Multiple Molecular Markers as Predictors of Colorectal Cancer in Patients with Normal Perioperative Serum Carcinoembryonic Antigen Levels

Abstract

Purpose: In this study, a high-sensitivity colorimetric membrane array method was used to detect circulating tumor cells (CTC) in the peripheral blood of colorectal cancer (CRC) patients with normal perioperative serum carcinoembryonic antigen (CEA) levels. This membrane array method was evaluated as a potential diagnostic and postoperative surveillance tool. Study Design: Membrane arrays consisting of a panel of mRNA markers that include human telomerase reverse transcriptase, cytokeratin-19, cytokeratin-20, and CEA mRNA were used to detect CTCs in the peripheral blood of 157 postoperative CRC patients with normal perioperative serum CEA levels and in 80 healthy individuals. Digoxigenin-labeled cDNA were amplified by reverse transcription-PCR from the peripheral blood samples, which were then hybridized to the membrane array. The sensitivity, specificity, and accuracy of membrane arrays for the detection of CTCs were then calculated. Results: Using the four markers in combination, expression of any three markers or all the four markers in this panel was significantly correlated with the clinicopathologic characteristics, including depth of tumor invasion, lymph node metastasis, tumor-node-metastasis stage, and postoperative relapse (all P < 0.05). The interval between the detection of all four positive molecular markers and subsequent elevated CEA ranged from 3 to 8 months (median 6 months). The expression of all four mRNA markers was an independent predictor for postoperative relapse. CRC patients with all four mRNA markers expression showed a significantly poorer survival rate than those with less than four positive markers. Conclusions: The constructed membrane array method was helpful in the early prediction of postoperative relapse in CRC patients with normal perioperative serum CEA levels.

Prognostic significance of circulating tumour cells following surgical resection of colorectal cancers: a systematic review

“…Eight studies were excluded as they reported only on preoperative blood sampling (Bessa et al, 2001;Fujita et al, 2001;Wong et al, 2001;Zhang et al, 2005;Douard et al, 2006;Iinuma et al, 2006;Wang et al, 2006;Friederichs et al, 2007). Nine studies were excluded as they contained data from which outcomes could not be extracted, of which four were excluded as they combined positive results of molecular detection of multiple samples and their effect on prognosis (Hardingham et al, 2000;Guller et al, 2002;Bosch et al, 2003;Koyanagi et al, 2008), and five were excluded as it was impossible to extract the data for outcomes of interest (White and Griffiths, 1976;Funaki et al, 1998;Wyld et al, 1998;Conzelmann et al, 2005;Wang et al, 2007). Five studies were excluded as they were reviews or systematic reviews, which did not report on prognostic outcomes (Tsavellas et al, 2001;Sleijfer et al, 2007;Riethdorf et al, 2008;Sergeant et al, 2008;Tsouma et al, 2008).…”

“…The 14 studies included here comprised 1841 patients in total, with the median number of patients in each study being 99.5 (range 42 -438) (Taniguchi et al, 2000;Yamaguchi et al, 2000;Ito et al, 2002;Bessa et al, 2003;Chen et al, 2004;Sadahiro et al, 2005Sadahiro et al, , 2007Katsumata et al, 2006;Koch et al, 2006;Lloyd et al, 2006;Allen-Mersh et al, 2007;Uen et al, 2007Uen et al, , 2008Wang et al, 2007).…”

“…Of the 14 studies included, eight undertook blood sampling perioperatively (Taniguchi et al, 2000;Yamaguchi et al, 2000;Ito et al, 2002;Chen et al, 2004;Sadahiro et al, 2005;Katsumata et al, 2006;Koch et al, 2006;Lloyd et al, 2006), four undertook sampling approximately 24 h after resection (Bessa et al, 2003;Koch et al, 2006;Lloyd et al, 2006;Allen-Mersh et al, 2007), and six undertook sampling between 24 h and 14 days after resection (Chen et al, 2004; Allen-Mersh et al, 2007;Sadahiro et al, 2007;Uen et al, 2007Uen et al, , 2008Wang et al, 2007).…”

“…Seven investigated the role of CTC in patients with stage-I to stage-III disease undergoing potentially curative surgery (Ito et al, 2002;Bessa et al, 2003;Sadahiro et al, 2005Sadahiro et al, , 2007Allen-Mersh et al, 2007;Wang et al, 2007;Uen et al, 2008); three studied CTC in patients with early CRC (of which two investigated stage-II disease alone (Koch et al, 2006;Uen et al, 2007) and one investigated stage-I and stage-II disease (Lloyd et al, 2006)); and four papers included patients with stage-I to stage-IV disease (Taniguchi et al, 2000;Yamaguchi et al, 2000;Chen et al, 2004;Katsumata et al, 2006).…”

A prospective study comparing multi-gene biomarker chip and serum carcinoembryonic antigen in the postoperative surveillance for patients with stage I–III colorectal cancer

“…Reports have described the detection of circulating tumor cells (CTCs) in the peripheral blood of CRC patients; this method has major prognostic and therapeutic implications [7][8][9][10]. Our recently developed membrane array-based multigene biomarker assay can detect CTCs in the peripheral blood of CRC patients; this is a rational approach for the surveillance of postoperative CRC patients [6,[11][12][13][14][15]. However, a detailed prospective comparative study regarding the diagnostic accuracy of the biomarker chip and serum CEA level detection is required.…”