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Transcriptional silencing of ETS-1 abrogates epithelial-mesenchymal transition resulting in reduced motility of pancreatic cancer cells.

Li C, Wang Z, Chen Y, Zhou M, Zhang H, Chen R, Shi F, Wang C, Rui Z - Oncol. Rep. (2014)

Bottom Line: In the present study, we focused on the effect of ETS-1 on epithelial-mesenchymal transition (EMT), which is characterized by reduced E-cadherin expression and increased N-cadherin expression.We detected reduced N-cadherin and vascular endothelial growth factor yet higher E-cadherin expression in the ETS-1-silenced cells compared with the control group.In addition, we observed reduced cell migration and increased adhesion in these cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu 210009, P.R. China.

ABSTRACT
v-ets erythroblastosis virus E26 oncogene homolog 1 (ETS-1) plays crucial roles in a spectrum of malignancies. ETS-1 has gained attention in cancer research for its importance in cell migration, invasion and proliferation. In the present study, we focused on the effect of ETS-1 on epithelial-mesenchymal transition (EMT), which is characterized by reduced E-cadherin expression and increased N-cadherin expression. We found that ETS-1 mRNA expression was positively correlated with N-cadherin and negatively correlated with E-cadherin mRNA expression in five pancreatic cancer cell lines. To elucidate the functionality of ETS-1 on EMT in pancreatic cancer cells, we constructed a green fluorescent protein (GFP)-expressing plasmid carrying ETS-1 short hairpin RNA (shRNA), and transfected Panc-1 cells with the plasmid. We detected reduced N-cadherin and vascular endothelial growth factor yet higher E-cadherin expression in the ETS-1-silenced cells compared with the control group. In addition, we observed reduced cell migration and increased adhesion in these cells. Our data showed that ETS-1 actively functioned as a regulator of EMT in Panc-1 cells, and provide additional evidence supporting a fundamental role for ETS-1 in metastatic pancreatic cancer cells. These results suggest that analysis of ETS-1 expression levels may provide an avenue for evaluating prognosis in pancreatic cancer.

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Related in: MedlinePlus

Efficacy of ETS-1 transcriptional silencing. Expression level of ETS-1 in the Panc-1 cells was decreased after ETS-1 shRNA transfection. (A) Brightfield and fluorescence images (magnification, ×100). (B) qRT-PCR analysis. Data are presented as mean ± SD of three independent experiments; ***P<0.001. (C) Lysates from ETS-1 shRNA- and control shNC-transfected cells were subjected to western blot analysis; GAPDH was included as a normalization control. ETS-1, v-ets erythroblastosis virus E26 oncogene homolog 1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.
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f2-or-33-02-0559: Efficacy of ETS-1 transcriptional silencing. Expression level of ETS-1 in the Panc-1 cells was decreased after ETS-1 shRNA transfection. (A) Brightfield and fluorescence images (magnification, ×100). (B) qRT-PCR analysis. Data are presented as mean ± SD of three independent experiments; ***P<0.001. (C) Lysates from ETS-1 shRNA- and control shNC-transfected cells were subjected to western blot analysis; GAPDH was included as a normalization control. ETS-1, v-ets erythroblastosis virus E26 oncogene homolog 1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Mentions: We performed ETS-1 inhibition in the Panc-1 cancer cell line, which exhibited relatively high expression of ETS-1 (Fig. 1). ETS-1 inhibition was performed using a green fluorescent protein (GFP)-expressing adenoviral vector carrying an shRNA targeting the ETS-1 gene. The success of the plasmid transfection was monitored by green fluorescence (Fig. 2A). The silencing efficiency of the ETS-1 shRNA was determined using qRT-PCR and western blot analysis. qRT-PCR showed a 70% reduction in mRNA expression in the ETS-1 shRNA-transfected cells (Fig. 2B). The level of ETS-1 protein was also significantly decreased in the ETS-1-silenced cells (Fig. 2C).


