MiR-675 mediates downregulation of twist1 and rb in afp-secreting hepatocellular carcinoma

J. M. Hernandez, A. Elahi, C. W. Clark, J. Wang, L. A. Humphries, B. Centeno, G. Bloom, B. C. Fuchs, T. Yeatman, David Shibata

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Background. Alpha-fetoprotein (AFP)-secreting hepatocellular carcinomas (HCC) represent a genetically distinct subset of tumors often associated with a worse prognosis. However, the molecular mechanisms that underlie these phenotypic differences remain poorly understood. Methods. HCC tumor samples from 27 patients were profiled using the Affymetrix 133 Plus 2.0 GeneChips. GeneGO Metacore software was used to identify altered biologic pathways. Expression validation was confirmed by RT-PCR. Manipulation of miR-675 by overexpression and antagomir-mediated knockdown was carried out with subsequent evaluation of effects on cell behavior by cell cycle, proliferation, invasion, and growth in soft agar assays. Results. We identified a strong relationship between primary tumor H19 gene expression and elevated serum AFP. H19 has recently been identified to encode microRNA-675 (miR-675), and we confirmed the relationship in an independent sample of patients. Pathway analyses of the effect of miR-675 overexpression in hepatoma cells revealed a predominant upregulation of cell adhesion and cell cycle initiation pathways. We have demonstrated that miR-675 mediates increases in proliferation and an accumulation of cells with tetraploid DNA content associated with a repression of Rb. We also demonstrated that overexpression of miR-675 alters cellular morphology, reduces invasive potential, and increases anchorage-independent growth capacity. These findings are consistent with a mesenchymal-to-epithelial transition, associated with a reduction in the expression of the key EMT mediator, Twist1. Conclusions. Expression of the miR-675 in hepatocellular carcinoma links a dramatic upregulation of proliferative and growth capacity with inhibition of motility in HCC cells.

Original languageEnglish (US)
JournalAnnals of Surgical Oncology
Volume20
Issue number3 SUPPL.
DOIs
StatePublished - Jul 18 2013

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MicroRNAs
Hepatocellular Carcinoma
Down-Regulation
alpha-Fetoproteins
Cell Cycle
Up-Regulation
Growth
Cell Proliferation
Neoplasms
Epithelial-Mesenchymal Transition
Tetraploidy
Cell Adhesion
Agar
Software
Gene Expression
Polymerase Chain Reaction
DNA
Serum

All Science Journal Classification (ASJC) codes

  • Surgery
  • Oncology

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MiR-675 mediates downregulation of twist1 and rb in afp-secreting hepatocellular carcinoma. / Hernandez, J. M.; Elahi, A.; Clark, C. W.; Wang, J.; Humphries, L. A.; Centeno, B.; Bloom, G.; Fuchs, B. C.; Yeatman, T.; Shibata, David.

In: Annals of Surgical Oncology, Vol. 20, No. 3 SUPPL., 18.07.2013.

Research output: Contribution to journalArticle

Hernandez, JM, Elahi, A, Clark, CW, Wang, J, Humphries, LA, Centeno, B, Bloom, G, Fuchs, BC, Yeatman, T & Shibata, D 2013, 'MiR-675 mediates downregulation of twist1 and rb in afp-secreting hepatocellular carcinoma', Annals of Surgical Oncology, vol. 20, no. 3 SUPPL.. https://doi.org/10.1245/s10434-013-3106-3
Hernandez, J. M. ; Elahi, A. ; Clark, C. W. ; Wang, J. ; Humphries, L. A. ; Centeno, B. ; Bloom, G. ; Fuchs, B. C. ; Yeatman, T. ; Shibata, David. / MiR-675 mediates downregulation of twist1 and rb in afp-secreting hepatocellular carcinoma. In: Annals of Surgical Oncology. 2013 ; Vol. 20, No. 3 SUPPL.
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abstract = "Background. Alpha-fetoprotein (AFP)-secreting hepatocellular carcinomas (HCC) represent a genetically distinct subset of tumors often associated with a worse prognosis. However, the molecular mechanisms that underlie these phenotypic differences remain poorly understood. Methods. HCC tumor samples from 27 patients were profiled using the Affymetrix 133 Plus 2.0 GeneChips. GeneGO Metacore software was used to identify altered biologic pathways. Expression validation was confirmed by RT-PCR. Manipulation of miR-675 by overexpression and antagomir-mediated knockdown was carried out with subsequent evaluation of effects on cell behavior by cell cycle, proliferation, invasion, and growth in soft agar assays. Results. We identified a strong relationship between primary tumor H19 gene expression and elevated serum AFP. H19 has recently been identified to encode microRNA-675 (miR-675), and we confirmed the relationship in an independent sample of patients. Pathway analyses of the effect of miR-675 overexpression in hepatoma cells revealed a predominant upregulation of cell adhesion and cell cycle initiation pathways. We have demonstrated that miR-675 mediates increases in proliferation and an accumulation of cells with tetraploid DNA content associated with a repression of Rb. We also demonstrated that overexpression of miR-675 alters cellular morphology, reduces invasive potential, and increases anchorage-independent growth capacity. These findings are consistent with a mesenchymal-to-epithelial transition, associated with a reduction in the expression of the key EMT mediator, Twist1. Conclusions. Expression of the miR-675 in hepatocellular carcinoma links a dramatic upregulation of proliferative and growth capacity with inhibition of motility in HCC cells.",
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AU - Hernandez, J. M.

