A). We next examined whether the ICAM-1⁺ cells exhibited intrinsic properties of stem cells. For this purpose, ICAM-1⁺ cells were isolated from tumor cell lines, and the expression of stemness-related genes, including sox2, nanog, oct4,

Discussion

In this study, we showed that ICAM-1⁺ cells isolated from cell lines and clinical samples were capable of inducing tumors in vivo and forming spheres in vitro, indicating that the ICAM-1⁺ cells exhibited CSC properties and that ICAM-1 could be used as a potential marker for CSCs. The specific inhibition of ICAM-1 expression in vivo reduced HCC development and metastasis, which indicated that ICAM-1 could be used as a therapeutic target. Moreover, ICAM-1 was found to be transcribed directly by

Acknowledgments

The authors thank Baoyang Hu and Xiaoqing Zhang for helpful discussions and critical readings.

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: Author names in bold designate shared co-first authorship.

: Conflicts of interest The authors disclose no conflicts.

: Funding Supported by the Program for New Century Excellent Talents in University and the specially appointed Professor of Shanghai as well as by grants from the SMMU Innovation Alliance for Liver Cancer Diagnosis and Treatment (2012), the Science Fund for Creative Research Groups of China ( (81221061), China National Funds for Distinguished Young Scientists (81125018), Ministry of Technology Key Program (2008zx10002-025), National Natural Science Foundation (81071750, 30873352), and Shanghai Science and Technology Committee (10JC1417600, 10XD1405800).

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Original ResearchFull Report: Basic and Translational—LiverExpression of Intercellular Adhesion Molecule 1 by Hepatocellular Carcinoma Stem Cells and Circulating Tumor Cells

Background & Aims

Methods

Results

Conclusions

Section snippets

Samples

ICAM-1+ Tumor Cells Possess Characteristics of Stem/Progenitor Cells

Discussion

Acknowledgments

Hepatology

Pharmacol Rep

J Biol Chem