Quantitative expression profile of PSGR in prostate cancer

Xu, Linda; Sun, Chen; Petrovics, György; Makarem, Mazen; Furusato, Bungo; Zhang, W; Ia, Sesterhenn; McLeod, David G.; Sun, Leon; Moul, Judd W.; Srivastava, Shiv

doi:10.1038/sj.pcan.4500836

Cited by 53 publications

(59 citation statements)

References 27 publications

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“…These data imply that a comparison of 10 000 dissected cells from sample A against 10 000 dissected cells from sample B can produce an artifactual difference of up to 1.84 C T values using cell count as the normalization strategy. Since changes in gene expression of 1.5-2 C T values (B3-to 4-fold) are considered biologically important, 30 the use of dissection cell count as a means to compare samples is limited only to those studies that do not require precise measurements. However, we did find that cell count was useful in producing 'ballpark' RNA input levels within the range that can be successfully analyzed by qRT-PCR, and thus serves as a useful first step when analyzing dissected samples.…”

Section: Resultsmentioning

confidence: 99%

Assessment of normalization strategies for quantitative RT-PCR using microdissected tissue samples

Erickson

Albert

Gillespie

et al. 2007

Laboratory Investigation

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Gene expression measurement techniques such as quantitative reverse transcriptase (qRT)-PCR require a normalization strategy to allow meaningful comparisons across biological samples. Typically, this is accomplished through the use of an endogenous housekeeping gene that is presumed to show stable expression levels in the samples under study. There is concern regarding how precisely specific genes can be measured in limited amounts of mRNA such as those from microdissected (MD) tissues. To address this issue, we evaluated three different approaches for qRT-PCR normalization of dissected samples; cell count during microdissection, total RNA measurement, and endogenous control genes. The data indicate that both cell count and total RNA are useful in calibrating input amounts at the outset of a study, but do not provide enough precision to serve as normalization standards. However, endogenous control genes can accurately determine the relative abundance of a target gene relative to the entire cellular transcriptome. Taken together, these results suggest that precise gene expression measurements can be made from MD samples if the appropriate normalization strategy is employed.

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Section: Resultsmentioning

confidence: 99%

Assessment of normalization strategies for quantitative RT-PCR using microdissected tissue samples

Erickson

Albert

Gillespie

et al. 2007

Laboratory Investigation

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“…All other primers and probes have been reported. 21,23 The TaqMan primers and probe for a-methylacyl-CoA racemase (AMACR) were 'assay on demand' (PE Applied Biosystems, Foster City, CA, USA). The expression of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was analyzed as endogenous control (Human GAPDH TaqMan mix, Applied Biosystems).…”

Section: Methodsmentioning

confidence: 99%

“…Pearson's or Spearman's correlation analyses were used to evaluate the correlation between C-MYC and other gene-expression levels (PTEN, ERG, AR, PSA/KLK3, PCA3, PMEPA1, PSGR, TMPRSS2, AMACR and LTF) in tumor tissue. 20,21,23 The prognostic significance of established clinical variables (race, pathologic stage and Gleason's score, margin status and PSA level at time of diagnosis) and C-MYC expression level were assessed by constructing univariate and multivariate Cox proportional hazards models. Disease progression-free survival curves across different C-MYC expression levels (categorized to quartile groups) were constructed with use of KaplanMeier survival method with log-rank test.…”

Section: Methodsmentioning

confidence: 99%

Overexpression of C-MYC oncogene in prostate cancer predicts biochemical recurrence

Hawksworth

Ravindranath

Chen

et al. 2010

Prostate Cancer Prostatic Dis

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145

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Alterations of chromosome 8, including amplification at 8q24 harboring the C-MYC oncogene, have been noted as one of the most common chromosomal abnormalities in prostate cancer (CaP) progression. However, the frequency of C-MYC alterations in CaP has remained uncertain. A recent study, using a new anti-MYC antibody, described prevalent upregulation of nuclear C-MYC protein expression as an early oncogenic alteration in CaP. Further, we have recently reported regulation of C-MYC expression by ERG and a significant correlation between C-MYC overexpression and TMPRSS2-ERG fusion in early stage CaP. These emerging data suggest that increased C-MYC expression may be a critical and early oncogenic event driving CaP progression. In this study, we assessed whether C-MYC mRNA overexpression in primary prostate tumors was predictive of more aggressive tumor or disease progression. Our approach was to quantitatively determine C-MYC mRNA expression levels in laser capture micro-dissected tumor cells and matched benign epithelial cells in a radical prostatectomy cohort with long follow-up data available. On the basis of our results, we conclude that elevated C-MYC expression in primary prostate tumor is biologically relevant and may be a predictor of future biochemical recurrence.

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“…PMEPA1 is a NEDD4-binding protein that was originally identified by our laboratory as a prostate-abundant, highly androgen-induced gene and was mapped to chromosome 20q13 (32,33). Human PMEPA1 exhibits amino acid homology to the mouse Nedd4-binding protein Nedd4BP (33).…”

mentioning

confidence: 99%

“…Human PMEPA1 exhibits amino acid homology to the mouse Nedd4-binding protein Nedd4BP (33). PMEPA1 protein has two PY motifs (PPPY and PPTY), which are required for binding to WW domains of NEDD4 E3 ubiquitin ligase (34).…”

mentioning

confidence: 99%

A Feedback Loop between the Androgen Receptor and a NEDD4-binding Protein, PMEPA1, in Prostate Cancer Cells

Liu

Masuda

et al. 2008

Journal of Biological Chemistry

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PMEPA1 was identified originally as a highly androgen-inducible gene with prostate-abundant expression that was restricted to prostatic epithelial cells. PMEPA1 protein is a NEDD4 (ubiquitin-protein isopeptide ligase)-binding protein, which negatively regulates prostate cancer cell growth. In this study we establish that PMEPA1 is a direct transcriptional target of the androgen receptor (AR). We also demonstrate that PMEPA1 negatively regulates AR protein levels in different cell culture models. Transient expression of PMEPA1 down-regulates AR protein levels and AR transcriptional targets in prostate cancer cells. Conversely, knockdown of PMEPA1 leads to elevated levels of AR protein, AR transcriptional targets (prostatespecific antigen), and increased cell cycle S phase. We define that the PMEPA1-dependent down-regulation of AR is because of AR ubiquitination and proteasome-mediated degradation. The mutant PMEPA1 (PY1/2 motif mutation) that is impaired in NEDD4 recruitment shows attenuated AR ubiquitination and AR protein down-regulation. These data support the hypothesis that PMEPA1 negatively regulates the stability of AR protein by enhancing AR ubiquitination and proteasome-mediated degradation through NEDD4. The effect of PMEPA1 on AR ubiquitination and degradation appears to be MDM2-independent. Thus, the PMEPA1-AR degradation pathway may represent a new androgen-dependent mechanism for regulating AR levels in prostate epithelial cells. These findings underscore that the decreased PMEPA1 expression frequently noted in prostate cancers may lead to increased AR functions and strengthen the biological role of PMEPA1 in prostate cancers.

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Quantitative expression profile of PSGR in prostate cancer

Cited by 53 publications

References 27 publications

Assessment of normalization strategies for quantitative RT-PCR using microdissected tissue samples

Assessment of normalization strategies for quantitative RT-PCR using microdissected tissue samples

Overexpression of C-MYC oncogene in prostate cancer predicts biochemical recurrence

A Feedback Loop between the Androgen Receptor and a NEDD4-binding Protein, PMEPA1, in Prostate Cancer Cells

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