Detection of Tumor Genetic Alterations of Bladder Carcinomas in Body Fluids Depends on Sample Treatment before DNA Isolation

Utting, Michael; Müller, G.; Werner, Wolfram; Schubert, Jörg; Junker, Kerstin

doi:10.1111/j.1749-6632.2000.tb06593.x

Cited by 8 publications

(2 citation statements)

References 5 publications

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“…Plasma (contained EDTA), serum, and urine samples were collected preoperatively and stored frozen at −80°C, as well as tissue samples. After defrosting, body fluid samples were centrifuged at 15,000 g for 30 min 11. For DNA analysis, we used the supernatants only.…”

Section: Methodsmentioning

confidence: 99%

A Possible Noninvasive Method for the Detection of Bladder Cancer in Patients

Utting

Werner

Müller

et al. 2001

Annals of the New York Academy of Sciences

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Six microsatellite markers were selected to detect shifts or loss of heterozygosity (LOH) in urine, serum, and plasma samples of 44 bladder cancer patients. After centrifugation at 15,000g, we used supernatants for DNA analysis only. Tumor specimens were obtained by transurethral resection (TUR). Genetic alterations were detected in 33 of the 44 bladder tumors (75%). After polymerase chain reaction (PCR), DNA was detectable in 96% of all body fluid samples. Twenty-six percent of the detected microsatellite alterations of free DNA were tumor-specific, but 82% of all microsatellite changes of the tumors could be detected in body fluids. The study indicates that the simultaneous and multiple investigations of highly specific microsatellite markers could have a clinical relevance as a noninvasive tool for diagnosis and screening of bladder cancer. However, new ways for the sensitive DNA isolation of body fluids are needed.

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Section: Methodsmentioning

confidence: 99%

A Possible Noninvasive Method for the Detection of Bladder Cancer in Patients

Utting

Werner

Müller

et al. 2001

Annals of the New York Academy of Sciences

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“…Interestingly, the authors demonstrated that the examination of urine (from the paper it is not clear whether the cellular sediment or cell-free urine supernatant was used) is better suited than serum (52 LOH + MA in urine vs. 23 LOH + MA in serum). That the analysis of cell-free urine is better suited for microsatellite alterations than urine sediments was shown by several groups and the reason for this difference might be a "contamination" of the sediment with normal epithelial cells leading to a high noise vs. signal ratio [189,190]. The total amount of cell-free DNA and the relative quantity of long DNA (measured as 400 bp amplicons in q PCR) in urine supernatant is seen as a potential biomarker for bladder cancer [191].…”

Section: Cfnas In Stool and Urine (See Also Sect 4)mentioning

confidence: 98%

Extracellular Nucleic Acids and Cancer

Fleischhacker¹,

Schmidt²

2014

Advances in Predictive, Preventive and Personalised Medicine

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Since the clear proof of the presence of tumor-associated genetic alterations in extracellular nucleic acids almost 20 years ago this field gained has attracted much interest. According to our current knowledge it seems as if all tumor-associated alterations found in tumor cells are also found in the extracellular environment. The isolation of extracellular nucleic acids from tumor patients and its genetic characterization with very sensitive and highly specific methods led to the concept of "liquid biopsy". This means that for follow-up analysis of tumor patients, the physicians no longer depend exclusively on a single examination of tissue biopsies (usually at the time of diagnosis) but are able by longitudinally analyzing the extracellular nucleic acids to follow the reaction of a tumor to e.g. a given therapy, the development of resistance mechanisms. In this chapter we will discuss the detection and characterization of different genetic (e.g. mutation analysis and structural variations as seen in microsatellites), epigenetic (e.g. hypermethylation of selected sequences) and regulatory alterations (as in different miRNA expression patterns found in tumor patients). We will also touch on some confounding factors that have to be taken into consideration as well as the functional and biological aspects of extracellular nucleic acids.

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Serum DNA and urine DNA alterations of urinary transitional cell bladder carcinoma detected by fluorescent microsatellite analysis

et al. 2001

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There are no reliable serologic tumor markers for transitional cell carcinoma (TCC) of the urinary bladder and noninvasive urine investigations are inadequate. We used fluorescent microsatellite analysis (MSA) to detect serum DNA and urine-sediment DNA alterations in patients with bladder cancer and prospectively collected fresh tumor, peripheral blood, serum and spontaneous urine specimens from 39 consecutive patients treated for TCC of the bladder to obtain the corresponding DNA. Urinary bladder cancer accounts for approximately 3% of all newly diagnosed malignancies in Western countries. In contrast to most other tumors, bladder carcinoma offers the opportunity of noninvasive diagnosis via urinary cytology. Yet a reliable serologic tumor marker is not available. Follow-up for bladder carcinoma is demanding and time consuming, as most bladder cancers at diagnosis grow superficially and will recur in 70% of cases within 2 years of primary treatment. 1 The intensive follow-up for transitional cell carcinoma (TCC) of the bladder still relies on invasive diagnostic measures such as cystoscopy and bladder washings for urine cytology. Cytology has a low sensitivity in the diagnosis of well-differentiated papillary tumors. A reliable serologic marker and a highly sensitive urinary investigation could reduce the requirement and frequency of invasive procedures during follow-up.In recent studies molecular techniques have proved applicable for the detection of smallest amounts of free circulating tumor DNA in serum and plasma of cancer patients. 2,3 Using microsatellite analysis (MSA) Goessl et al. 3 identified plasma DNA alterations in 63% of patients with renal cell carcinoma. In recent studies MSA was also used to detect tumor-specific DNA alterations in urine sediment samples of patients with TCC of the urinary bladder. 4,5 In a pilot study Steiner et al. 4 achieved a sensitivity of 90% for the urinary cancer diagnosis with radiolabeled microsatellite markers. In contrast to cytology, this method is independent of tumor histomorphologic features, and thus offers high sensitivity in well-differentiated tumors. This molecular genetic assay, not previously looked at in TCC, could reliably identify tumor-specific alterations in the serum of patients and its use in follow-up after radical surgical therapy could lead to early tumor recurrence identification. In the case of superficial bladder cancers, MSA may be helpful in diagnosing local recurrence without the need for invasive measures dependent on morphologic tumor features. We therefore used MSA with 17 highly polymorphic fluorescently labeled microsatellite markers from the chromosomal regions 5q, 8p, 9p, 9q, 13q, 14q, 17p, 17q and 20q to detect serum DNA and sediment-DNA alterations from spontaneous urine samples of 39 consecutive patients treated for TCC of the urinary bladder, to evaluate the sensitivity of the method in the serologic and urinary diagnosis of bladder TCC. MATERIAL AND METHODS Tumor, blood and urine samplingIn 1999, we prospectively collected preo...

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Detection of Tumor Genetic Alterations of Bladder Carcinomas in Body Fluids Depends on Sample Treatment before DNA Isolation

Cited by 8 publications

References 5 publications

A Possible Noninvasive Method for the Detection of Bladder Cancer in Patients

A Possible Noninvasive Method for the Detection of Bladder Cancer in Patients

Extracellular Nucleic Acids and Cancer

Serum DNA and urine DNA alterations of urinary transitional cell bladder carcinoma detected by fluorescent microsatellite analysis

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