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Statins affect ETS1 -overexpressing triple-negative breast cancer cells by restoring DUSP4 deficiency

View Article: PubMed Central - PubMed

ABSTRACT

We investigated the molecular mechanisms underlying statin-induced growth suppression of triple-negative breast cancer (TNBC) that overexpress the transcription factor ets proto-oncogene 1(ets-1) and downregulate dual specific protein phosphatase 4(dusp4) expression. We examined the gene expression of BC cell lines using the nCounter expression assay, MTT viability assay, cell proliferation assay and Western blot to evaluate the effects of simvastatin. Finally, we performed cell viability testing in TNBC cell line-transfected DUSP4. We demonstrated that ETS1 mRNA and protein were overexpressed in TNBC cells compared with other BC cell lines (P = <0.001) and DUSP4 mRNA was downregulated (P = <0.001). MTT viability assay showed that simvastatin had significant antitumor activity (P = 0.002 in 0.1 μM). In addition, simvastatin could restore dusp4 deficiency and suppress ets-1 expression in TNBC. Lastly, we found that si-DUSP4 RNA transfection overcame the antitumor activity of statins. MAPK pathway inhibitor, U0126 and PI3KCA inhibitor LY294002 also decreased levels of ets-1, phosphor-ERK and phosphor-AKT on Western blot assay. Accordingly, our study indicates that simvastatin potentially affects the activity of transcriptional factors such as ets-1 and dusp4 through the MAPK pathway. In conclusion, statins might be potential candidates for TNBC therapy reducing ets-1 expression via overexpression of dusp4.

No MeSH data available.


Effects of statin treatment on gene expression.(A) Quantitative real-time RT–PCR analysis of the mRNA expression levels of ETS-1 and DUSP4. (B) Western blot analysis of TNBC cell lines before and after simvastatin treatment. Blots were probed with anti-DUSP4 and anti-ETS1 antibodies.
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f3: Effects of statin treatment on gene expression.(A) Quantitative real-time RT–PCR analysis of the mRNA expression levels of ETS-1 and DUSP4. (B) Western blot analysis of TNBC cell lines before and after simvastatin treatment. Blots were probed with anti-DUSP4 and anti-ETS1 antibodies.

Mentions: In addition, we evaluated the effects of statins on dusp4 and ets-1 expression using RT-PCR and Western blot analyses (Fig. 3A,B). In HCC1806 cells, simvastatin treatment (>5 μM) resulted in decreased ets-1 mRNA expression and increased dusp4 expression (Fig. 3A). MDA-MD-231 cells were more sensitive to simvastatin than HCC1806 cells; 0.4 μM simvastatin was sufficient to affect ets-1 and dusp4 mRNA expression. Western blot analysis also revealed that the effects of simvastatin on ets-1 and dusp4 expression were dose-dependent (Fig. 3B).


Statins affect ETS1 -overexpressing triple-negative breast cancer cells by restoring DUSP4 deficiency
Effects of statin treatment on gene expression.(A) Quantitative real-time RT–PCR analysis of the mRNA expression levels of ETS-1 and DUSP4. (B) Western blot analysis of TNBC cell lines before and after simvastatin treatment. Blots were probed with anti-DUSP4 and anti-ETS1 antibodies.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5015082&req=5

f3: Effects of statin treatment on gene expression.(A) Quantitative real-time RT–PCR analysis of the mRNA expression levels of ETS-1 and DUSP4. (B) Western blot analysis of TNBC cell lines before and after simvastatin treatment. Blots were probed with anti-DUSP4 and anti-ETS1 antibodies.
Mentions: In addition, we evaluated the effects of statins on dusp4 and ets-1 expression using RT-PCR and Western blot analyses (Fig. 3A,B). In HCC1806 cells, simvastatin treatment (>5 μM) resulted in decreased ets-1 mRNA expression and increased dusp4 expression (Fig. 3A). MDA-MD-231 cells were more sensitive to simvastatin than HCC1806 cells; 0.4 μM simvastatin was sufficient to affect ets-1 and dusp4 mRNA expression. Western blot analysis also revealed that the effects of simvastatin on ets-1 and dusp4 expression were dose-dependent (Fig. 3B).

View Article: PubMed Central - PubMed

ABSTRACT

We investigated the molecular mechanisms underlying statin-induced growth suppression of triple-negative breast cancer (TNBC) that overexpress the transcription factor ets proto-oncogene 1(ets-1) and downregulate dual specific protein phosphatase 4(dusp4) expression. We examined the gene expression of BC cell lines using the nCounter expression assay, MTT viability assay, cell proliferation assay and Western blot to evaluate the effects of simvastatin. Finally, we performed cell viability testing in TNBC cell line-transfected DUSP4. We demonstrated that ETS1 mRNA and protein were overexpressed in TNBC cells compared with other BC cell lines (P = <0.001) and DUSP4 mRNA was downregulated (P = <0.001). MTT viability assay showed that simvastatin had significant antitumor activity (P = 0.002 in 0.1 μM). In addition, simvastatin could restore dusp4 deficiency and suppress ets-1 expression in TNBC. Lastly, we found that si-DUSP4 RNA transfection overcame the antitumor activity of statins. MAPK pathway inhibitor, U0126 and PI3KCA inhibitor LY294002 also decreased levels of ets-1, phosphor-ERK and phosphor-AKT on Western blot assay. Accordingly, our study indicates that simvastatin potentially affects the activity of transcriptional factors such as ets-1 and dusp4 through the MAPK pathway. In conclusion, statins might be potential candidates for TNBC therapy reducing ets-1 expression via overexpression of dusp4.

No MeSH data available.