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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.


Apoptosis assay.MDA-MB-231 (A) and HCC1806 (B) cells were transfected with control siRNA or DUSP4-targeting siRNA. At 24 h post-transfection, simvastatin was added and cells were incubated an additional 48 h. The distribution of annexin V-FITC/PI staining is shown. Data are expressed as averages for each time point and were calculated using the results from three independent experiments.
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f5: Apoptosis assay.MDA-MB-231 (A) and HCC1806 (B) cells were transfected with control siRNA or DUSP4-targeting siRNA. At 24 h post-transfection, simvastatin was added and cells were incubated an additional 48 h. The distribution of annexin V-FITC/PI staining is shown. Data are expressed as averages for each time point and were calculated using the results from three independent experiments.

Mentions: To investigate the mechanism of simvastatin-induced apoptosis, we transfected HCC1806 and MDA-MB-231 cells with DUSP4-targeting siRNA and analyzed apoptosis by FACS (Fig. 5). As expected, simvastatin induced apoptosis in cells transfected with control siRNA. In contrast, cells transfected with DUSP4 siRNA exhibited decreased simvastatin-induced apoptosis. In MDA-MB-231 cells, DUSP4 siRNA transfection inhibited apoptosis in the presence of 0.5, 1, and 5 μM simvastatin (P < 0.05), whereas DUSP4 siRNA transfection suppressed apoptosis in HCC1806 cells induced by 1 and 5 μM simvastatin (P < 0.05).


Statins affect ETS1 -overexpressing triple-negative breast cancer cells by restoring DUSP4 deficiency
Apoptosis assay.MDA-MB-231 (A) and HCC1806 (B) cells were transfected with control siRNA or DUSP4-targeting siRNA. At 24 h post-transfection, simvastatin was added and cells were incubated an additional 48 h. The distribution of annexin V-FITC/PI staining is shown. Data are expressed as averages for each time point and were calculated using the results from three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f5: Apoptosis assay.MDA-MB-231 (A) and HCC1806 (B) cells were transfected with control siRNA or DUSP4-targeting siRNA. At 24 h post-transfection, simvastatin was added and cells were incubated an additional 48 h. The distribution of annexin V-FITC/PI staining is shown. Data are expressed as averages for each time point and were calculated using the results from three independent experiments.
Mentions: To investigate the mechanism of simvastatin-induced apoptosis, we transfected HCC1806 and MDA-MB-231 cells with DUSP4-targeting siRNA and analyzed apoptosis by FACS (Fig. 5). As expected, simvastatin induced apoptosis in cells transfected with control siRNA. In contrast, cells transfected with DUSP4 siRNA exhibited decreased simvastatin-induced apoptosis. In MDA-MB-231 cells, DUSP4 siRNA transfection inhibited apoptosis in the presence of 0.5, 1, and 5 μM simvastatin (P < 0.05), whereas DUSP4 siRNA transfection suppressed apoptosis in HCC1806 cells induced by 1 and 5 μM simvastatin (P < 0.05).

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&thinsp;=&thinsp;&lt;0.001) and DUSP4 mRNA was downregulated (P&thinsp;=&thinsp;&lt;0.001). MTT viability assay showed that simvastatin had significant antitumor activity (P&thinsp;=&thinsp;0.002 in 0.1&thinsp;&mu;M). 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.