AU - Elahi, A.

AU - Clark, C. W.

AU - Wang, J.

AU - Humphries, L. A.

AU - Centeno, B.

AU - Bloom, G.

AU - Fuchs, B. C.

AU - Yeatman, T.

AU - Shibata, David

PY - 2013/7/18

Y1 - 2013/7/18

N2 - Background. Alpha-fetoprotein (AFP)-secreting hepatocellular carcinomas (HCC) represent a genetically distinct subset of tumors often associated with a worse prognosis. However, the molecular mechanisms that underlie these phenotypic differences remain poorly understood. Methods. HCC tumor samples from 27 patients were profiled using the Affymetrix 133 Plus 2.0 GeneChips. GeneGO Metacore software was used to identify altered biologic pathways. Expression validation was confirmed by RT-PCR. Manipulation of miR-675 by overexpression and antagomir-mediated knockdown was carried out with subsequent evaluation of effects on cell behavior by cell cycle, proliferation, invasion, and growth in soft agar assays. Results. We identified a strong relationship between primary tumor H19 gene expression and elevated serum AFP. H19 has recently been identified to encode microRNA-675 (miR-675), and we confirmed the relationship in an independent sample of patients. Pathway analyses of the effect of miR-675 overexpression in hepatoma cells revealed a predominant upregulation of cell adhesion and cell cycle initiation pathways. We have demonstrated that miR-675 mediates increases in proliferation and an accumulation of cells with tetraploid DNA content associated with a repression of Rb. We also demonstrated that overexpression of miR-675 alters cellular morphology, reduces invasive potential, and increases anchorage-independent growth capacity. These findings are consistent with a mesenchymal-to-epithelial transition, associated with a reduction in the expression of the key EMT mediator, Twist1. Conclusions. Expression of the miR-675 in hepatocellular carcinoma links a dramatic upregulation of proliferative and growth capacity with inhibition of motility in HCC cells.

AB - Background. Alpha-fetoprotein (AFP)-secreting hepatocellular carcinomas (HCC) represent a genetically distinct subset of tumors often associated with a worse prognosis. However, the molecular mechanisms that underlie these phenotypic differences remain poorly understood. Methods. HCC tumor samples from 27 patients were profiled using the Affymetrix 133 Plus 2.0 GeneChips. GeneGO Metacore software was used to identify altered biologic pathways. Expression validation was confirmed by RT-PCR. Manipulation of miR-675 by overexpression and antagomir-mediated knockdown was carried out with subsequent evaluation of effects on cell behavior by cell cycle, proliferation, invasion, and growth in soft agar assays. Results. We identified a strong relationship between primary tumor H19 gene expression and elevated serum AFP. H19 has recently been identified to encode microRNA-675 (miR-675), and we confirmed the relationship in an independent sample of patients. Pathway analyses of the effect of miR-675 overexpression in hepatoma cells revealed a predominant upregulation of cell adhesion and cell cycle initiation pathways. We have demonstrated that miR-675 mediates increases in proliferation and an accumulation of cells with tetraploid DNA content associated with a repression of Rb. We also demonstrated that overexpression of miR-675 alters cellular morphology, reduces invasive potential, and increases anchorage-independent growth capacity. These findings are consistent with a mesenchymal-to-epithelial transition, associated with a reduction in the expression of the key EMT mediator, Twist1. Conclusions. Expression of the miR-675 in hepatocellular carcinoma links a dramatic upregulation of proliferative and growth capacity with inhibition of motility in HCC cells.

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