Positive Carcinoembryonic Antigen mRNA Research Articles

Prediction of peritoneal recurrences of gastric cancer by qPCR analysis of peritoneal lavage fluids.

52 Background: Peritoneal lavage cytology has been regarded as the only reliable method detect free cancer cells in the abdominal cavity. However, there certainly are patients who experience peritoneal recurrences despite negative cytology. Methods: Using qRT-PCR analysis, mRNA levels of 9 candidate molecular markers were quantified in peritoneal lavage fluids of from 187 patients with gastric cancer and 30 benign disease controls. ROC curve analysis was conducted to compare accuracies of the candidate markers. The cut-off points were set at the highest values of the benign disease controls. We evaluated predictive values of molecular markers in peritoneal lavage fluids for peritoneal recurrence. Results: From the result of ROC curve analysis, synaptotagmin XIII ( SYT13) and carcinoembryonic antigen ( CEA) mRNA levels were identified as predictive markers for peritoneal recurrences (The AUC value, 0.771, 0.775, respectively). 162 patients who had no concomitant peritoneal metastasis and were negative for lavage cytology (P0CY0), 13 patients (8.2%) had peritoneal recurrences. Those with either positive SYT13 or CEA mRNA level in peritoneal lavage fluids had significantly shorter peritoneal recurrence-free survival times compared to those with negative (3-years peritoneal recurrence-free survival rates, 69.5% vs 95.6%; P = 0.0069 and 79.3% vs. 93.4%; P = 0.0211, respectively). By combination of SYT13 and CEA, patients with both markers-positive had the greatest prevalence of peritoneal recurrences. Moreover, the peritoneal recurrence free survival curves of the both markers-positive group approached to that of the positive for lavage cytology group. Univariate analysis revealed that both SYT13-positive and CEA-positive were prognostic factors for peritoneal recurrence (hazard ratio: 4.21 and 3.53, respectively). In the multivariable analysis, SYT13-positive was found to be statistically significant independent prognostic factors (hazard ratio: 3.84, 95% confidence interval 1.24-13.2, P = 0.0202). Conclusions: SYT13 and CEA levels in peritoneal lavage fluids accurately predict peritoneal recurrences in patients with P0CY0.

Significance of carcinoembryonic antigen mRNA in peritoneal lavage determined by transcription–reverse transcription concerted method in patients with low rectal cancer

Positive carcinoembryonic antigen (CEA) messenger RNA (mRNA) in peritoneal lavage is associated with poor prognosis in patients with colon cancer. However, there are no reports about rectal cancer. Therefore we aimed to evaluate the frequency of positive CEA mRNA in peritoneal lavage and the significance of CEA mRNA in patients with low rectal cancer. A total of 55 patients with low rectal cancer who received curative surgical resection were enrolled. CEA mRNA in peritoneal lavage was measured using the transcription-reverse transcription concerted method, a quantitative RNA amplification method. The correlation between CEA mRNA and overall and peritoneal recurrence-free survival was evaluated. Among 55 patients, 6 (10.9%) had positive CEA mRNA in peritoneal lavage. Patients with positive CEA mRNA resulted in significantly higher recurrence rate than those with negative CEA mRNA (p=0.007). Similarly, the local recurrence rate was significantly higher in the positive CEA mRNA group than in the negative CEA mRNA group (p=0.0009). Lymph node metastasis and positive CEA mRNA were independent risk factors for overall and local recurrence. Positive CEA mRNA in low rectal cancer is a factor that predisposes patients to a high risk for overall recurrence, especially for local recurrence.

Detection of carcinoembryonic antigen mRNA in peritoneal lavage by the transcription-reverse transcription concerted method indicates poor prognosis in patients with stage II and III colon cancer

Peritoneal dissemination and positive peritoneal lavage cytology are associated with poor prognosis in colorectal cancer. Carcinoembryonic antigen (CEA) messenger RNA (mRNA) is often used as a marker to detect micrometastases. We aimed to evaluate the prognostic significance of CEA mRNA in the peritoneal lavage of colon cancer patients. Colon cancer patients (n = 201) who underwent curative operative resection between August 2009 and February 2013 were enrolled. CEA mRNA in peritoneal lavage was measured using the transcription-reverse transcription concerted method, a quantitative RNA amplification method. The correlation between CEA mRNA and overall and peritoneal recurrence-free survival was evaluated. Positive CEA mRNA in peritoneal lavage was an independent risk factor for overall recurrence-free survival in colon cancer (P < .0001). Positive CEA mRNA was a risk factor for poorer overall recurrence in stage II and III patients (P = .04 and P = .02, respectively). Moreover, among stage III patients with positive CEA mRNA, the postoperative chemotherapy group had significantly lower overall and peritoneal recurrence rates than the no postoperative chemotherapy group (P = .001). Positive CEA mRNA in peritoneal lavage was associated with high overall recurrence rates in stage II and III colon cancer. Further study is necessary to determinate the efficacy of aggressive postoperative chemotherapy for stage II and III colon cancer patients with positive CEA mRNA.

