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Background: Carcinoembryonic antigen (CEA) detection was evaluated in breast cancer (BC). The statistical correlation between the CEA mRNA and clinico-pathological features in the peripheral blood (PB) and tissue samples of BC was assessed. Materials and Methods: RT-PCR (Reverse transcription-polymerase chain reaction) analysis was applied to study the expression of CEA in PB of 30 healthy females and 30 patients with operable BC before receiving any therapy, as well as in the tissue of 30 BC patients. Results: CEA was observed in a number of normal subjects, but there was a significant difference between the patients and controls. The detected CEA mRNA from tissue samples were the same as PB of patients and a correlation was observed between the CEA mRNA in PB and tissue samples (Pearson chi-square = 8.62, P=0.003). In the PB, CEA mRNA was significantly different in HER-2 (-)/HR (+) compare with HER-2(+)/HR (-) tumor group (p=0.026). Finally, CEA in serum was also significantly different in HER-2(-)/HR (+) compared with HER-2(+)/HR (+) and HER-2(+)/HR (-) subtypes (p=0.008 and p=0.043, respectively). Conclusion: CEA mRNA evaluation is diagnostically valuable as a breast cancer marker. Additionally, CEA can significantly improve the sensitivity of diagnosis.

Introduction: Cancer is one of the most important causes of mortality in the world. So, in this study the changes of expressing miR-205 and CEA in oral cancer in peripheral blood were examined for early detection and better treatment. Methods: In this study, we selected the number of 30 patient people and 30 healthy people. We measured their blood miR-205 and CEA using Real-Time PCR technique and evaluated the relationship between the expression of these biomarkers with tumor staging and cancer progression. Findings: there is no a significant difference in mean age by comparing these two groups using t-test. The CEA mRNA biomarker was positive in 24 out of 30 people of the patient people group and was positive in 4 out of 30 people of the healthy people group. Statistical comparison represented a statistically significant difference between the two groups (P-value

Introduction: Lung cancer is the most common cause of cancer-related death among males and females. The diagnosis of lung cancer is of great importance for clinical considerations and follow-up treatment. This study aimed to examine the expression of CEA, LUNX, and CK19 biomarkers in the cancerous and healthy tissues of patients suffering from NSCLC. Methods: In this study, 30 patients with NSCLCs referring to Masih Daneshvari Hospital in Tehran were voluntarily selected prior to taking any treatment. A tissue sample from the center and a sample of healthy tissues close to the cancerous masses were prepared by a specialist in the bronchoscopy sector and tested using real-time RT-PCR. Results: Positive CEA mRNA was observed in cancerous tissues in the center of tumors of 25 out of 30 cases. In the healthy tissue group, the same was found in 10 out of 30 cases (P

Background The tumor microenvironment (TME) is critical to every aspect of cancer biology. Organotypic tumor slice cultures (TSCs) preserve the original TME and have demonstrated utility in predicting drug sensitivity, but the association between clinicopathologic parameters and in vitro TSC behavior has not been well-defined. Methods One hundred and eight fresh tumor specimens from liver resections at a tertiary academic center were procured and precisely cut with a Vibratome to create 250 μm × 6 mm slices. These fixed-dimension TSCs were grown on polytetrafluoroethylene inserts, and their metabolic activities were determined by a colorimetric assay. Correlation between baseline activities and clinicopathologic parameters was assessed. Tissue CEA mRNA expression was determined by RNAseq. Results By standardizing the dimensions of a slice, we found that adjacent tumor slices have equivalent metabolic activities, while those derived from different tumors exhibit >30-fold range in baseline MTS absorbances, which correlated significantly with the percentage of tumor necrosis based on histologic assessment. Extending this to individual cancers, we were able to detect intra-tumoral heterogeneity over a span of a few millimeters, which reflects differences in tumor cell density and Ki-67 positivity. For colorectal cancers, tissue CEA expression based on RNAseq of tumor slices was found to correlate with clinical response to chemotherapies. Conclusions We report a standardized method to assess and compare human cancer growth ex vivo across a wide spectrum of tumor samples. TSC reflects the state of tumor behavior and heterogeneity, thus providing a simple approach to study of human cancers with an intact TME.

