Epithelial-to-mesenchymal transition of murine liver tumor cells promotes invasion

Wei Ding, Hanning You, Hien Dang, Francis LeBlanc, Vivian Galicia, Shelly C. Lu, Bangyan Stiles, C. Bart Rountree

Research output: Contribution to journalArticle

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Abstract

Epithelial-to-mesenchymal transition (EMT) is predicted to play a critical role in metastatic disease in hepatocellular carcinoma. In this study, we used a novel murine model of EMT to elucidate a mechanism of tumor progression and metastasis. A total of 2 3 106 liver cells isolated from Pten loxp/loxp/Alb-Cre+ mice, expanded from a single. CD133+CD45- cell clone, passage 0 (P0), were sequentially transplanted to obtain two passages of tumor cells, P1 and P2. Cells were analyzed for gene expression using micro-array and real-time polymerase chain reaction. Functional analysis included cell proliferation, migration, and invasion in vitro and orthotopic tumor metastasis assays in vivo. Although P0, P1, and P2 each formed tumors consistent with mixed liver epithelium, within the P2 cells, two distinct cell types were clearly visible: cells with epithelial morphology similar to P0 cells and cells with fibroblastoid morphology. These P2 mesenchymal cells demonstrated increased locomotion on wound healing; increased cell invasion on Matrigel basement membrane; increased EMT-associated gene expression of Snail1, Zeb1, and Zeb2; and down-regulated E-cadherin. P2 mesenchymal cells demonstrated significantly faster tumor growth in vivo compared with P2 epithelial counter-parts, with invasion of intestine, pancreas, spleen, and lymph nodes. Furthermore, P2 mesenchymal cells secreted high levels of hepatocyte growth factor (HGF), which we propose acts in a paracrine fashion to drive epithelial cells to undergo EMT. In addition, a second murine liver cancer stem cell line with methionine adenosyltransferase 1a deficiency acquired EMT after sequential transplantations, indicating that EMT was not restricted to Pten-deleted tumors. Conclusion: EMT is associated with a high rate of liver tumor proliferation, invasion, and metastasis in vivo, which is driven by HGF secreted from mesenchymal tumor cells in a feed-forward mechanism. (HEPATOLOGY 2010;52:945-953)

Original languageEnglish (US)
Pages (from-to)945-953
Number of pages9
JournalHepatology
Volume52
Issue number3
DOIs
StatePublished - Sep 1 2010

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Epithelial-Mesenchymal Transition
Liver
Neoplasms
Hepatocyte Growth Factor
Neoplasm Metastasis
Epithelial Cells
Gene Expression
Neoplastic Stem Cells
Cadherins
Locomotion
Liver Neoplasms
Basement Membrane
Wound Healing
Intestines
Cell Movement
Real-Time Polymerase Chain Reaction
Pancreas
Hepatocellular Carcinoma
Spleen
Epithelium

