Modulation of chemotactic and pro-inflammatory activities of endothelial progenitor cells by hepatocellular carcinoma

Yu Tsung Shih, Mei Cun Wang, Hsin Hsin Peng, Ting Fang Chen, Linyi Chen, Jang-Yang Chang, Jeng Jiann Chiu

Research output: Contribution to journalArticle

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Abstract

Endothelial progenitor cells (EPCs) participate in the neovascularization processes in the development of hepatocellular carcinoma (HCC). We investigated whether interactions between EPCs and HCC cells affect chemotactic and pro-inflammatory activities of EPCs. Two distinct phenotypes of circulating EPCs, i.e., myeloid-derived EPCs (colony forming unit-endothelial cells, CFU-ECs) and outgrowth EPCs (endothelial-colony forming cells, ECFCs), were co-cultured with Huh7 and Hep3B cells by using transwell chamber and IBIDI TM Culture-Inserts and μ-slide plates. Transwell and horizontal migration/invasion assays and time-lapse microscopy were used to monitor and analyze the migration and invasion of EPCs induced by these HCC cells. A human cytokine antibody array was used to compare protein expression profiles in EPCs and HCC cells. Flow cytometry and electromobility shift analysis were used to detect nuclear factor-κB (NF-κB)-DNA binding activity and pro-inflammatory adhesion molecule expression in EPCs. Ectopic full-length CC chemokine receptor 6 (CCR6) plasmid was used to transfect into ECFCs to investigate the role of CCR6 in HCC-induced EPC migration and invasion. The results show that co-culture with Huh7 and Hep3B cells induces the expression of endothelial cell (EC) markers KDR, Flt1, CD31 and VE-cadherin in CFU-ECs, but down-regulates the expressions of CD31 and VE-cadherin in ECFCs. These HCC cells induce migration and invasion of CFU-ECs, but not ECFCs, and do not affect the cell cycle distribution in these EPCs. Cytokine protein array identifies macrophage inflammatory protein-3α (MIP-3α) produced by HCC cells as a critical factor responsible for the HCC-induced chemotaxis of CFU-ECs, which highly express the specific MIP-3α counterreceptor CCR6. Overexpressing CCR6 in ECFCs significantly increases their chemotaxis in response to HCC cells. Co-culturing EPCs with HCC cells results in decreases in NF-κB binding activity and hence intracellular adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin expressions in EPCs. Our results indicate that HCC cells exert differential effects on CFU-ECs and ECFCs, with increased chemotaxis for CFU-ECs, but not ECFCs. This HCC-induced chemotaxis of CFU-ECs is mediated by MIP-3α produced by HCC cells, which targets to CCR6 on CFU-ECs. Tumors may provide a humoral microenvironment to attenuate the pro-inflammatory activity of EPCs, which might be associated with the tumor escape mechanism.

Original languageEnglish
Pages (from-to)779-793
Number of pages15
JournalCellular Signalling
Volume24
Issue number3
DOIs
Publication statusPublished - 2012 Mar 1

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Hepatocellular Carcinoma
CCR6 Receptors
Endothelial Cells
Stem Cells
Chemotaxis
Macrophage Inflammatory Proteins
Endothelial Progenitor Cells
Cell Movement
B-Form DNA
Cytokines
Tumor Escape
Protein Array Analysis
E-Selectin
Vascular Cell Adhesion Molecule-1
Coculture Techniques
Microscopy
Cell Cycle
Flow Cytometry
Plasmids
Down-Regulation

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Shih, Yu Tsung ; Wang, Mei Cun ; Peng, Hsin Hsin ; Chen, Ting Fang ; Chen, Linyi ; Chang, Jang-Yang ; Chiu, Jeng Jiann. / Modulation of chemotactic and pro-inflammatory activities of endothelial progenitor cells by hepatocellular carcinoma. In: Cellular Signalling. 2012 ; Vol. 24, No. 3. pp. 779-793.
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Modulation of chemotactic and pro-inflammatory activities of endothelial progenitor cells by hepatocellular carcinoma. / Shih, Yu Tsung; Wang, Mei Cun; Peng, Hsin Hsin; Chen, Ting Fang; Chen, Linyi; Chang, Jang-Yang; Chiu, Jeng Jiann.

In: Cellular Signalling, Vol. 24, No. 3, 01.03.2012, p. 779-793.

Research output: Contribution to journalArticle

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