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Open Access Research article

Antitumor activity of phenethyl isothiocyanate in HER2-positive breast cancer models

Parul Gupta and Sanjay K Srivastava*

Author Affiliations

Department of Biomedical Sciences and Cancer Biology Center, Texas Tech University Health Sciences Center, Amarillo, TX, USA

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BMC Medicine 2012, 10:80  doi:10.1186/1741-7015-10-80

Published: 24 July 2012

Additional files

Additional file 1:

Figure S1. Phenethyl isothiocyanate (PEITC) induces histone associated fragmentation in breast cancer cells. Apoptosis induction was measured by enzyme-linked immunosorbent assay (ELISA) cell death detection method in (A) MDA-MB-231 and (B) MCF-7 (n = 3). Each experiment was repeated more than three times independently. *Statistically different when compared with control (P < 0.05).

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Additional file 2:

Figure S2. Change in HER2 expression modulates the effect of phenethyl isothiocyanate (PEITC). (A) Effect of PEITC in HER2-silenced MDA-MB-231 cells. At 48 h after transfection of cells with HER2 siRNA, cells were treated with or without 10 μM PEITC for 24 h. Apoptosis was measured by enzyme-linked immunosorbent assay (ELISA) cell death detection method after silencing HER2 in MDA-MB-231 (n = 15). (B) Effect of HER2 overexpression on apoptosis induction in MCF-7 by PEITC treatment. After 48 h of HER2 transfection, cells were treated with or without 10 μM PEITC for 24 h. The means of three independent experiments performed in triplicate are shown. The induction of apoptosis by ELISA cell death detection method in HER2 overexpressing MCF-7 cells (n = 3).

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Additional file 3:

Figure S3. Change in HER2 expression modulates the effect of phenethyl isothiocyanate (PEITC). (A) Comparative effect of PEITC treatment on MDA-MB-231 cells with stable overexpression of HER2 relative to parent cells. Cells were treated for 24 h with 10 μM PEITC and apoptosis measured by enzyme-linked immunosorbent assay (ELISA) cell death detection method in MDA-MB-231 parent cells and the cells with stable overexpression of HER2, after treatment with 10 μM PEITC for 24 h. (B) Comparative effect of PEITC treatment in MCF-7 cells with stable overexpression of HER2 relative to the parent cells. Cells were treated for 24 h with 10 μM PEITC and apoptosis measured by ELISA cell death detection method in MCF-7 parent cells and the cells with stable overexpression of HER2, after treatment with PEITC (10 μM) for 24 h. The figures are representative of at least three independent experiments with eight replicates. *Statistically different compared with control (P < 0.05).

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Additional file 4:

Figure S4. Phenethyl isothiocyanate (PEITC) suppresses the growth of MDA-MB-231 (high HER2 (HH)) tumors by inhibiting HER2 in SCID/NOD mice. About 5 × 106 MDA-MB-231 (HH) cells were subcutaneously implanted into the right flanks of SCID/NOD mice. Once each mouse had a tumor of about 150 mm3, mice started receiving 12 μmol of PEITC by oral gavage every day. About 20 micron sections were obtained from snap frozen tumor tissues for tumor analysis. Immunofluorescence for HER2, phosphorylated signal transducer and activator of transcription 3 (p-STAT3) (Y-705) and cleaved caspase 3 in tumor sections from control and PEITC treated mice. The red staining represents the expression of HER2, p-STAT3 (Y705) and cleaved caspase 3.

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