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Ataxin-1 regulates epithelial – mesenchymal transition of cervical cancer cells

View Article: PubMed Central - PubMed

ABSTRACT

The mutant form of the protein ataxin-1 (ATXN1) causes the neurodegenerative disease spinocerebellar ataxia type-1. Recently, ATXN1 was reported to enhance E-cadherin expression in the breast cancer cell line MCF-7, suggesting a potential association between ATXN1 and cancer development. In the present study, we discovered a novel mechanism through which ATXN1 regulates the epithelial–mesenchymal transition (EMT) of cancer cells. Hypoxia-induced upregulation of the Notch intracellular domain expression decreased ATXN1 expression via MDM2-associated ubiquitination and degradation. In cervical cancer cells, ATXN1 knockdown induced EMT by directly regulating Snail expression, leading to matrix metalloproteinase activation and the promotion of cell migration and invasion. These findings provide insights into a novel mechanism of tumorigenesis and will facilitate the development of new and more effective therapies for cancer.

No MeSH data available.


Related in: MedlinePlus

Inhibition of ATXN1 expression increases the migration and invasiveness of cervical cancer cell linesA. Upper panel: SiHa, CaSki, and C33A cells were transfected with siCon (sicontrol) or siATXN1 and subjected to a wound-healing assay. Lower panel: Quantification was performed by measuring the migration distances. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 100 μm. B. SiHa cells were transfected with siCon or siATXN1 and analyzed in a Matrigel invasion assay for 72 h. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 20 μm. C. The migration of HeLashATXN1-#1, HeLashATXN1-#2, and control cells was assayed in a wound-healing assay. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 100 μm. D. HeLashATXN1-#1, HeLashATXN1-#2, and control cells were analyzed in a Matrigel invasion assay for 72 h. *P<0.05, **P<0.01, ***P<0.001 vs. control group (one-way ANOVA). Scale bar: 20 μm. E. Real-time qRT–PCR analysis of MMP mRNAs in HeLashATXN1-#1, HeLashATXN1-#2, and control cells. All quantitative data are shown as the means and standard deviations of three independent experiments. *P<0.05, **P<0.01, ***P<0.001 vs. control group (one-way ANOVA). F. Proposed model for the role of ATXN1 in cervical cancer cell development. Hypoxia-induced upregulation of NICD expression induces reductions in ATXN1 and E-cadherin expression and an increase in Snail expression. Thus, ATXN1 expression decreases in the late stages of tumor development, thus inducing the EMT in cervical cancer cells.
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Figure 5: Inhibition of ATXN1 expression increases the migration and invasiveness of cervical cancer cell linesA. Upper panel: SiHa, CaSki, and C33A cells were transfected with siCon (sicontrol) or siATXN1 and subjected to a wound-healing assay. Lower panel: Quantification was performed by measuring the migration distances. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 100 μm. B. SiHa cells were transfected with siCon or siATXN1 and analyzed in a Matrigel invasion assay for 72 h. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 20 μm. C. The migration of HeLashATXN1-#1, HeLashATXN1-#2, and control cells was assayed in a wound-healing assay. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 100 μm. D. HeLashATXN1-#1, HeLashATXN1-#2, and control cells were analyzed in a Matrigel invasion assay for 72 h. *P<0.05, **P<0.01, ***P<0.001 vs. control group (one-way ANOVA). Scale bar: 20 μm. E. Real-time qRT–PCR analysis of MMP mRNAs in HeLashATXN1-#1, HeLashATXN1-#2, and control cells. All quantitative data are shown as the means and standard deviations of three independent experiments. *P<0.05, **P<0.01, ***P<0.001 vs. control group (one-way ANOVA). F. Proposed model for the role of ATXN1 in cervical cancer cell development. Hypoxia-induced upregulation of NICD expression induces reductions in ATXN1 and E-cadherin expression and an increase in Snail expression. Thus, ATXN1 expression decreases in the late stages of tumor development, thus inducing the EMT in cervical cancer cells.

Mentions: We performed a wound-healing assay to investigate the role of ATXN1 in cell migration. Transfection of the cervical cancer cell lines SiHa, CaSki, and C33A with siATXN1 promoted increased migration relative to controls (Figure 5A). We performed a Matrigel-coated transwell invasion assay to further examine the invasive properties of SiHa cells. Cells transfected with siATXN1 migrated more rapidly and were more invasive than controls (Figure 5B). Furthermore, the migration (Figure 5C) and invasion (Figure 5D) patterns of HeLashATXN1-#1 and HeLashATXN1-#2 cells were similar to those of the siATXN1 transfectants.


