Laser Microdissection of Small Tissue Samples – Application to Chronic Pancreatitis Tissues

Heinmöller, Ernst; Bockholt, Anke; Werther, Meike; Ziemer, Maria; Müller, A. Hermann; Ghadimi, B. Michael; Rüschoff, Josef

doi:10.1078/0344-0338-00432

Cited by 9 publications

(6 citation statements)

References 32 publications

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“…Tissue fragments were collected in the adhesive lid of a microfuge tube. Immediately following LMD, tissue was removed from the lid and immersed in 100-ml lysis buffer (Macherey Nagel, Düren, Germany; Heinmoller et al 2003, Niyaz et al 2005.…”

Section: Laser Microdissectionmentioning

confidence: 99%

Microarray analysis reveals differential expression of benign and malignant pheochromocytoma

Waldmann

Fendrich²,

Holler³

et al. 2010

Endocrine-Related Cancer

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The diagnosis of a malignant pheochromocytoma (PC) can only be established by the presence of distant metastases, but a subset of apparently benign PCs develop metastases. We have employed a microarray analysis to identify a typical gene expression profile which distinguishes malignant from benign PC. Total RNA was isolated from fresh-frozen tissue of five benign and five malignant PCs. The reference consisted of laser microdissected tissue from normal adrenal medulla. After generating Cy3-and Cy5-fluorescently labeled cDNAs, F-chips containing 11 540 spots were hybridized. Data were analyzed with the IMAGENE 3.0 software. Gene expression levels were validated by real-time (RT)-PCR and immunohistochemistry (IHC). The analysis revealed a more than twofold difference in expression between benign and malignant PCs in 132 genes: 19 were up-regulated and 113 were down-regulated. Expression differences of six genes (calsequestrin, NNAT, neurogranin, secreted protein acidic and rich in cysteine (SPARC), EGR2, and MAOB) were confirmed by RT-PCR in 25 PCs. IHC for calsequestrin revealed an overexpression in malignant PCs (7/10 vs 1/10, PZ0.03). Comparative analysis by microarray of all ten PCs (benign/malignant) versus normal adrenal medulla revealed a more than twofold expression difference in 455/539 and 491/671 genes respectively. Several of these genes are known to participate on adrenal tumorigenesis, potential tumor suppressor genes, and oncogenes. Comprehensive gene expression analysis of malignant and benign PCs revealed different gene profiles, which could be used to discriminate between malignant and benign PCs. Based on these findings, the strategy for further follow-up and treatment could be modified accordingly.

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Section: Laser Microdissectionmentioning

confidence: 99%

Microarray analysis reveals differential expression of benign and malignant pheochromocytoma

Waldmann

Fendrich²,

Holler³

et al. 2010

Endocrine-Related Cancer

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“…The presence of other undetected cancer in other organs cannot be totally excluded, although all patients were more thoroughly examined during their hospital stay than the 20 healthy individuals of similar age and without detectable LOH. In a recent study, Heinmo¨ller et al 19 reported rare LOH in obstructive duct lesions of the chronic inflamed pancreas and suggested that a chronic hyperproliferative stimulus may lead to increased cellular turnover and to accumulation of mutations over time. However, this study was performed by laser microdissection of pancreatic intraductal lesions in tissues of chronic pancreatitis.…”

Section: Discussionmentioning

confidence: 97%

Microsatellite Analysis in Serum DNA as a Diagnostic Tool for Distinction of Patients With Unknown Pancreatic Masses

Wachowiak

Kaifi

Schwarzenbach

et al. 2007

Diagnostic Molecular Pathology

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The clinical distinction between cancer and chronic pancreatitis is difficult in patients with pancreatic masses. To test whether detection of aberrant serum DNA could assist in this important differential diagnosis, we tested a panel of 12 microsatellitemarkers from chromosomes 17p, 17q, 13q, 9p, 5q, and 2p in the blood of 35 pancreatic cancer patients, 22 patients with chronic pancreatitis, and 20 healthy individuals. An average of 2.8 loss of heterozygosity (LOH) was found in 32 of 35 cancer patients of whom 30 (86%) had 2 or more LOH. LOH was also found in 7 of 22 pancreatitis patients but all these patients had only 1 LOH. No LOH was detected in healthy donors of comparable age. These data suggest that LOH analysis may be a substantial help for diagnosing pancreatic masses. An extension of the panel, perhaps in combination with a better selection of markers may further improve this assay.

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“…The LMD, as we have described above, is a preparative method, since it guarantees the recovery of distinct cell populations. For this reason, quantitative PCR appears to be by far the most used technique, that is frequently used in combination with the LMD in the pancreatic cancer studies [40,41], in chronic pancreatitis and preneoplastic lesions [42], since the expression levels of RNA of specific molecular determinants can be compared in different cell types present within a sample [23,24,28]. By this procedure it is possible to separate and study different cell populations present in the same tissue sample.…”

Section: Lmd On Primary Cell Culturesmentioning

confidence: 99%

Advances in Primary Cell Culture of Pancreatic Cancer

Funel

2014

Molecular Diagnostics and Treatment of Pancreatic Cancer

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Laser Microdissection of Small Tissue Samples – Application to Chronic Pancreatitis Tissues

Cited by 9 publications

References 32 publications

Microarray analysis reveals differential expression of benign and malignant pheochromocytoma

Microarray analysis reveals differential expression of benign and malignant pheochromocytoma

Microsatellite Analysis in Serum DNA as a Diagnostic Tool for Distinction of Patients With Unknown Pancreatic Masses

Advances in Primary Cell Culture of Pancreatic Cancer

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