Human mesenchymal stem cell and epithelial hepatic carcinoma cell lines in admixture: concurrent stimulation of cancer-associated fibroblasts and epithelial-to-mesenchymal transition markers Journal Article

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Authors: Bhattacharya, S. D.; Mi, Z.; Talbot, L. J.; Guo, H.; Kuo, P. C.
Article Title: Human mesenchymal stem cell and epithelial hepatic carcinoma cell lines in admixture: concurrent stimulation of cancer-associated fibroblasts and epithelial-to-mesenchymal transition markers
Abstract: BACKGROUND: The microenvironments of neoplasms influence both mesenchymal stem cell differentiation into cancer-associated fibroblasts (CAF) and tumor cell line differentiation to mesenchymal phenotypes via epithelial-to-mesenchymal transition (EMT). Using direct cell-cell contact approximating the microenvironment of a neoplasm, we investigated the role of this interaction in human mesenchymal stem cells (hMSCs) and epithelial hepatic carcinoma SK-Hep1 cells by evaluating CAF differentiation and EMT. METHODS: hMSCs and SK-Hep1 cells were homogenously cultured for 12 hours with media only, OPN-R3 aptamer blockade of OPN, or RGD peptide blockade of integrin receptor, negative control mutant OPN-R3 aptamer, and RGE peptide blockade. mRNA was isolated from each subpopulation, and real-time-polymerase chain reaction was performed for CAF markers and EMT transcription factors and structural proteins. RESULTS: SK-Hep1 cells in admixture with hMSCs showed increased EMT marker vimentin expression that was ablated with OPN-R3 aptamer or RGD blockade. SK-Hep1 cells when cultured with hMSC admixture increased Snail and Slug expression that was hindered with OPN-R3 aptamer. hMSCs acquired CAF markers tenascin-c and SDF-1 in admixture that was ablated with either OPN-R3 aptamer or RGD blockade. All SK-Hep1 and hMSC negative control subpopulations were statistically equivalent to media-only groups. Fluorescence photography exhibited the critical cell-cell interfaces and acquired EMT traits of SK-Hep1. CONCLUSION: We conclude that direct interaction of cell lines closely replicates the native neoplasm microenvironment. Our experiments reveal soluble OPN or integrin receptor blockade independently prevents progression to metastatic phenotype by acquisition of CAF and EMT markers.
Keywords: Humans; Cell Line, Tumor; Surgery; Carcinoma, Hepatocellular/pathology; Coculture Techniques; Epithelial Cells/pathology; Epithelial-Mesenchymal Transition; Fibroblasts/pathology; Liver Neoplasms/pathology; Mesenchymal Stromal Cells/pathology; Microscopy, Fluorescence; Tumor Markers, Biological; Tumor Microenvironment
Journal Title: Surgery
Volume: 152
Issue: 3
ISSN: 1532-7361; 0039-6060
Publisher: Unknown  
Journal Place: United States
Date Published: 2012
Start Page: 449
End Page: 454
Language: eng
DOI/URL:

10.1016/j.surg.2012.06.011
Notes: LR: 20131114; CI: Copyright (c) 2012; GR: R01 GM065113/GM/NIGMS NIH HHS/United States; GR: R01 GM065113-A2/GM/NIGMS NIH HHS/United States; GR: T32 CA093245-09/CA/NCI NIH HHS/United States; GR: T32 GM069331/GM/NIGMS NIH HHS/United States; GR: UL1 RR024128/RR/NCRR NIH HHS/United States; JID: 0417347; 0 (Tumor Markers, Biological); NIHMS399487; OID: NLM: NIHMS399487; OID: NLM: PMC3432987; 2012/01/05 [received]; 2012/06/07 [accepted]; ppublish