Transcriptional silencing of ETS-1 abrogates epithelial-mesenchymal transition resulting in reduced motility of pancreatic cancer cells.

Li C, Wang Z, Chen Y, Zhou M, Zhang H, Chen R, Shi F, Wang C, Rui Z - Oncol. Rep. (2014)

Efficacy of ETS-1 transcriptional silencing. Expression level of ETS-1 in the Panc-1 cells was decreased after ETS-1 shRNA transfection. (A) Brightfield and fluorescence images (magnification, ×100). (B) qRT-PCR analysis. Data are presented as mean ± SD of three independent experiments; ***P<0.001. (C) Lysates from ETS-1 shRNA- and control shNC-transfected cells were subjected to western blot analysis; GAPDH was included as a normalization control. ETS-1, v-ets erythroblastosis virus E26 oncogene homolog 1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4306275&req=5

f2-or-33-02-0559: Efficacy of ETS-1 transcriptional silencing. Expression level of ETS-1 in the Panc-1 cells was decreased after ETS-1 shRNA transfection. (A) Brightfield and fluorescence images (magnification, ×100). (B) qRT-PCR analysis. Data are presented as mean ± SD of three independent experiments; ***P<0.001. (C) Lysates from ETS-1 shRNA- and control shNC-transfected cells were subjected to western blot analysis; GAPDH was included as a normalization control. ETS-1, v-ets erythroblastosis virus E26 oncogene homolog 1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.
Mentions: We performed ETS-1 inhibition in the Panc-1 cancer cell line, which exhibited relatively high expression of ETS-1 (Fig. 1). ETS-1 inhibition was performed using a green fluorescent protein (GFP)-expressing adenoviral vector carrying an shRNA targeting the ETS-1 gene. The success of the plasmid transfection was monitored by green fluorescence (Fig. 2A). The silencing efficiency of the ETS-1 shRNA was determined using qRT-PCR and western blot analysis. qRT-PCR showed a 70% reduction in mRNA expression in the ETS-1 shRNA-transfected cells (Fig. 2B). The level of ETS-1 protein was also significantly decreased in the ETS-1-silenced cells (Fig. 2C).

Bottom Line: In the present study, we focused on the effect of ETS-1 on epithelial-mesenchymal transition (EMT), which is characterized by reduced E-cadherin expression and increased N-cadherin expression.We detected reduced N-cadherin and vascular endothelial growth factor yet higher E-cadherin expression in the ETS-1-silenced cells compared with the control group.In addition, we observed reduced cell migration and increased adhesion in these cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu 210009, P.R. China.

ABSTRACT
v-ets erythroblastosis virus E26 oncogene homolog 1 (ETS-1) plays crucial roles in a spectrum of malignancies. ETS-1 has gained attention in cancer research for its importance in cell migration, invasion and proliferation. In the present study, we focused on the effect of ETS-1 on epithelial-mesenchymal transition (EMT), which is characterized by reduced E-cadherin expression and increased N-cadherin expression. We found that ETS-1 mRNA expression was positively correlated with N-cadherin and negatively correlated with E-cadherin mRNA expression in five pancreatic cancer cell lines. To elucidate the functionality of ETS-1 on EMT in pancreatic cancer cells, we constructed a green fluorescent protein (GFP)-expressing plasmid carrying ETS-1 short hairpin RNA (shRNA), and transfected Panc-1 cells with the plasmid. We detected reduced N-cadherin and vascular endothelial growth factor yet higher E-cadherin expression in the ETS-1-silenced cells compared with the control group. In addition, we observed reduced cell migration and increased adhesion in these cells. Our data showed that ETS-1 actively functioned as a regulator of EMT in Panc-1 cells, and provide additional evidence supporting a fundamental role for ETS-1 in metastatic pancreatic cancer cells. These results suggest that analysis of ETS-1 expression levels may provide an avenue for evaluating prognosis in pancreatic cancer.

Show MeSH
Related in: MedlinePlus