Background:Several prognostic factors were proposed to improve early detection of recurrence after liver resection of metastases of colorectal cancer. Circulating tumor cell-related transcripts were evaluated in colorectal cancer patients with conflicting results. The aim of this study was to investigate usefulness of carcinoembryonic antigen CAM5, epidermal growth factor receptor, and ERCC1 transcripts in the bloodstream as predictive factors of recurrence in patients who underwent liver resection for metastases of colorectal cancer.Methods:Peripheral blood was collected from 29 patients at the time of the colorectal cancer liver metastasis resection, and from 25 normal controls. Follow-up draws (FUDs) were also performed at 30 days, and 3 and 12 months since surgery. On each sample, carcinoembryonic antigen CAM5, ERCC1, and GAPDH mRNAs were examined by quantitative reverse transcription (qRT).Results:Carcinoembryonic antigen transcript levels were linearly correlated to the number of spiked cells (qRT analytical limit = five cells). Among 29 patients (20 M/9 F; mean age 63 years (range 32–79), highly significant levels of carcinoembryonic antigen, if compared to the baseline, were detected in those relapsing after surgery ( P Conclusions:Blood carcinoembryonic antigen-mRNA absolute copy number overtime variation can represent a valid early predictor of relapse after liver resection in colorectal liver metastases patients. Prospective studies, in the context of large clinical trials, will provide further data to also qualify ERCC1 as a predictive biomarker for decisions on therapeutic strategies.

OBJECTIVE: To assess the effect of hyperthermic intraperitoneal chemotherapy (HIPEC) to eradicate intraperitoneal free cancer cells and to explore the feasibility of cytological cure for peritoneal carcinomatosis (PC). METHODS: The peritoneal lavage fluid (or ascites) from 50 PC patients was collected before and after intraoperative HIPEC, respectively, for conventional cytology test, and conventional and real-time quantitative reverse transcript polymerase chain reaction detecting carcinoembryonic antigen (CEA) mRNA and cytokeratin-20 (CK20) mRNA. The blood samples 3 days before and 7 days after intraoperative HIPEC were also collected for detecting the serum tumor markers, including CEA, carbohydrate antigen (CA) 125, and CA19-9. RESULTS: The positive rate of conventional cytology test before HIPEC versus after HIPEC was100.0% versus 22.0% (P = .000). The positive rates of CEA mRNA and CK20 mRNA before HIPEC versus after HIPEC were 100.0% versus 86.0% (P = .012) and 100.0% versus 96.0% (P = .495), respectively. Moreover, after HIPEC, 18 (36.0%) patients had a decline in CEA mRNA (P = .000), and 17 (34.0%) patients had a decline in CK20 mRNA (P = .000). The positive rates of serum CEA, CA125, and CA199 before HIPEC versus after HIPEC were 52.0% versus 28.0% (P = .014), 52.0% versus 44.0% (P = .423), and 40.0% versus 28.0% (P = .205), respectively. CONCLUSION: HIPEC could effectively eradicate intraperitoneal free cancer cells and partially achieve cytological cure for PC.

Objective To explore the clinical significance of expression of CEA mRNA and serum CEA and the related proteins in colorectal cancer (CRC). Methods Blood samples were collected from 370 CRC patients and 350 controls. CEA mRNA was determined by RT-PCR and levels of CEA, CA19-9, CA242, and CA724 were examined with chemiluminescence. Results The positive rate of jointly detecting serum CEA, CA19-9, CA242, and CA724 was significantly higher than CEA mRNA expressions (P

e16090Background: Most patients have peritoneal metastasis over the course of gastric cancer. The prognosis of these patients with peritoneal metastasis is extremely poor. The aim of this study is ...

