AgarCyto: A Novel Cell-processing Method for Multiple Molecular Diagnostic Analyses of the Uterine Cervix

Kerstens, Harold M.J.; Robben, Joris; Poddighe, Pino J.; Melchers, Willem J. G.; Boonstra, H.; Wilde, P.C.M. de; Macville, Merryn; Hanselaar, A. G. J. M.

doi:10.1177/002215540004800515

Cited by 58 publications

(62 citation statements)

References 24 publications

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“…For prolactin receptor immunostaining, cells were collected and cell pellets were embedded in bacto agar (Becton Dickinson and Company, Franklin Lakes, NJ, USA), fixed in formalin, and embedded in paraffin following standard procedures as published earlier. 24 …”

Section: Cell Linesmentioning

confidence: 99%

Clinicopathological significance of prolactin receptor expression in colorectal carcinoma and corresponding metastases

et al. 2010

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The role of human prolactin and its receptor, the prolactin receptor, in colorectal cancer is largely unknown. Our study aimed to assess the prevalence of prolactin receptor expression, its association with clinicopathological variables, as well as its prognostic value, comparing results of primary tissues with those of corresponding metastases. In all, 373 primary colorectal cancer and 171 corresponding metastases were evaluated for prolactin receptor expression by immunohistochemistry using a tissue microarray technique. Immunoreactivity was semiquantitatively scored as either focal (o10% of tumor cells positive), moderate (10-50%), or extensive (450%). Prolactin receptor expression was related to clinicopathological parameters as well as patient outcome. To substantiate our findings, prolactin receptor expression was additionally assessed in HT-29 and SW-480 colorectal cancer cell lines using western blot. Prolactin receptor expression was observed in 360 out of 373 (97%) primary tumors, with 21 (6%) cases showing focal, 55 (15%) moderate, and 284 (76%) extensive expression, respectively. Extensive prolactin receptor expression was significantly associated with tumor size (P ¼ 0.002) and grade (Po0.001) as well as histological subtype (Po0.001). Somer's D coefficients for concordance of primary tumors with corresponding lymph node and distant metastases were D ¼ 0.719 (Po0.001) and D ¼ 0.535 (P ¼ 0.001), respectively. Extensive prolactin receptor expression was significantly associated with disease progression (P ¼ 0.03) and cancer-specific survival (P ¼ 0.04) in patients with highgrade cancers. In conclusion, prolactin receptor expression is common in colorectal cancer, with high concordance between primary tumors and corresponding metastases. In view of evolving targeted therapy concepts in colorectal cancer, widespread prolactin receptor expression may offer a therapeutic perspective in affected patients.

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Section: Cell Linesmentioning

confidence: 99%

Clinicopathological significance of prolactin receptor expression in colorectal carcinoma and corresponding metastases

et al. 2010

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“…All cell lines were obtained from the American Tissue and Cell Collection (Manassas, VA, USA) and were cultured according to the supplier's instructions. We used cell lines obtained either directly from cultures or after formalin-fixation and paraffin-embedding, as adapted from the published protocol by Kerstens et al 17 Surgically resected tissues were collected from 66 colorectal carcinoma patients (48 male and 18 female) at the Surgical Department of the Danube Hospital, Vienna, Austria. Median age at surgery was 67 years (range, 41 to 87 years).…”

Section: Controls and Patient Samplesmentioning

confidence: 99%

High Quality Assessment of DNA Methylation in Archival Tissues from Colorectal Cancer Patients Using Quantitative High-Resolution Melting Analysis

Balić

Pichler

Strutz

et al. 2009

The Journal of Molecular Diagnostics

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High-resolution melting (HRM) analysis is a novel tool for analysis of promoter methylation. The aim of the present study was to establish and validate HRM analysis for detection of promoter methylation on archival formalin-fixed paraffin-embedded tissues from colorectal cancer patients. We first evaluated HRM assays for O 6 -methylguanine-DNA methyltransferase (MGMT) and adenomatous polyposis coli (APC) promoter methylation on a methylated DNA dilution matrix and DNA extracted from eight fresh or formalin-fixed paraffin-embedded human cancer cell lines. Then we used these assays for the analysis of MGMT and APC promoter methylation in a subset of archival formalin-fixed paraffin-embedded colorectal tumor specimens. All samples with promoter methylation of MGMT or APC and randomly selected samples without promoter methylation were analyzed twice. All results generated by HRM were validated with MGMT and APC MethyLight assays. APC and MGMT promoter methylation data were consistent and reproducible throughout the dilutions and all three replicates in the methylated DNA dilution matrix and between two experiments in clinical samples. There was high concordance between HRM and MethyLight results. HRM for APC promoter methylation revealed consistent results between fresh and formalin-fixed paraffin-embedded human cancer cell line DNA. The methylation status in archival tumor specimens from patients with colorectal cancer can therefore be determined with high quality by HRM. The ability to analyze archival tissues greatly facilitates further research and its clinical implementation.

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“…Cell suspensions of human melanoma cell lines 1F6 and Mel57 were processed into AgarCyto's as previously described (Kerstens et al, 2000). 4 mm sections of paraffin-embedded AgarCyto's and melanocytic tumour lesions were mounted on Superfrost microscope slides.…”

Section: Immunohistochemistrymentioning

confidence: 99%

Analysis of differential gene expression in human melanocytic tumour lesions by custom made oligonucleotide arrays

et al. 2005

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Melanoma is one of the most aggressive types of cancer and resection of the tumour prior to dissemination of tumour cells is still the most effective treatment. Therefore, early diagnosis of melanocytic lesions is important and identification of novel (molecular) markers would be helpful to improve diagnosis. Moreover, better understanding of molecular targets involved in melanocytic tumorigenesis could possibly lead to development of novel interventions. In this study, we used a custom made oligonucleotide array containing 298 genes that were previously found to be differentially expressed in human melanoma cell lines 1F6 (rarely metastasising) and Mel57 (frequently metastasising). We determined differential gene expression in human common nevocellular nevus and melanoma metastasis lesions. By performing nine dye-swap array experiments, using individual as well as pooled melanocytic lesions, a constant differential expression could be detected for 25 genes in eight out of nine or nine out of nine array analyses. For at least nine of these genes, namely THBD, FABP7, H2AFJ, RRAGD, MYADM, HR, CKS2, NCK2 and GDF15, the differential expression found by array analyses could be verified by semiquantitative and/or real-time quantitative RT -PCR. The genes that we identified to be differentially expressed during melanoma progression could be potent targets for diagnostic, prognostic and/ or therapeutic interventions.

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AgarCyto: A Novel Cell-processing Method for Multiple Molecular Diagnostic Analyses of the Uterine Cervix

Cited by 58 publications

References 24 publications

Clinicopathological significance of prolactin receptor expression in colorectal carcinoma and corresponding metastases

Clinicopathological significance of prolactin receptor expression in colorectal carcinoma and corresponding metastases

High Quality Assessment of DNA Methylation in Archival Tissues from Colorectal Cancer Patients Using Quantitative High-Resolution Melting Analysis

Analysis of differential gene expression in human melanocytic tumour lesions by custom made oligonucleotide arrays

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