Guanylyl Cyclase C Messenger RNA Is a Biomarker for Recurrent Stage II Colorectal Cancer

Cagir, Burt; Gelmann, Alyssa; Park, Jason; Fava, Tracy A.; Tankelevitch, Alexander; Bittner, Edwin W.; Weaver, Eric J.; Palazzo, Juan; Weinberg, David S.; Fry, Robert D.; Waldman, Scott A.

doi:10.7326/0003-4819-131-11-199912070-00002

Cited by 96 publications

(101 citation statements)

References 26 publications

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“…RNA was extracted from tissues by a modification of the acid guanidinium thiocyanate-phenol-chloroform extraction method (19,23). Briefly, individual tissues were pulverized in 1.0 mL Tri-Reagent (Molecular Research Center, Cincinnati, OH) with 12 to 14 sterile 2.5-mm zirconium beads in a bead mill (Biospec, Bartlesville, OK) for 1 to 2 minutes.…”

Section: Methodsmentioning

confidence: 99%

“…Thus, GCC may fulfill the criteria for a sensitive and specific prognostic and predictive marker for identifying occult colorectal micrometastases (12). Indeed, in a retrospective analysis, detection of GCC expression by qualitative RT-PCR in lymph nodes free of metastases by histopathology was highly associated with the development of recurrent colorectal cancer (23).…”

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confidence: 99%

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A Validated Quantitative Assay to Detect Occult Micrometastases by Reverse Transcriptase-Polymerase Chain Reaction of Guanylyl Cyclase C in Patients with Colorectal Cancer

Schulz¹,

Hyslop²,

Haaf³

et al. 2006

Clinical Cancer Research

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Purpose: Guanylyl cyclase C (GCC), a receptor for bacterial diarrheagenic enterotoxins, may be a prognostic and predictive marker to detect occult micrometastases in patients undergoing staging for colorectal cancer. However, quantification of GCC expression in tissues by the quantitative reverse transcription-PCR (qRT-PCR) has not undergone analytic and clinicopathologic validation. Experimental Design: A technique to quantify GCC mRNA in tissues employing RT-PCR was developed and validated employing external calibration standards of RNA complementary to GCC.Results: GCC qRT-PCR exhibited reaction efficiencies >92%, coefficients of variations <5%, linearity >6 orders of magnitude, and a limit of quantification of >25 copies of GCC cRNA. This assay confirmed that GCC mRNA was overexpressed by colorectal tumors from 41 patients, which correlated with increased GCC protein quantified by immunohistochemistry. Analyses obtained with 164 lymph nodes from patients free of cancer and 15 nodes harboring metastases established a threshold for metastatic disease of f200 GCC mRNA copies/Ag total RNA, with a sensitivity of 93% and specificity of 97%. GCC mRNA above that threshold was detected in 76 of 367 (f21%) nodes free of disease by histopathology from 6 of 23 (26%) patients, suggesting the presence of occult micrometastases. Conclusions: Quantifying GCC mRNA in tissues by RT-PCR employing external calibration standards is analytically robust and reproducible, with high clinicopathologic sensitivity and specificity. This validated assay is being applied to f10,000 lymph nodes in a prospective trial to define the sensitivity of GCC qRT-PCR for staging patients with colorectal cancer.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

A Validated Quantitative Assay to Detect Occult Micrometastases by Reverse Transcriptase-Polymerase Chain Reaction of Guanylyl Cyclase C in Patients with Colorectal Cancer

Schulz¹,

Hyslop²,

Haaf³

et al. 2006

Clinical Cancer Research

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“…In intestine, GC-C is expressed along a crypt-to-villus gradient, with the greatest expression in the mid-villus where enterocytes transition from proliferation to differentiation, suggesting that GC-C may play a role in regulating that transition (22,23). Also, expression of GC-C is highly conserved, whereas that of guanylin is significantly reduced, in proliferating colorectal cancer cells and tumors (10,(24)(25)(26)(27). In addition, oral uroguanylin reduced the formation of polyps in the Min͞ϩ mouse model of colon cancer (27).…”

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confidence: 99%

Guanylyl cyclase C agonists regulate progression through the cell cycle of human colon carcinoma cells

Pitari

Guglielmo

Park

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

147

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“…This hypothesis is supported by a report by Rosenberg et al (2002), who found CK-positive cells on histological examination exclusively on the outside of lymph nodes in almost 30% of RT -PCR-positive specimens. Other markers, including CEA, MUC2, GCC, matrilysin, b-HCG and others, have been explored to detect lymph node micrometastases by RT -PCR and yielded inconsistent results (Ichikawa et al, 1998;Cagir et al, 1999;Bernini et al, 2000). Expression levels of these different markers can vary considerably among tumour specimens of the same entity.…”

Section: Discussionmentioning

confidence: 99%

CpG-island methylation of the ER promoter in colorectal cancer: analysis of micrometastases in lymph nodes from UICC stage I and II patients

et al. 2009

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Patients with UICC stage II colorectal cancer (CRC) have a risk of approximately 20% to develop disease recurrence after tumour resection. The presence and significance of micrometastases for locoregional recurrence in these patients lacking histopathological lymph node involvement on routine stained HE sections is undefined. Oestrogen receptor (ER) promoter methylation has earlier been identified in CRC. Therefore, we evaluated the methylation status of the ER promoter in lymph nodes from 49 patients with CRC UICC stage I and II as a molecular marker of micrometastases and predictor of local recurrence. DNA from 574 paraffinembedded lymph nodes was isolated and treated with bisulphite. For the detection of methylated ER promoter sequences, quantitative real-time methylation-specific PCR was used. Of the 49 patients tested, 15 (31%) had ER methylation-positive lymph nodes. Thirteen of those (86%) remained disease free and two (14%) developed local recurrence. In the resected lymph nodes of 34 of the 49 patients (69%), no ER promoter methylation could be detected and none of these patients experienced a local relapse. The methylation status of the ER promoter in lymph nodes of UICC stage I and II CRC patients may be a useful marker for the identification of patients at a high risk for local recurrence.

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Guanylyl Cyclase C Messenger RNA Is a Biomarker for Recurrent Stage II Colorectal Cancer

Abstract: Expression of guanylyl cyclase C mRNA in lymph nodes is associated with recurrence of colorectal cancer in patients with stage II disease. Analysis of guanylyl cyclase mRNA expression by RT-PCR may be useful for colorectal cancer staging.

Cited by 96 publications

References 26 publications

A Validated Quantitative Assay to Detect Occult Micrometastases by Reverse Transcriptase-Polymerase Chain Reaction of Guanylyl Cyclase C in Patients with Colorectal Cancer

A Validated Quantitative Assay to Detect Occult Micrometastases by Reverse Transcriptase-Polymerase Chain Reaction of Guanylyl Cyclase C in Patients with Colorectal Cancer

Guanylyl cyclase C agonists regulate progression through the cell cycle of human colon carcinoma cells

CpG-island methylation of the ER promoter in colorectal cancer: analysis of micrometastases in lymph nodes from UICC stage I and II patients

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