Effect of surgical manipulation on the dissemination of cancer cells into peripheral blood in patients with gastric cancer and its risk factor analysis

To evaluate the effect of surgical manipulation on the dissemination of cancer cells into blood circulation in patients with gastric cancer and to analyze its risk factors. This study included 45 consecutive patients with gastric cancer undergoing curative resection and 13 control cases (10 healthy persons and 3 patients with peptic ulcer receiving gastrectomy). Peripheral blood was obtained preoperatively and just after surgical manipulation. The mRNA levels of carcinoembryonic antigen (CEA) from the blood samples were assayed by reverse transcription-polymerase chain reaction(RT-PCR) and compared between the 2 groups. CEA mRNA was negative in all control cases. Of the 45 gastric cancer patients, the preoperative positive rate of CEA mRNA was 8.9%, while the postoperative positive rate was 48.9%, which was significantly higher than that of preoperation (P=0.000). Multivariable Logistic regression analysis showed that operative duration (P=0.014) and tumor depth (P=0.010) were independent risk factors for cancer cell dissemination. Furthermore, the operative duration in patients with positive postoperative CEA mRNA was markedly longer than that in patients with negative postoperative CEA mRNA (P=0.000), and positive rate of postoperative CEA mRNA in advanced gastric cancer was higher compared with that in early gastric cancer (P=0.034). Surgical manipulation of curative gastrectomy can provoke dissemination of cancer cells into blood circulation, and the operative duration and tumor invasion depth may be 2 of the risk factors for cancer cell dissemination.

Detection of Carcinoembryonic Antigen Messenger RNA-Expressing Cells in Peripheral Blood 7 Days After Curative Surgery is a Novel Prognostic Factor in Colorectal Cancer

The significance of detection of circulating cancer cells in blood during surgery in patients with colorectal cancer (CRC) remains controversial. Experimental study revealed that the cancer cells injected from the vein disappeared completely until 7 days. The aim of this study was to clarify that the detection of circulating cancer cells in blood taken later than 7 days after curative surgery may be a prognostic factor. Two hundred consecutive patients with CRC who underwent potentially curative surgery were the subjects. Peripheral blood was collected between 7 and 10 days after resection. Cancer cells were detected using reverse transcriptase-polymerase chain reaction targeting carcinoembryonic antigen (CEA) messenger RNA (mRNA). The median follow-up period was 52 months (range: 34-69 months). The overall positive incidence of CEA mRNA was 22%. Detection of CEA mRNA was not significantly related to conventional clinicopathological findings. Recurrence has been confirmed in 55 patients (28%). The recurrence rate was significantly higher in patients with rectal cancer, deep penetration, lymph node metastasis, preoperative chemoradiotherapy and positive CEA mRNA. The CEA mRNA positive patients showed significantly poorer disease free survival (DFS) and overall survival (OS) than the negative patients (DFS, P = 0.007; OS, P = 0.04). Multivariate analysis revealed that the positive expression of CEA mRNA (P < 0.01) as well as the tumor location and TNM stage classification was identified as the significant risk factors for recurrence. Detection of CEA mRNA expressing cells in peripheral blood 7 days after curative surgery is a novel independent factor predicting recurrence in patients with CRC.

Molecular Detection of Circulating Tumor Cells in the Peripheral Blood of Patients with Colorectal Cancer Using RT‐PCR: Significance of the Prediction of Postoperative Metastasis

Approximately 20%-45% of colorectal cancer (CRC) patients ultimately develop local recurrence or metastasis following curative surgical resection. The latter is caused by tumor cells shed from the primary carcinoma prior to or during operation, currently undetected by standard clinical staging. Fortunately, the presence of tumor cells in peripheral blood can be detected by molecular methods and is being regarded increasingly as a clinically relevant prognostic factor. To detect the presence of circulating tumor cells and evaluate their relationship to postoperative metastatic relapse, we simultaneously examined human telomerase reverse transcriptase (hTERT), cytokeratin-19 (CK-19), cytokeratin-20 (CK-20), and carcinoembryonic antigen (CEA) mRNA (messenger RNA) in the peripheral blood of 72 CRC patients and 30 healthy individuals. Using a reverse-transcriptase polymerase chain reaction (RT-PCR), these tumor-related mRNAs were amplified; in addition, analyses were carried out for their correlation with patients' clinicopathologic features, as well as the occurrence of postoperative metastasis. In RT-PCR analysis of the peripheral blood, 69.4% (50 out of 72), 66.7% (48 out of 72), 52.8% (38 out of 72), and 72.2% (52 out of 72) of CRC patients were positive for hTERT, CK-19, CK-20, and CEA mRNA respectively. All 30 healthy individuals were negative for hTERT and CEA mRNA expression, while 2 were positive for either CK-19 mRNA or CK-20 mRNA expression. The detection of CEA mRNA was significantly correlated with depth of tumor invasion (P=0.012), vessel invasion (P=0.035), TNM stage (P<0.0001), and postoperative metastasis (P<0.0001), while positive hTERT mRNA was correlated with TNM stage (P=0.037) and CK-19 was correlated with depth of tumor invasion (P=0.039) and postoperative metastasis (P=0.017). In addition, multivariate logistic regression showed that only CEA mRNA was an independent and significant predictor of postoperative metastasis (P=0.006). Our findings suggest that CEA mRNA may be a more reliable marker than hTERT, CK-19, and CK-20 for the detection of circulating cancer cells in the peripheral blood of CRC patients. Using RT-PCR for the detection of CEA mRNA is feasible and may be a promising tool for early detection of micrometastatic circulating tumor cells in CRC patients. CRC patients expressing positive CEA mRNA in peripheral blood have a significantly higher risk of postoperative metastasis. Nevertheless, confirmation of CEA mRNA as a prognostic predictive factor requires the continuation of patient follow-up.

Molecular Detection of Disseminated Tumor Cells in the Peripheral Blood of Patients with Gastric Cancer: Evaluation of Their Prognostic Significance

Early detection of disseminated tumor cells in the peripheral blood of patients with early stage gastric cancer could help to improve the outcome after tumor resection. The aim of this study is to evaluate the prognostic significance of tumor-related mRNA for the detection of circulating tumor cells in gastric cancer patients by a reverse-transcriptase polymerase chain reaction (RT-PCR) method. We simultaneously analyzed human telomerase reverse transcriptase (hTERT), cytokeratin-19 (CK-19), cytokeratin-20 (CK-20) and carcinoembryonic antigen (CEA) mRNA (messenger RNA) expression in the peripheral blood of 42 gastric cancer patients and 30 healthy individuals. Additionally, analyses were carried out for the correlation of these four molecular markers with patients’ clinicopathologic features, as well as the occurrence of postoperative recurrence/metastasis. Among 42 gastric cancer patients, the prevalence of mRNA for hTERT, CK-19, CK-20, and CEA was 61.9% (26/42), 69% (29/42), 61.9% (26/42), and 78.6% (33/42), respectively. All 30 healthy individuals were negative for hTERT and CEA mRNA, while two were positive for either CK-19 mRNA or CK-20 mRNA. Positive CEA mRNA was significantly correlated with tumor size (p = 0.008), vessel invasion (p = 0.001), depth of tumor invasion (p = 0.007), lymph node metastasis (p < 0.001), and TNM stage (p < 0.001). In addition, the multivariate logistic regression demonstrated that CEA mRNA expression was an independent and significant predictor for postoperative recurrence/metastasis (p = 0.032). Our findings suggest that CEA mRNA may be a more reliable marker than hTERT, CK-19 and CK-20 for the detection of circulating cancer cells in gastric cancer patients' peripheral blood. Patients with positive CEA mRNA expression in peripheral blood have a significantly higher risk of postoperative recurrence/metastasis.

Prognostic significance of CEA levels and detection of CEA mRNA in draining venous blood in patients with colorectal cancer

The aims of this study were to determine carcinoembryonic antigen (CEA) levels and incidence of tumor cells using the RT-PCR technique in draining venous blood of patients with colorectal cancer, correlate the results with various histopathologic factors and determine their significance as prognostic factors. From 1995 to 2000, 108 patients with adenocarcinoma of the colon or rectum, underwent curative surgery and enrolled in this prospective study. The 5-year survival group had significantly lower portal CEA levels compared to the hepatic metastasis outcome group. CEA mRNA was positive in the draining venous blood from 12 (11.1%) out of 108 patients included in the study. The rate of positive tumor cell detection in portal blood was significantly higher in the hepatic metastasis outcome group than in the 5-year survival and recurrence group. The proportion of patients with portal CEA > or =5 ng/ml was greater in patients with higher stage than in patients with lower stage. Positive CEA mRNA in draining venous blood predicted hepatic metastases and local recurrence with accuracy over 80% but with low sensitivity of 30% and 9%, respectively. Moreover, CEA level was a sensitive indicator in hepatic metastases as sensitivity was 95% and a specific indicator in predicting 5-year survival with specificity 84%.

