The epidermal growth factor stimulates sphingosine kinase-1 expression and activity in the human mammary carcinoma cell line MCF7

Döll, Frauke; Pfeilschifter, Josef; Huwiler, Andrea

doi:10.1016/j.bbalip.2005.12.001

Cited by 88 publications

(91 citation statements)

References 59 publications

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“…To see whether the second delayed phase of SK activation depends on increased SK-1 protein expression, Western blot analyses were performed, using a specific antibody against SK-1 which was previously characterized (Döll et al 2005, Huwiler et al 2006.…”

Section: Resultsmentioning

confidence: 99%

“…Cell culturing and siRNA transfections MCF7 cells were cultured as previously reported (Döll et al 2005), and gene silencing of SKs was conducted exactly as described (Huwiler et al 2006).…”

Section: Chemicalsmentioning

confidence: 99%

“…Protein extracts were prepared exactly as previously described (Döll et al 2005). About 30 mg protein were separated by SDS-PAGE, transferred to nitrocellulose membrane, and Western blot analysis was performed as previously described using a polyclonal anti-peptide antibody against human SK-1 at a dilution of 1:2000 (Döll et al 2005).…”

Section: Western Blot Analysismentioning

confidence: 99%

“…To measure SK-1 promoter activity, a luciferase reporter gene assay was performed exactly as previously described (Huwiler et al 2006) using a Lumat LB9507 luminometer (Berthold Detection Systems, Pforzheim, Germany). SK activity assay SK-1 activity was measured exactly as previously described (Döll et al 2005, Huwiler et al 2006. In brief, 30 mg protein extracts and 10 ml of 1 mM sphingosine (dissolved in 1 mg/ml BSA) were mixed with SK-1 buffer (20 mM Tris, pH 7.4, 20% glycerol, 1 mM mercaptoethanol, 1 mM EDTA, 1 mM sodium orthovanadate, 40 mM b-glycerophosphate, 15 mM NaF, 10 mg/ml leupeptin, 10 mg/ml aprotinin, 10 mg/ ml soybean trypsin inhibitor, 1 mM phenylmethylsulfonyl fluoride) in a total volume of 190 ml.…”

Section: Promoter Studiesmentioning

confidence: 99%

“…Chemotaxis was measured in Nunc cell culture inserts (polycarbonate membrane, 10 mm, 8.0 mm pore size) exactly as described (Döll et al 2005).…”

Section: Migration Assaymentioning

confidence: 99%

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Prolactin upregulates sphingosine kinase-1 expression and activity in the human breast cancer cell line MCF7 and triggers enhanced proliferation and migration

Döll¹,

Pfeilschifter²,

Huwiler³

2007

Endocr Relat Cancer

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Sphingosine kinases (SK) catalyze the formation of sphingosine-1-phosphate (S1P) which plays a crucial role in cell growth and survival. Here, we show that prolactin (PRL) biphasically activates the SK-1, but not the SK-2 subtype, in the breast adenocarcinoma cell-line MCF7. A first peak occurs after minutes of stimulation and is followed by a second delayed activation after hours of stimulation. A similar biphasic effect on SK-1 activity is seen for 17b-estradiol (E 2 ). The delayed activation of SK-1 derives from an upregulated mRNA and protein expression and is due to increased SK-1 promoter activity and mechanistically involves STAT5 activation as well as protein kinase C and the classical mitogen-activated protein kinases. Furthermore, glucocorticoids also block both hormone-induced SK-1 expression and activity. Functionally, long-term stimulation of MCF7 cells with PRL or E 2 is well known to trigger increased cell proliferation and migration. Both hormone-induced cell responses critically involve SK-1 activation since the depletion of SK-1, but not SK-2, by siRNA transfection abolishes the hormone-induced cell proliferation and migration. In summary, our data show that PRL and E 2 cause a pronounced delayed SK-1 activation which is due to increased gene transcription, and critically determines the capability of cells to grow and move. Thus, the SK-1 may represent a novel attractive target for anti-tumor therapy.

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Section: Resultsmentioning

confidence: 99%

Section: Chemicalsmentioning

confidence: 99%

Section: Western Blot Analysismentioning

confidence: 99%

Section: Promoter Studiesmentioning

confidence: 99%

“…Chemotaxis was measured in Nunc cell culture inserts (polycarbonate membrane, 10 mm, 8.0 mm pore size) exactly as described (Döll et al 2005).…”

Section: Migration Assaymentioning

confidence: 99%

See 3 more Smart Citations

Prolactin upregulates sphingosine kinase-1 expression and activity in the human breast cancer cell line MCF7 and triggers enhanced proliferation and migration

Döll¹,

Pfeilschifter²,

Huwiler³

2007

Endocr Relat Cancer

View full text Add to dashboard Cite

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Sphingosine Kinases: Biochemistry, Regulation, and Roles

Pitman

Jarman

Leclercq

et al. 2013

Lysophospholipid Receptors

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Erratum

2009

Cancer

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It has recently been shown that docetaxel chemotherapy is effective in prolonging life in patients with prostate cancer (PCa). We have investigated potential ways of increasing the effectiveness of chemotherapy in this disease. We have previously reported that sphingosine kinase 1 (SphK1) inhibition is a key step in docetaxel‐induced apoptosis in the PC‐3 PCa cell line and that pharmacologicalSphK1 inhibition is chemosensitizing in the docetaxel‐resistant PCa LNCaP cell line. In this study we have addressed the mechanism of docetaxel‐induced apoptosis of PC‐3 cells and identified SphK1‐dependent and ‐independent components. We have shown that SphK1 inhibition by docetaxel is a two‐step process involving an initial loss of enzyme activity followed by a decrease in SphK1 gene expression. Using hormoneresistant PC‐3 and DU145 PCa cells we have demonstrated that both pharmacological and siRNA‐mediated SphK1 inhibition leads to a four‐fold decrease in the docetaxel IC50 dose. This work points out to potential ways of increasing the effectiveness of chemotherapy for PCa by SphK1 inhibition. © 2009 UICC

show abstract

The epidermal growth factor stimulates sphingosine kinase-1 expression and activity in the human mammary carcinoma cell line MCF7

Cited by 88 publications

References 59 publications

Prolactin upregulates sphingosine kinase-1 expression and activity in the human breast cancer cell line MCF7 and triggers enhanced proliferation and migration

Prolactin upregulates sphingosine kinase-1 expression and activity in the human breast cancer cell line MCF7 and triggers enhanced proliferation and migration

Sphingosine Kinases: Biochemistry, Regulation, and Roles

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