Clinical trial NCT02460861: Intraoperational Prostate Loge Biopsies (iPROLOGX) After Radical Prostatovesiculectomy
Official title: Intraoperational Prostate Loge Biopsies (iPROLOGX) After Radical Prostatovesiculectomy (RPVE) in Prostate Cancer (PCA) Patients for Molecular Tumor Marker Analysis
Study type: INTERVENTIONAL
Overall status: UNKNOWN
Phase: NA
Conditions: Prostate Cancer
Keywords: Prostate Cancer
Molecular staging
GSTP1
Surgical margins
prostatovesiculectomy
Lead sponsor: University of Magdeburg
Sponsor class: OTHER
Collaborators: name: St. Antonius Hospital Gronau
First posted: 2015-06-03
Enrollment: 279
Sex: MALE
Standard ages: CHILD
ADULT
OLDER_ADULT
Accepts healthy volunteers: no
Allocation: NON_RANDOMIZED
Intervention model: PARALLEL
Primary purpose: BASIC_SCIENCE

Brief summary: This project is about the detection of occult tumor cells in surgical margins of radical prostatovesiculectomy by analysing the methylation status of Glutathione S-transferase P 1 (GSTP1). After gland excision specimens are obtained from 9 defined areas of the prostatic fossa. The biopsies are divided into two parts. One part used for histopathological analysis and the other part for moleculargenetic analysis. Results will be correlated e.g. with tumor stage, Gleason Score and prostate specific antigen (PSA).

The prostate-cancer-negative control group with bladder cancer.

Detailed description: This project is about the detection of occult tumor cells in surgical margins of radical prostatovesiculectomy by analysing the methylation status of Glutathione S-transferase P 1 (GSTP1). After gland excision specimens are obtained from 9 defined areas of the prostatic fossa. The biopsies are divided into two parts. One part used for histopathological analysis and the other part for moleculargenetic analysis. Results will be correlated e.g. with tumor stage, Gleason Score and prostate specific antigen (PSA).

The prostate-cancer-negative control group with bladder cancer.

DNA ISOLATION

DNA from biopsies stored by -80°C was isolated by using innuPREP DNA mini Kit (Analytik Jena, Jena, Germany) following protocol 1 of the manufacturer's instructions. DNA was eluted with 50 µl elution buffer. Concentration and purity were analysed by using Nanodrop 2000.

DNA BISULFITE MODIFICATION

DNA was modified by using EpiTect Bisulfite Kit (QIAGEN, Hilden, Germany) according to manufacturer's instructions. Samples were eluted once with 20 µl elution buffer.

QUANTITATIVE METHYLATION SPECIFIC PCR

Methylation status of GSTP1 is analysed by quantitative methylation-specific PCR (Q-MSP) using StepOnePlus Real-Time PCR System and StepOne Software v2.1 from Applied Biosystems (Darmstadt, Germany). Q-MSP was performed in duplicate analysing genes Actin and GSTP1. The primers' and testing probes' sequences used to amplify and detect hypermethylated GSTP1 were: 5'-AgTTgCgCggCgATTTC (forward primer), 5'-gCCCCAATACTAAATCACgACg (reverse primer) and 5'-CggTCgACgTTCggggTgTAgCg (taqman probe), labelled with fluorescence dye FAM. The primers' and testing probes' sequences used to amplify and detect Actin were: 5'-TggTgATggAggAggTTTAgTAAgT (forward primer), 5'-AACCAATAAAACCTACTCCTCCCTTAA (reverse primer),5'-ACCACCACCCAACACACAATAACAAACACA (taqman probe), labelled with fluorescence dye VIC.

The Q-MSP was carried out at 50°C for 2 min., 95°C for 15 min. followed by 50 cycles of 95°C for 1s and 60°C for 1 min. As a positive control bisulfite-converted DNA of DU145 and LNCap were used. Blank reactions with destillated water, which replaced DNA, served as negative control (NTC).

Eligibility criteria: Criteria for patients with prostate adenocarcinoma:

Inclusion Criteria

* sex: male
* diagnosis: prostate adenocarcinoma
* treatment: radical prostatovesiculectomy
* period of treatment: 11/30/2011 - 10/15/2013

Exclusion Criteria

* sex: female
* diagnosis: no prostate adenocarcinoma
* treatment: no radical prostatovesiculectomy
* period of treament: before 11/30/2011 or after 10/15/2013

Criteria for prostate adenocarcinoma negative control group:

Inclusion Criteria

* sex: male
* diagnosis: urothelial carcinoma
* treatment: cysto-prostatectomy
* period of treament: 12/14/2011 - 02/18/2014

Exclusion Criteria

* sex: female
* diagnosis: incidental prostate adenocarcinoma
* treatment: no cysto-prostatectomy
* period of treatment: before 12/14/2011 or after 02/18/2014

Primary outcomes:
measure: Methylation status of GSTP1
timeFrame: 2 years
measure: Histopathology of prostate fossa biopsies
timeFrame: 2 years
Secondary outcomes:
Other outcomes:
Locations:
city: Gronau
country: Germany
facility: St. Antonius Hospital
state: Nordrhein-Westfalen
city: Magdeburg
country: Germany
facility: Department of Urology, University Clinic Otto von Guericke University Magdeburg
state: Sachsen-Anhalt
Study officials:
affiliation: University of Magdeburg
name: Martin Schostak, Prof.Dr.med.
role: STUDY_CHAIR
References: