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Transcriptomic and Functional Pathway Analysis of Human Cervical Carcinoma Cancer Cells Response to Microtubule Inhibitor.

Wang J, Yan B, Liu SM, Sun H, Pan Y, Guan D, Zhang X, Xu J, Ma H - J Cancer (2015)

Bottom Line: We selected the concentration of vinblastine inducing 30% cell death for analyses assessing the DNA content, gene expression and transcriptional gene regulation of VBL-treated KB-3 cells.Transcriptomic and hierarchical clustering analysis demonstrated that treatment of KB-3 cells with VBL altered the expression of a diverse group of genes with G2/M arrest, which regulated by four oncogenic or tumor suppresser transcription factors (AP1, NFKB1, RELA, and TP53).Northern blots also confirmed that KRT-7, FN14, IER3, and ID1 were deregulated in VBL-treated KB-3 cells.

View Article: PubMed Central - PubMed

Affiliation: 1. Scientific Research Center, Shanghai Public Health Clinical Center, 2901 Caolang Road, Jinshan District, Shanghai 201508, China ; 2. Department of Translational Molecular Pathology, The University of Texas, M.D. Anderson Cancer Center, Houston, TX 77030, USA.

ABSTRACT

Background: There clearly is a need for effective chemotherapy for early-stage, high-risk patients with human cervical carcinoma. Vinblastine (VBL) is a key microtubule inhibitor, but unproven in its mechanisms as an important antitumor agent in cervical carcinoma.

Methods: We selected the concentration of vinblastine inducing 30% cell death for analyses assessing the DNA content, gene expression and transcriptional gene regulation of VBL-treated KB-3 cells.

Results: Transcriptomic and hierarchical clustering analysis demonstrated that treatment of KB-3 cells with VBL altered the expression of a diverse group of genes with G2/M arrest, which regulated by four oncogenic or tumor suppresser transcription factors (AP1, NFKB1, RELA, and TP53). Functional pathway analysis revealed the disease response to the biological effects of vinblastine in cervical carcinoma chemotherapy including protein ubiquitination pathway, RhoGDI signaling, integrin signaling, agranulocyte adhesion and biapedesis, and actin nucleation pathways. Northern blots also confirmed that KRT-7, FN14, IER3, and ID1 were deregulated in VBL-treated KB-3 cells.

Conclusion: Transcriptional time series profiles and a functional pathway analysis of VBL-treated KB-3 cells will provide a new strategy for improving microtubule inhibitor chemotherapy for cervical carcinoma.

No MeSH data available.


Related in: MedlinePlus

Effect of vinblastine (VBL) on cellular DNA content. KB-3 cells untreated and treated with 2.5 x 10-4 µM of vinblastine for 2, 4, 8, 12, 24, and 48 h. Cells were subjected to DNA content analysis by flow cytometry with PI staining. A-D: KB-3 cells treated with 2.5 x 10-4 µM of VBL for 0, 2, 12 and 24h. E: Data analysis of Effect of VBL on cellular DNA content in KB-3 cells treated with 2.5 x 10-4 µM of VBL for 0, 2, 4, 8, 12, 24, and 48 h.
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Figure 1: Effect of vinblastine (VBL) on cellular DNA content. KB-3 cells untreated and treated with 2.5 x 10-4 µM of vinblastine for 2, 4, 8, 12, 24, and 48 h. Cells were subjected to DNA content analysis by flow cytometry with PI staining. A-D: KB-3 cells treated with 2.5 x 10-4 µM of VBL for 0, 2, 12 and 24h. E: Data analysis of Effect of VBL on cellular DNA content in KB-3 cells treated with 2.5 x 10-4 µM of VBL for 0, 2, 4, 8, 12, 24, and 48 h.

Mentions: First, we tested the cytotoxicity of vinblastine in human cervical carcinoma KB-3 cells using Sulforhodamine B (SRB) assay as described previously 8. After KB-3 cells were treated with VBL for 48 h, the median lethal dose (LD50) and the concentration of vinblastine at which 30% cell death was reached (LD30) in KB-3 cells were 6.8 x 10-4 µM and 2.5 x 10-4 µM, respectively. The flow cytometry assay provides information regarding cell cycle phase sensitivity to apoptosis is based on bivariate analysis of their DNA content 9. Next, the concentration of LD30 was selected for analyses assessing the DNA content of vinblastine-treated KB-3 cells, which was examined via propidium iodide (PI) staining and flow cytometry. At 0, 2, 4, 8, 12, 24, and 48 h post-treatment with 2.5 x 10-4 µM of vinblastine, KB-3 cells arrested predominately at G2/M phase of the cell cycle and the number of cells in the G1 phase had diminished (Fig. 1). With a longer duration of treatment (24 h), a much greater percentage of KB-3 cells at G2/M phase. These results indicate that KB-3 arrested at G2/M in response to vinblastine treatment.