Ataxin-1 regulates epithelial – mesenchymal transition of cervical cancer cells
Inhibition of ATXN1 expression increases the migration and invasiveness of cervical cancer cell linesA. Upper panel: SiHa, CaSki, and C33A cells were transfected with siCon (sicontrol) or siATXN1 and subjected to a wound-healing assay. Lower panel: Quantification was performed by measuring the migration distances. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 100 μm. B. SiHa cells were transfected with siCon or siATXN1 and analyzed in a Matrigel invasion assay for 72 h. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 20 μm. C. The migration of HeLashATXN1-#1, HeLashATXN1-#2, and control cells was assayed in a wound-healing assay. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 100 μm. D. HeLashATXN1-#1, HeLashATXN1-#2, and control cells were analyzed in a Matrigel invasion assay for 72 h. *P<0.05, **P<0.01, ***P<0.001 vs. control group (one-way ANOVA). Scale bar: 20 μm. E. Real-time qRT–PCR analysis of MMP mRNAs in HeLashATXN1-#1, HeLashATXN1-#2, and control cells. All quantitative data are shown as the means and standard deviations of three independent experiments. *P<0.05, **P<0.01, ***P<0.001 vs. control group (one-way ANOVA). F. Proposed model for the role of ATXN1 in cervical cancer cell development. Hypoxia-induced upregulation of NICD expression induces reductions in ATXN1 and E-cadherin expression and an increase in Snail expression. Thus, ATXN1 expression decreases in the late stages of tumor development, thus inducing the EMT in cervical cancer cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 5: Inhibition of ATXN1 expression increases the migration and invasiveness of cervical cancer cell linesA. Upper panel: SiHa, CaSki, and C33A cells were transfected with siCon (sicontrol) or siATXN1 and subjected to a wound-healing assay. Lower panel: Quantification was performed by measuring the migration distances. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 100 μm. B. SiHa cells were transfected with siCon or siATXN1 and analyzed in a Matrigel invasion assay for 72 h. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 20 μm. C. The migration of HeLashATXN1-#1, HeLashATXN1-#2, and control cells was assayed in a wound-healing assay. *P<0.05, **P<0.01, ***P<0.001, t test. Scale bar: 100 μm. D. HeLashATXN1-#1, HeLashATXN1-#2, and control cells were analyzed in a Matrigel invasion assay for 72 h. *P<0.05, **P<0.01, ***P<0.001 vs. control group (one-way ANOVA). Scale bar: 20 μm. E. Real-time qRT–PCR analysis of MMP mRNAs in HeLashATXN1-#1, HeLashATXN1-#2, and control cells. All quantitative data are shown as the means and standard deviations of three independent experiments. *P<0.05, **P<0.01, ***P<0.001 vs. control group (one-way ANOVA). F. Proposed model for the role of ATXN1 in cervical cancer cell development. Hypoxia-induced upregulation of NICD expression induces reductions in ATXN1 and E-cadherin expression and an increase in Snail expression. Thus, ATXN1 expression decreases in the late stages of tumor development, thus inducing the EMT in cervical cancer cells.
Mentions: We performed a wound-healing assay to investigate the role of ATXN1 in cell migration. Transfection of the cervical cancer cell lines SiHa, CaSki, and C33A with siATXN1 promoted increased migration relative to controls (Figure 5A). We performed a Matrigel-coated transwell invasion assay to further examine the invasive properties of SiHa cells. Cells transfected with siATXN1 migrated more rapidly and were more invasive than controls (Figure 5B). Furthermore, the migration (Figure 5C) and invasion (Figure 5D) patterns of HeLashATXN1-#1 and HeLashATXN1-#2 cells were similar to those of the siATXN1 transfectants.

View Article: PubMed Central - PubMed

ABSTRACT

The mutant form of the protein ataxin-1 (ATXN1) causes the neurodegenerative disease spinocerebellar ataxia type-1. Recently, ATXN1 was reported to enhance E-cadherin expression in the breast cancer cell line MCF-7, suggesting a potential association between ATXN1 and cancer development. In the present study, we discovered a novel mechanism through which ATXN1 regulates the epithelial&ndash;mesenchymal transition (EMT) of cancer cells. Hypoxia-induced upregulation of the Notch intracellular domain expression decreased ATXN1 expression via MDM2-associated ubiquitination and degradation. In cervical cancer cells, ATXN1 knockdown induced EMT by directly regulating Snail expression, leading to matrix metalloproteinase activation and the promotion of cell migration and invasion. These findings provide insights into a novel mechanism of tumorigenesis and will facilitate the development of new and more effective therapies for cancer.

No MeSH data available.


Related in: MedlinePlus