Prospective validation of quantitative CEA mRNA detection in peritoneal washes in gastric carcinoma patients

Prediction of peritoneal relapse is extremely important for gastric cancer patients after curative surgery. The present study prospectively validates the prognostic ability of quantifying carcinoembryonic antigen (CEA) mRNA in peritoneal washes by real-time reverse transcriptase–polymerase chain reaction. Based on a retrospective study of 197 curatively resected gastric cancer patients (training set), we determined a cutoff value of CEA mRNA using receiver-operating characteristic curve. We used this cutoff value to validate the risk of peritoneal recurrence in a new cohort of 86 gastric cancer patients (validation set) between July 2000 and December 2002 in a prospective study. During the median 30 months of postoperative surveillance, 20 of the 86 patients died, and 13 of the 20 developed peritoneal metastases. Peritoneal recurrence-free survival as well as overall survival was significantly worse in patients with positive CEA mRNA (P<0.0001). Multivariate analysis with the Cox proportional hazards model showed that positive CEA mRNA was a significant independent risk factor with both survival (P=0.0130) and peritoneal recurrence-free survival (P=0.0006) as end points. These results indicate that quantitation of CEA mRNA in peritoneal washes is a reliable prognostic indicator of peritoneal recurrence in the clinical setting.

Quantitative detection of CEA expressing free tumor cells in the peripheral blood of colorectal cancer patients during surgery with real-time RT-PCR on a LightCycler

We applied novel real-time reverse transcriptase-polymerase chain reaction (RT-PCR) with a LightCycler for quantitative detection of carcinoembryonic antigen (CEA) mRNA expressing tumor cells in the peripheral blood of colorectal cancer patients. Analysis of peripheral blood samples from 99 potentially curative colorectal cancer patients revealed a significantly higher mean CEA mRNA value in post-operative bloods (18.71) than in pre-operative blood (1.03) (P=0.003). Kaplan–Meier analysis demonstrated disease free survival of patients with positive CEA mRNA in post-operative blood to be significantly shorter than in cases negative for CEA mRNA (P=0.03). These results suggest that tumor cells could be shed into the bloodstream during surgical procedures, and these free tumor cells are accompanied by a poor patient outcome. Real-time quantitative RT-PCR is a useful technique for quantitative assessment of free tumor cells in the peripheral blood of colorectal cancer patients.

Detection of circulating tumor cells by reverse transcriptase-polymerase chain reaction in patients with resectable non-small-cell lung cancer

Objective: We tested whether circulating tumor cells can be detected in the peripheral blood of patients with resectable non-small-cell lung cancer (NSCLC) by reverse transcriptase-polymerase chain reaction (RT-PCR) of carcinoembryonic antigen (CEA) mRNA. Methods: We assayed for CEA mRNA by RT-PCR in the peripheral blood, taken at the time of diagnosis before surgical intervention and again 2 to 3 weeks postoperatively, from 103 patients with NSCLC who underwent curative lobectomy. Blood samples taken from 15 patients with interstitial pulmonary fibrosis who underwent an open-lung biopsy and from 32 healthy subjects served as controls. Results: No control samples were positive for CEA by RT-PCR. Sixty-two (60%) of the preoperative blood samples from the 103 patients with NSCLC were positive. Of these 62 samples, 27 (44%) remained positive even after surgical intervention, whereas the remaining 35 samples (56%) became negative. The incidence of positive CEA mRNA correlated highly with pathologic TNM stage of disease in both the preoperative and postoperative blood samples. Conclusions: Many patients with resectable NSCLC have detectable levels of circulating cells expressing carcinoembryonic antigen even after surgical intervention. Such patients may have a higher rate of relapse. (Surgery 1999;126:820-6.)

Detection of circulating tumor cells by reverse transcriptase-polymerase chain reaction in patients with resectable non-small-cell lung cancer

We tested whether circulating tumor cells can be detected in the peripheral blood of patients with resectable non-small-cell lung cancer (NSCLC) by reverse transcriptase-polymerase chain reaction (RT-PCR) of carcinoembryonic antigen (CEA) mRNA. We assayed for CEA mRNA by RT-PCR in the peripheral blood, taken at the time of diagnosis before surgical intervention and again 2 to 3 weeks postoperatively, from 103 patients with NSCLC who underwent curative lobectomy. Blood samples taken from 15 patients with interstitial pulmonary fibrosis who underwent an open-lung biopsy and from 32 healthy subjects served as controls. No control samples were positive for CEA by RT-PCR. Sixty-two (60%) of the preoperative blood samples from the 103 patients with NSCLC were positive. Of these 62 samples, 27 (44%) remained positive even after surgical intervention, whereas the remaining 35 samples (56%) became negative. The incidence of positive CEA mRNA correlated highly with pathologic TNM stage of disease in both the preoperative and postoperative blood samples. Many patients with resectable NSCLC have detectable levels of circulating cells expressing carcinoembryonic antigen even after surgical intervention. Such patients may have a higher rate of relapse.