Transcriptomic and Functional Pathway Analysis of Human Cervical Carcinoma Cancer Cells Response to Microtubule Inhibitor.

Wang J, Yan B, Liu SM, Sun H, Pan Y, Guan D, Zhang X, Xu J, Ma H - J Cancer (2015)

Effect of vinblastine (VBL) on cellular DNA content. KB-3 cells untreated and treated with 2.5 x 10-4 µM of vinblastine for 2, 4, 8, 12, 24, and 48 h. Cells were subjected to DNA content analysis by flow cytometry with PI staining. A-D: KB-3 cells treated with 2.5 x 10-4 µM of VBL for 0, 2, 12 and 24h. E: Data analysis of Effect of VBL on cellular DNA content in KB-3 cells treated with 2.5 x 10-4 µM of VBL for 0, 2, 4, 8, 12, 24, and 48 h.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4543753&req=5

Figure 1: Effect of vinblastine (VBL) on cellular DNA content. KB-3 cells untreated and treated with 2.5 x 10-4 µM of vinblastine for 2, 4, 8, 12, 24, and 48 h. Cells were subjected to DNA content analysis by flow cytometry with PI staining. A-D: KB-3 cells treated with 2.5 x 10-4 µM of VBL for 0, 2, 12 and 24h. E: Data analysis of Effect of VBL on cellular DNA content in KB-3 cells treated with 2.5 x 10-4 µM of VBL for 0, 2, 4, 8, 12, 24, and 48 h.
Mentions: First, we tested the cytotoxicity of vinblastine in human cervical carcinoma KB-3 cells using Sulforhodamine B (SRB) assay as described previously 8. After KB-3 cells were treated with VBL for 48 h, the median lethal dose (LD50) and the concentration of vinblastine at which 30% cell death was reached (LD30) in KB-3 cells were 6.8 x 10-4 µM and 2.5 x 10-4 µM, respectively. The flow cytometry assay provides information regarding cell cycle phase sensitivity to apoptosis is based on bivariate analysis of their DNA content 9. Next, the concentration of LD30 was selected for analyses assessing the DNA content of vinblastine-treated KB-3 cells, which was examined via propidium iodide (PI) staining and flow cytometry. At 0, 2, 4, 8, 12, 24, and 48 h post-treatment with 2.5 x 10-4 µM of vinblastine, KB-3 cells arrested predominately at G2/M phase of the cell cycle and the number of cells in the G1 phase had diminished (Fig. 1). With a longer duration of treatment (24 h), a much greater percentage of KB-3 cells at G2/M phase. These results indicate that KB-3 arrested at G2/M in response to vinblastine treatment.

Bottom Line: We selected the concentration of vinblastine inducing 30% cell death for analyses assessing the DNA content, gene expression and transcriptional gene regulation of VBL-treated KB-3 cells.Transcriptomic and hierarchical clustering analysis demonstrated that treatment of KB-3 cells with VBL altered the expression of a diverse group of genes with G2/M arrest, which regulated by four oncogenic or tumor suppresser transcription factors (AP1, NFKB1, RELA, and TP53).Northern blots also confirmed that KRT-7, FN14, IER3, and ID1 were deregulated in VBL-treated KB-3 cells.

View Article: PubMed Central - PubMed

Affiliation: 1. Scientific Research Center, Shanghai Public Health Clinical Center, 2901 Caolang Road, Jinshan District, Shanghai 201508, China ; 2. Department of Translational Molecular Pathology, The University of Texas, M.D. Anderson Cancer Center, Houston, TX 77030, USA.

ABSTRACT

Background: There clearly is a need for effective chemotherapy for early-stage, high-risk patients with human cervical carcinoma. Vinblastine (VBL) is a key microtubule inhibitor, but unproven in its mechanisms as an important antitumor agent in cervical carcinoma.

Methods: We selected the concentration of vinblastine inducing 30% cell death for analyses assessing the DNA content, gene expression and transcriptional gene regulation of VBL-treated KB-3 cells.

Results: Transcriptomic and hierarchical clustering analysis demonstrated that treatment of KB-3 cells with VBL altered the expression of a diverse group of genes with G2/M arrest, which regulated by four oncogenic or tumor suppresser transcription factors (AP1, NFKB1, RELA, and TP53). Functional pathway analysis revealed the disease response to the biological effects of vinblastine in cervical carcinoma chemotherapy including protein ubiquitination pathway, RhoGDI signaling, integrin signaling, agranulocyte adhesion and biapedesis, and actin nucleation pathways. Northern blots also confirmed that KRT-7, FN14, IER3, and ID1 were deregulated in VBL-treated KB-3 cells.

Conclusion: Transcriptional time series profiles and a functional pathway analysis of VBL-treated KB-3 cells will provide a new strategy for improving microtubule inhibitor chemotherapy for cervical carcinoma.

No MeSH data available.


Related in: MedlinePlus