All Science Journal Classification (ASJC) codes

  • Hepatology

Cite this

Ding, W., You, H., Dang, H., LeBlanc, F., Galicia, V., Lu, S. C., ... Rountree, C. B. (2010). Epithelial-to-mesenchymal transition of murine liver tumor cells promotes invasion. Hepatology, 52(3), 945-953. https://doi.org/10.1002/hep.23748
Ding, Wei ; You, Hanning ; Dang, Hien ; LeBlanc, Francis ; Galicia, Vivian ; Lu, Shelly C. ; Stiles, Bangyan ; Rountree, C. Bart. / Epithelial-to-mesenchymal transition of murine liver tumor cells promotes invasion. In: Hepatology. 2010 ; Vol. 52, No. 3. pp. 945-953.
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abstract = "Epithelial-to-mesenchymal transition (EMT) is predicted to play a critical role in metastatic disease in hepatocellular carcinoma. In this study, we used a novel murine model of EMT to elucidate a mechanism of tumor progression and metastasis. A total of 2 3 106 liver cells isolated from Pten loxp/loxp/Alb-Cre+ mice, expanded from a single. CD133+CD45- cell clone, passage 0 (P0), were sequentially transplanted to obtain two passages of tumor cells, P1 and P2. Cells were analyzed for gene expression using micro-array and real-time polymerase chain reaction. Functional analysis included cell proliferation, migration, and invasion in vitro and orthotopic tumor metastasis assays in vivo. Although P0, P1, and P2 each formed tumors consistent with mixed liver epithelium, within the P2 cells, two distinct cell types were clearly visible: cells with epithelial morphology similar to P0 cells and cells with fibroblastoid morphology. These P2 mesenchymal cells demonstrated increased locomotion on wound healing; increased cell invasion on Matrigel basement membrane; increased EMT-associated gene expression of Snail1, Zeb1, and Zeb2; and down-regulated E-cadherin. P2 mesenchymal cells demonstrated significantly faster tumor growth in vivo compared with P2 epithelial counter-parts, with invasion of intestine, pancreas, spleen, and lymph nodes. Furthermore, P2 mesenchymal cells secreted high levels of hepatocyte growth factor (HGF), which we propose acts in a paracrine fashion to drive epithelial cells to undergo EMT. In addition, a second murine liver cancer stem cell line with methionine adenosyltransferase 1a deficiency acquired EMT after sequential transplantations, indicating that EMT was not restricted to Pten-deleted tumors. Conclusion: EMT is associated with a high rate of liver tumor proliferation, invasion, and metastasis in vivo, which is driven by HGF secreted from mesenchymal tumor cells in a feed-forward mechanism. (HEPATOLOGY 2010;52:945-953)",
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Ding, W, You, H, Dang, H, LeBlanc, F, Galicia, V, Lu, SC, Stiles, B & Rountree, CB 2010, 'Epithelial-to-mesenchymal transition of murine liver tumor cells promotes invasion', Hepatology, vol. 52, no. 3, pp. 945-953. https://doi.org/10.1002/hep.23748

Epithelial-to-mesenchymal transition of murine liver tumor cells promotes invasion. / Ding, Wei; You, Hanning; Dang, Hien; LeBlanc, Francis; Galicia, Vivian; Lu, Shelly C.; Stiles, Bangyan; Rountree, C. Bart.

In: Hepatology, Vol. 52, No. 3, 01.09.2010, p. 945-953.

Research output: Contribution to journalArticle

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AU - Ding, Wei

AU - You, Hanning

AU - Dang, Hien

AU - LeBlanc, Francis

AU - Galicia, Vivian

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AU - Stiles, Bangyan

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AB - Epithelial-to-mesenchymal transition (EMT) is predicted to play a critical role in metastatic disease in hepatocellular carcinoma. In this study, we used a novel murine model of EMT to elucidate a mechanism of tumor progression and metastasis. A total of 2 3 106 liver cells isolated from Pten loxp/loxp/Alb-Cre+ mice, expanded from a single. CD133+CD45- cell clone, passage 0 (P0), were sequentially transplanted to obtain two passages of tumor cells, P1 and P2. Cells were analyzed for gene expression using micro-array and real-time polymerase chain reaction. Functional analysis included cell proliferation, migration, and invasion in vitro and orthotopic tumor metastasis assays in vivo. Although P0, P1, and P2 each formed tumors consistent with mixed liver epithelium, within the P2 cells, two distinct cell types were clearly visible: cells with epithelial morphology similar to P0 cells and cells with fibroblastoid morphology. These P2 mesenchymal cells demonstrated increased locomotion on wound healing; increased cell invasion on Matrigel basement membrane; increased EMT-associated gene expression of Snail1, Zeb1, and Zeb2; and down-regulated E-cadherin. P2 mesenchymal cells demonstrated significantly faster tumor growth in vivo compared with P2 epithelial counter-parts, with invasion of intestine, pancreas, spleen, and lymph nodes. Furthermore, P2 mesenchymal cells secreted high levels of hepatocyte growth factor (HGF), which we propose acts in a paracrine fashion to drive epithelial cells to undergo EMT. In addition, a second murine liver cancer stem cell line with methionine adenosyltransferase 1a deficiency acquired EMT after sequential transplantations, indicating that EMT was not restricted to Pten-deleted tumors. Conclusion: EMT is associated with a high rate of liver tumor proliferation, invasion, and metastasis in vivo, which is driven by HGF secreted from mesenchymal tumor cells in a feed-forward mechanism. (HEPATOLOGY 2010;52:945-953)

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