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MicroRNA-155 expression is independently predictive of outcome in chordoma.

Osaka E, Kelly AD, Spentzos D, Choy E, Yang X, Shen JK, Yang P, Mankin HJ, Hornicek FJ, Duan Z - Oncotarget (2015)

Bottom Line: Regulatory activity of miR-155 was assessed using bioinformatic tools. miR-155 expression levels were validated by reverse transcription-polymerase chain reaction.Inhibition of miR-155 expression suppressed proliferation, and the migratory and invasive activities of chordoma cells.These results collectively indicate that miR-155 expression may serve not only as a prognostic marker, but also as a potential therapeutic target in chordoma.

View Article: PubMed Central - PubMed

Affiliation: Sarcoma Biology Laboratory, Department of Orthopaedic Surgery, Massachusetts General Hospital, Boston, MA 02114, USA.

ABSTRACT

Background: Chordoma pathogenesis remains poorly understood. In this study, we aimed to evaluate the relationships between microRNA-155 (miR-155) expression and the clinicopathological features of chordoma patients, and to evaluate the functional role of miR-155 in chordoma.

Methods: The miRNA expression profiles were analyzed using miRNA microarray assays. Regulatory activity of miR-155 was assessed using bioinformatic tools. miR-155 expression levels were validated by reverse transcription-polymerase chain reaction. The relationships between miR-155 expression and the clinicopathological features of chordoma patients were analyzed. Proliferative, migratory and invasive activities were assessed by MTT, wound healing, and Matrigel invasion assays, respectively.

Results: The miRNA microarray assay revealed miR-155 to be highly expressed and biologically active in chordoma. miR-155 expression in chordoma tissues was significantly elevated, and this expression correlated significantly with disease stage (p = 0.036) and the presence of metastasis (p = 0.035). miR-155 expression also correlated significantly with poor outcomes for chordoma patients (hazard ratio, 5.32; p = 0.045). Inhibition of miR-155 expression suppressed proliferation, and the migratory and invasive activities of chordoma cells.

Conclusions: We have shown miR-155 expression to independently affect prognosis in chordoma. These results collectively indicate that miR-155 expression may serve not only as a prognostic marker, but also as a potential therapeutic target in chordoma.

No MeSH data available.


Related in: MedlinePlus

Inhibition of miR-155 decreases proliferation, migration, and invasion of chordoma cells(A, B) Cell proliferation assay results for U-CH1 cells transfected with miR-155 inhibitor at concentrations between 0 nM and 80 nM, and incubated for 24–96 h. (C, D) miR-155 antagonism resulted in the inhibition of the migratory activity of U-CH1 chordoma cells after transfection with either miR-155 or non-specific miR inhibitor at 40 nM. (C) micrographs of chordoma U-CH1 cells at 0 h, 8 h and 24 h after wounding. (D) The coverage rate of U-CH1 cells for each time point and condition in the wound healing assay. (E, F) Invasive activity assay results for U-CH1 cells transfected with miR-155 inhibitor. (E) Micrographs of chordoma U-CH1 cells transfected with 40 nM miR-155 inhibitor. (F) Average numbers of invasive chordoma cells among those transfected with 10–80 nM miR-155. *p < 0.01.
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Figure 5: Inhibition of miR-155 decreases proliferation, migration, and invasion of chordoma cells(A, B) Cell proliferation assay results for U-CH1 cells transfected with miR-155 inhibitor at concentrations between 0 nM and 80 nM, and incubated for 24–96 h. (C, D) miR-155 antagonism resulted in the inhibition of the migratory activity of U-CH1 chordoma cells after transfection with either miR-155 or non-specific miR inhibitor at 40 nM. (C) micrographs of chordoma U-CH1 cells at 0 h, 8 h and 24 h after wounding. (D) The coverage rate of U-CH1 cells for each time point and condition in the wound healing assay. (E, F) Invasive activity assay results for U-CH1 cells transfected with miR-155 inhibitor. (E) Micrographs of chordoma U-CH1 cells transfected with 40 nM miR-155 inhibitor. (F) Average numbers of invasive chordoma cells among those transfected with 10–80 nM miR-155. *p < 0.01.

Mentions: To examine whether down-regulation of miR-155 inhibits the growth of chordoma cells, we conducted Methyl thiazolyl tetrazorium (MTT) assays to evaluate altered in vitro proliferation. Chordoma cells were transfected with the miR-155 inhibitor at various concentrations (10 nM, 20 nM, 40 nM, 60 nM, and 80 nM) for 24–96 h. The MTT assays revealed that miR-155 inhibitor transfected chordoma cells showed time-dependent inhibition of cellular growth at the concentrations tested, whereas non-specific miR inhibitor transfected cells and control cells continued to grow for the duration of the observation period (Fig. 5A). After transfection with miR-155 inhibitor or non-specific miR inhibitor at a different concentration and incubation for 72 h, the miR-155 inhibitor transfected cells showed significant and dose-dependent inhibition of cellular growth (Fig. 5B). These results demonstrated that down-regulation of miR-155 inhibits cell proliferation both dose- and time-dependently in chordoma cell lines in vitro.


