Results: 4

1.
Fig. 3

Fig. 3. Correlation between IGF2 LOI and IGF2 expression. From: Insulin-like Growth Factor-2 (IGF2) Loss of Imprinting Marks a Field Defect Within Human Prostates Containing Cancer.

Gene expression was analyzed using qPCR and FLuPE utilized for imprinting analyses. Correlation analyses were performed using Spearman’s correlation. (A) Tumor samples (r = −0.49, P = 0.09), (B) distant TA tissues (r = 0.58, P = 0.017).

Sachin Bhusari, et al. Prostate. 2011 November;71(15):10.1002/pros.21379.
2.
Fig. 2

Fig. 2. IGF2 and H19 expression in tumor and TA tissues. From: Insulin-like Growth Factor-2 (IGF2) Loss of Imprinting Marks a Field Defect Within Human Prostates Containing Cancer.

Gene expression was measured using quantitative PCR(qPCR) in adjacent and distant TA tissues relative to the tumor for 18 microdissected sample sets (A), IGF2 mRNA expression (B), H19 mRNA expression (C). In contrast to predicted results, a positive correlation between IGF2 and H19 expression in distant TA tissues was seen (r = 0.82, P = 0.0001). No correlation was found between IGF2 and H19 gene expressionin tumors (data not shown) (* = P < 0.05).

Sachin Bhusari, et al. Prostate. 2011 November;71(15):10.1002/pros.21379.
3.
Fig. 1

Fig. 1. From: Insulin-like Growth Factor-2 (IGF2) Loss of Imprinting Marks a Field Defect Within Human Prostates Containing Cancer.

(A) Schematic showing prostate tumor, adjacent and distant normal tissue microdissection. Radical prostatectomy samples containing tumor foci were sectioned and samples microdissected for subsequent analyses. Adjacent and distant tumor-associated (TA) tissues did not contain histologic cancer. Samples were assessed in a three-dimensional pattern. (B) Fluorescent primer extension (FluPE) assay for IGF2 loss ofimprinting (LOI). Analysis of 9 informative sample sets using FluPE was performed and image analysis done using ImageJ software. LOI in prostate tumor foci ranged from 32 to 68% (mean 45%). Mean LOI in adjacent and distant TA regions was 39 and 38%, respectively. Analyses were performed in duplicate.

Sachin Bhusari, et al. Prostate. 2011 November;71(15):10.1002/pros.21379.
4.
Fig. 4

Fig. 4. Analysis of methylation status in paired prostate tumors and TA tissues. From: Insulin-like Growth Factor-2 (IGF2) Loss of Imprinting Marks a Field Defect Within Human Prostates Containing Cancer.

(A) Alterations in IGF2 expression have been linked to changes in methylation within DMR0 of the IGF2 promoter, as well as within CTCF#6 in the imprint control region (ICR) [24,54]. Methylation was analyzed using pyrosequencing specific primers. Tissues analyzed included prostate tumors with low or high IGF2 expression and distant TA tissues. An additional set of tissues from bladder, kidney, and prostates without cancer (NTA) that demonstrated maintained IGF2 imprinting were also analyzed for methylation. (B) At DMR0, tumors with low IGF2 expression demonstrate significantly decreased methylation compared to distant TA tissues. (C) CpG methylation analysis of CTCF6 in NTA tissues. TA prostate tissues from men with associated cancer (n = 6) demonstrating LOI is compared to imprinted NTA tissues (n = 7). A trend towards increased methylation in distant TA tissues is noted with significance seen at CpG 10 (P = 0.02).

Sachin Bhusari, et al. Prostate. 2011 November;71(15):10.1002/pros.21379.

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