Journal: PLoS ONE

Article Title: The Response of the Prostate to Circulating Cholesterol: Activating Transcription Factor 3 (ATF3) as a Prominent Node in a Cholesterol-Sensing Network

doi: 10.1371/journal.pone.0039448

Figure Lengend Snippet: ( A ) A provisional network was generated from integration of two microarray data sets. Node color represents increases (red), no significant changes (yellow), and decreases (green) in gene expression in murine prostate tissue after cholesterol alteration as ascertained by cDNA microarray. Changes in RNA expression levels of the corresponding nodes in LNCaP cells are shown as colored node boundaries (donut shape) and the color represents increases (red), no significant change (yellow), and decreases (green) in gene expression under CDM conditions compared to control. Arrows indicate direct activation, T-shaped lines direct repression, dashed arrows indirect activation, and lines physical interaction. ( B ) Gene expression under Normo and Hyper conditions ( in vivo ). To verify in vivo microarray data obtained from SCID experiments, mRNA levels of the indicated genes were determined. GAPDH expression was used to normalize gene expression. Error bars represent SD (n = 3). ( C ) Gene expression under Control and Cholesterol-depleted conditions ( in vitro ). LNCaP cells were incubated in CDM for 0, 3 or 16 h, and mRNA levels of the indicated genes were measured by RT-PCR analysis to validate cDNA microarray data. Error bars represent SD (n = 3). * p <0.05 (Student’s t-test).

Article Snippet: LNCaP human prostate tumor cells were purchased from American Type Culture Collection (ATCC, Manassas, VA) and maintained in RPMI1640 media (Invitrogen, Carlsbad, CA) supplemented with 10% FBS and 1% Penicillin/Streptomycin at 37°C under 5% CO 2 .

Techniques: Generated, Microarray, Gene Expression, RNA Expression, Control, Activation Assay, In Vivo, Expressing, In Vitro, Incubation, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: The Response of the Prostate to Circulating Cholesterol: Activating Transcription Factor 3 (ATF3) as a Prominent Node in a Cholesterol-Sensing Network

doi: 10.1371/journal.pone.0039448

Figure Lengend Snippet: ( A ) RT-PCR analysis in vivo . ATF3 levels are reduced in all prostatic lobes from Hyper mice, compared to those from the Normo group (AP = anterior prostate; VP = ventral prostate; DLP = dorsal prostate). ( B ) Immunoblot analysis. Immunoblot of PrEC lysates showed induction of ATF3 protein by CDM (left panel) and by β-cyclodextrin (right panel). MG132, a proteasome inhibitor, also increased ATF3 expression. ( C ) Immunofluorescence analysis. Induction of ATF3 protein by CDM in LNCaP cells as shown by IF. LNCaP cells were treated with CDM for 18 h, stained with anti-ATF3 antibody and nuclei were counterstained with DAPI (left panel: ATF3; middle panel: DAPI; right panel: overlay). ( D ) RT-PCR analysis. ATF3 mRNA levels in LNCaP cells treated with CDM were normalized to levels of GAPDH. RT-PCR analysis shows induction of ATF3 mRNA levels by CDM. ( E–F ) Promoter reporter analysis. A full-length ATF3 promoter was cloned into a luciferase reporter vector and transfected into LNCaP (D) or PrEC (E). Cells were then incubated in Control and CDM medium. ATF3 promoter activity was plotted as arbitrary units (± SD) after normalization with total protein concentration.

Article Snippet: LNCaP human prostate tumor cells were purchased from American Type Culture Collection (ATCC, Manassas, VA) and maintained in RPMI1640 media (Invitrogen, Carlsbad, CA) supplemented with 10% FBS and 1% Penicillin/Streptomycin at 37°C under 5% CO 2 .

Techniques: Reverse Transcription Polymerase Chain Reaction, In Vivo, Western Blot, Expressing, Immunofluorescence, Staining, Clone Assay, Luciferase, Plasmid Preparation, Transfection, Incubation, Control, Activity Assay, Protein Concentration

Journal: Genomics Data

Article Title: Expression-profiling of apoptosis induced by ablation of the long ncRNA TRPM2-AS in prostate cancer cell

doi: 10.1016/j.gdata.2014.10.020

Figure Lengend Snippet: Validation of microarray data by qRT-PCR. Ten transcripts were quantified by qRT-PCR both in control and in TRPM2-AS KO PC3 cells. The resulting expression fold change is plotted against the expression fold change obtained from the Illumina HumanHT-12 V3.0 microarray data. A correlation coefficient of 0.97 was found between the two datasets.

Article Snippet: The human, androgen-independent prostate cancer cell line PC3 was obtained from ATCC (CRL-1435, ATCC).

Techniques: Biomarker Discovery, Microarray, Quantitative RT-PCR, Control, Expressing

Journal: Genomics Data

Article Title: Expression-profiling of apoptosis induced by ablation of the long ncRNA TRPM2-AS in prostate cancer cell

doi: 10.1016/j.gdata.2014.10.020

Figure Lengend Snippet:

Article Snippet: The human, androgen-independent prostate cancer cell line PC3 was obtained from ATCC (CRL-1435, ATCC).

Techniques: Expressing, Microarray, Gene Expression

Journal: The FASEB Journal

Article Title: 12-HETER1/GPR31, a high-affinity 12( S )-hydroxyeicosatetraenoic acid receptor, is significantly up-regulated in prostate cancer and plays a critical role in prostate cancer progression

doi: 10.1096/fj.201500076

Figure Lengend Snippet: 12-HETER1 expression positively correlates with aggressiveness and progression of prostate tumors. A) Expression of 12-HETER1 in prostate carcinoma cells. Immunocytochemistry on HEK293 cells expressing 12-HETER1. a, b) Staining with 12-HETER1 antibody was optimized on HEK293 cells transfected with 12-HETER1 (a) or pcDNA (b) vector. Strong brown staining was detected in HEK293/12-HETER1 cells (a) compared to negative control (b). c, d) Immunohistochemistry to detect 12-HETER1 in 2 cases of human prostate cancer tissue. c) Benign gland (N, left corner, arrow) with rare cells showing positive staining, and neoplastic glands (T, GS-3/4, arrowhead) showing intense brown staining (arrowhead); original magnification, ×10. d) Strong staining in area (upper left, arrow) with mixed GS-3/4 tumor area, weak staining in area (lower right, arrowhead) with GS-3 tumor; original magnification, ×10. Images are representatives from 7 samples. B) Array analysis of 12-HETER1 expression. a, b) Immunohistochemistry to assay 12-HETER1 expression on 2 human prostate tissue microarrays. Expression of 12-HETER1 is significantly up-regulated in disease stage II/III human prostate tumor specimens (n = 8, P = 0.007, Student’s t test. c, d) Representative images of normal (c) and prostate tumor (d) specimens.

Article Snippet: Next we used qPCR to determine 12-HETER1 expression in 2 commercially available human prostate tissue microarrays from OriGene Technologies, HPRT101 and HPRT102 ( ).

Techniques: Expressing, Immunocytochemistry, Staining, Transfection, Plasmid Preparation, Negative Control, Immunohistochemistry