MicroRNA-155 expression is independently predictive of outcome in chordoma.

Osaka E, Kelly AD, Spentzos D, Choy E, Yang X, Shen JK, Yang P, Mankin HJ, Hornicek FJ, Duan Z - Oncotarget (2015)

Inhibition of miR-155 decreases proliferation, migration, and invasion of chordoma cells(A, B) Cell proliferation assay results for U-CH1 cells transfected with miR-155 inhibitor at concentrations between 0 nM and 80 nM, and incubated for 24–96 h. (C, D) miR-155 antagonism resulted in the inhibition of the migratory activity of U-CH1 chordoma cells after transfection with either miR-155 or non-specific miR inhibitor at 40 nM. (C) micrographs of chordoma U-CH1 cells at 0 h, 8 h and 24 h after wounding. (D) The coverage rate of U-CH1 cells for each time point and condition in the wound healing assay. (E, F) Invasive activity assay results for U-CH1 cells transfected with miR-155 inhibitor. (E) Micrographs of chordoma U-CH1 cells transfected with 40 nM miR-155 inhibitor. (F) Average numbers of invasive chordoma cells among those transfected with 10–80 nM miR-155. *p < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 5: Inhibition of miR-155 decreases proliferation, migration, and invasion of chordoma cells(A, B) Cell proliferation assay results for U-CH1 cells transfected with miR-155 inhibitor at concentrations between 0 nM and 80 nM, and incubated for 24–96 h. (C, D) miR-155 antagonism resulted in the inhibition of the migratory activity of U-CH1 chordoma cells after transfection with either miR-155 or non-specific miR inhibitor at 40 nM. (C) micrographs of chordoma U-CH1 cells at 0 h, 8 h and 24 h after wounding. (D) The coverage rate of U-CH1 cells for each time point and condition in the wound healing assay. (E, F) Invasive activity assay results for U-CH1 cells transfected with miR-155 inhibitor. (E) Micrographs of chordoma U-CH1 cells transfected with 40 nM miR-155 inhibitor. (F) Average numbers of invasive chordoma cells among those transfected with 10–80 nM miR-155. *p < 0.01.
Mentions: To examine whether down-regulation of miR-155 inhibits the growth of chordoma cells, we conducted Methyl thiazolyl tetrazorium (MTT) assays to evaluate altered in vitro proliferation. Chordoma cells were transfected with the miR-155 inhibitor at various concentrations (10 nM, 20 nM, 40 nM, 60 nM, and 80 nM) for 24–96 h. The MTT assays revealed that miR-155 inhibitor transfected chordoma cells showed time-dependent inhibition of cellular growth at the concentrations tested, whereas non-specific miR inhibitor transfected cells and control cells continued to grow for the duration of the observation period (Fig. 5A). After transfection with miR-155 inhibitor or non-specific miR inhibitor at a different concentration and incubation for 72 h, the miR-155 inhibitor transfected cells showed significant and dose-dependent inhibition of cellular growth (Fig. 5B). These results demonstrated that down-regulation of miR-155 inhibits cell proliferation both dose- and time-dependently in chordoma cell lines in vitro.

Bottom Line: Regulatory activity of miR-155 was assessed using bioinformatic tools. miR-155 expression levels were validated by reverse transcription-polymerase chain reaction.Inhibition of miR-155 expression suppressed proliferation, and the migratory and invasive activities of chordoma cells.These results collectively indicate that miR-155 expression may serve not only as a prognostic marker, but also as a potential therapeutic target in chordoma.

View Article: PubMed Central - PubMed

Affiliation: Sarcoma Biology Laboratory, Department of Orthopaedic Surgery, Massachusetts General Hospital, Boston, MA 02114, USA.

ABSTRACT

Background: Chordoma pathogenesis remains poorly understood. In this study, we aimed to evaluate the relationships between microRNA-155 (miR-155) expression and the clinicopathological features of chordoma patients, and to evaluate the functional role of miR-155 in chordoma.

Methods: The miRNA expression profiles were analyzed using miRNA microarray assays. Regulatory activity of miR-155 was assessed using bioinformatic tools. miR-155 expression levels were validated by reverse transcription-polymerase chain reaction. The relationships between miR-155 expression and the clinicopathological features of chordoma patients were analyzed. Proliferative, migratory and invasive activities were assessed by MTT, wound healing, and Matrigel invasion assays, respectively.

Results: The miRNA microarray assay revealed miR-155 to be highly expressed and biologically active in chordoma. miR-155 expression in chordoma tissues was significantly elevated, and this expression correlated significantly with disease stage (p = 0.036) and the presence of metastasis (p = 0.035). miR-155 expression also correlated significantly with poor outcomes for chordoma patients (hazard ratio, 5.32; p = 0.045). Inhibition of miR-155 expression suppressed proliferation, and the migratory and invasive activities of chordoma cells.

Conclusions: We have shown miR-155 expression to independently affect prognosis in chordoma. These results collectively indicate that miR-155 expression may serve not only as a prognostic marker, but also as a potential therapeutic target in chordoma.

No MeSH data available.


Related in: MedlinePlus