Journal: PLoS ONE

Article Title: ERBB2 Increases Metastatic Potentials Specifically in Androgen-Insensitive Prostate Cancer Cells

doi: 10.1371/journal.pone.0099525

Figure Lengend Snippet: ERBB2 and RAS protein levels were assessed by Western blots using antibodies against H-RAS or ERBB2 for total cell lysates prepared from prostate cancer cells transfected with either control retroviruses ( PBP ), or retroviruses overexpressing PBP - H-RAS ( RAS ) or PBP-ERBB2 ( ERBB2 ). Blots with antibodies against actin served as loading controls. Numbers in white represent fold changes in ERBB2 or RAS protein levels in ERBB2 - or RAS -overexpressing cells relative to those in their corresponding PBP control cells after the actin normalization.

Article Snippet: Cell signaling pathways activated by the ERBB2 oncogene or the RAS oncogene are frequently found to be altered in metastatic prostate cancers.

Techniques: Western Blot, Transfection

Journal: PLoS ONE

Article Title: ERBB2 Increases Metastatic Potentials Specifically in Androgen-Insensitive Prostate Cancer Cells

doi: 10.1371/journal.pone.0099525

Figure Lengend Snippet: Cell growth rates were assessed by cell counting every 12 hours or 24 hours for various prostate cancer cells that were transfected with control retroviruses ( PBP ), or retroviruses overexpressing either PBP-H-RAS ( RAS ) or PBP-ERBB2 ( ERBB2 ).

Article Snippet: Cell signaling pathways activated by the ERBB2 oncogene or the RAS oncogene are frequently found to be altered in metastatic prostate cancers.

Techniques: Cell Counting, Transfection

Journal: PLoS ONE

Article Title: ERBB2 Increases Metastatic Potentials Specifically in Androgen-Insensitive Prostate Cancer Cells

doi: 10.1371/journal.pone.0099525

Figure Lengend Snippet: Cell migration rates were estimated by a wound healing assay for prostate cancer cells that were transfected with either control retroviruses ( PBP ), or retroviruses overexpressing PBP - H-RAS ( RAS ) or PBP-ERBB2 ( ERBB2 ). Left panels showed percentages of wounds remained at different time points. The percentages of wounds were estimated based on the average of 12 measurements on each plate reflecting measurements of four evenly distributed sections on each of the three wounds/scratches on each plate. Data were presented as means ± SD from three replicates. Right panels showed representative images taken at different time points. All images were taken at the same scale with a scale bar of 200 µM displayed in the first image.

Article Snippet: Cell signaling pathways activated by the ERBB2 oncogene or the RAS oncogene are frequently found to be altered in metastatic prostate cancers.

Techniques: Migration, Wound Healing Assay, Transfection

Journal: PLoS ONE

Article Title: ERBB2 Increases Metastatic Potentials Specifically in Androgen-Insensitive Prostate Cancer Cells

doi: 10.1371/journal.pone.0099525

Figure Lengend Snippet: Cell motilities were assessed by a transwell-based motility assay for prostate cancer cells that were transfected with either control retroviruses ( PBP ), or retroviruses overexpressing PBP - H-RAS ( RAS ) or PBP-ERBB2 ( ERBB2 ). Each bar graph showed the relative numbers of cells that have passed through the transwell inserts/membranes, which were stained and counted under a microscope for at least 10 fields per insert. Data were presented as means ± SD from three replicates. Representative images taken at a given time point were shown underneath each bar graph. All images were taken at the same scale with a scale bar of 200 µM displayed in the first image.

Article Snippet: Cell signaling pathways activated by the ERBB2 oncogene or the RAS oncogene are frequently found to be altered in metastatic prostate cancers.

Techniques: Motility Assay, Transfection, Staining, Microscopy

Journal: PLoS ONE

Article Title: ERBB2 Increases Metastatic Potentials Specifically in Androgen-Insensitive Prostate Cancer Cells

doi: 10.1371/journal.pone.0099525

Figure Lengend Snippet: Cell invasiveness was assessed by a transwell-based invasion assay for prostate cancer cells that were transfected with either control retroviruses ( PBP ), or retroviruses overexpressing PBP - H-RAS ( RAS ) or PBP-ERBB2 ( ERBB2 ). Each bar graph showed the numbers of cells that have passed through the collagen matrix either 72 hours (for PC3 cells) or 96 hours (for DU145 cells) after plating. Transwell inserts were stained and invading cells were counted for the entire inserts. Data were presented as means ± SD from three replicates. Representative images were shown underneath each bar graph. All images were taken at the same scale with a scale bar of 200 µM displayed in the first image.

Article Snippet: Cell signaling pathways activated by the ERBB2 oncogene or the RAS oncogene are frequently found to be altered in metastatic prostate cancers.

Techniques: Transwell Invasion Assay, Transfection, Staining

Journal: PLoS ONE

Article Title: ERBB2 Increases Metastatic Potentials Specifically in Androgen-Insensitive Prostate Cancer Cells

doi: 10.1371/journal.pone.0099525

Figure Lengend Snippet: (A) Senescence-associated β-galactosidase activities were assessed by X-gal staining for human prostate cancer cell lines transfected with either control retroviruses ( PBP ) or retroviruses overexpressing PBP-H-RAS ( RAS ). PC3 cells expressing an extremely high level of RAS ( Hi-RAS ) were included for comparisons. Senescent BJ human skin fibroblast cells were used as a positive control for X-gal staining. Representative images were shown with representative X-gal-positive cells being marked with red arrows. Inserts in each image showed magnified, representative X-gal-positive cells. ( B ) Quantifications of data collected from panel ( A ). Data were presented as means ± SD from three replicates. ( C ) RAS protein levels assessed by Western blot analysis with antibodies against H-RAS for parental PC3 cells (P), PC3 cells transfected with control retroviruses ( PBP ), or PC3 cells transfected with the PBP-H-RAS retroviruses overexpressing a moderate level of RAS ( RAS ) or a much higher level of RAS ( Hi-RAS ). A blot using antibodies against actin was used as a loading control. Numbers in white represent RAS protein levels in fold changes in ERBB2 - or RAS -overexpressing cells relative to those in PBP control cells after the actin normalization. Scare bar: 100 µM.

Article Snippet: Cell signaling pathways activated by the ERBB2 oncogene or the RAS oncogene are frequently found to be altered in metastatic prostate cancers.

Techniques: Staining, Transfection, Expressing, Positive Control, Western Blot

Journal: PLoS ONE

Article Title: ERBB2 Increases Metastatic Potentials Specifically in Androgen-Insensitive Prostate Cancer Cells

doi: 10.1371/journal.pone.0099525

Figure Lengend Snippet: Protein levels for various kinases and their phosphorylated forms were assessed by Western blot analyses for parental prostate cancer cells that were transfected either with control retroviruses ( PBP ), or with retroviruses overexpressing PBP-H-RAS ( RAS ) or PBP-ERBB2 ( ERBB2 ). Actin was used as a loading control. Numbers in white represent protein levels in fold changes relative to those in PBP control cells after the actin normalization.

Article Snippet: Cell signaling pathways activated by the ERBB2 oncogene or the RAS oncogene are frequently found to be altered in metastatic prostate cancers.

Techniques: Western Blot, Transfection