The current authors previously identified circulating cells expressing carcinoembryonic antigen (CEA) messenger ribonucleic acid (mRNA) in 80% of lung cancer patients bearing distant metastases. The current study prospectively validated the data on a novel cohort and extended the study to other mRNAs expressed by neoplastic cells. CEA, cytokeratin 19 and 20, aldolase A and epithelial glycoprotein 2 (EPG2) mRNA was analysed by reverse transcriptase-polymerase chain reaction in circulating cells from 19 healthy controls, and in biopsies and blood at diagnosis from 32 lung cancer patients monitored for 24 months. Aldolase A and cytokeratin 19 mRNA occurred in circulating cells of all controls; cytokeratin 20 was not expressed by any lung cancer biopsy. EPG2 mRNA occurred in all biopsies but not in the patients' circulating cells. CEA mRNA occurred in 29/32 (90.6%) biopsies and in 17/32 mRNA samples from circulating cells from lung cancer patients. Of these positive patients 12/17 developed metastases within 9 months of mRNA analysis. Three positive patients died, one was lost to follow-up, and one did not develop metastases within 24 months. Of the negative patients 12/15 did not develop metastases during the 24-month follow-up; one patient was lost to follow-up, one did not express CEA, and another developed metastases. Unlike in other neoplasias, cytokeratin 19 and 20, aldolase A and epithelial glycoprotein 2 messenger ribonucleic acid are not useful for the detection of circulating cancer cells in lung cancer. Carcinoembryonic antigen messenger ribonucleic acid analysis in circulating cells helps to identify lung cancer patients at a greater risk of metastases. Grants from Ministero Italiano dell'Universita e della Ricerca (MIUR; Rome, Italy); from the Consiglio Nazionale delle Ricerche (CNR) Targeted project “Biotecnologie”; and from Regione Campania (Ricerca Sanitaria Finalizzata and L.R. 41/96) are gratefully acknowledged.

初発大腸癌開腹手術症例42例を対象,良性疾患5例を対照とし術中腹腔洗浄液の細胞診及びRT-PCR法を用いたCEAm-RNAの定性を施行し,その臨床病理学的意義を検討した.CEAm-RNA陽性率は19.0%,細胞診陽性率は7.1%であった.良性疾患ではCEAm-RNA,細胞診共に陰性であった.肉眼的腹膜播種性転移陽性症例3例全例はCEAm-RNA,細胞診ともに陽性であった.CEAm-RNA陽性,細胞診陰性症例5例中1例に術後腹膜再発を認めた.RT-PCR法を用いたCEAm-RNAの定性は細胞診に比,癌細胞の存在診断をより鋭敏に可能とする可能性があり,有用な予後判定因子になり得ると考えられた.また臨床病理学的検討により,遊離癌細胞を生じやすい,あるいは合併しやすい因子として深達度SS以上で浸潤様式が散在性の症例,組織型が高分化腺癌よりも中分化腺癌,粘液癌の症例,リンパ節転移,リンパ管浸襲を有する症例,肝転移を有する症例,Stage II以上の症例が考えられた.

AIM To evaluate the clinical and prognostic significance of preoperative and postoperative cytokeratin 19 (CK19) and carcinoembryonic antigen (CEA) mRNA levels in peripheral blood of patients with gastric cardia cancer (GCC). METHODS We detected the preoperative and postoperative mRNA levels of CK19 and CEA in peripheral blood of 129 GCC patients by using reverse transcription-polymerase chain reaction and evaluated their clinical and prognostic significance by univariate Kaplan-Meier survival analysis and multivariate Cox proportional hazard analysis. A new prognostic model which stratified patients into three different risk groups was established based on the independent prognostic factors. RESULTS Elevated preoperative and postoperative CK19 and CEA mRNA levels in peripheral blood of GCC patients were associated with lymph node metastasis. Univariate analysis showed that tumor size, histological grade, depth of tumor invasion, lymph node metastasis, preoperative CK19 mRNA, and preoperative and postoperative CEA mRNA levels were correlated with the prognosis of GCC patients. The multivariate analysis showed that lymph node status (P = 0.018), preoperative CK19 (P = 0.035) and CEA (P = 0.011) mRNA levels were independent prognostic factors for overall survival (OS). The 5-year OS rates for the low-, intermediate-, and high-risk groups were 48.3%, 22.6%, and 4.6%, respectively (P < 0.001). CONCLUSION Elevated preoperative CK19 and CEA mRNA levels may be regarded as promising biomarkers for predicting lymph node metastasis and poor prognosis in patients with GCC. This new prognostic model may help us identify the subpopulations of GCC patients with the highest risk.

Background Carcinoembryonic antigen (CEA) mRNA expression in peritoneal washes from gastric cancer patients has been reported as an indicator for survival or peritoneal recurrence. The whole process of CEA mRNA detection is time- and labor-intensive. We report the potential of One-Step Nucleic acid Amplification (OSNA) as a rapid and simple system for CEA mRNA detection in peritoneal washes. Methods A total of 128 peritoneal washes were analyzed by cytological examination including immunocytochemistry. After the cytological examination, the CEA mRNA concentration in the residual cells was measured using the OSNA system. The CEA mRNA concentration in peritoneal washes was compared with the results of the cytological examination. Results CEA mRNA at concentrations from 10 to 107 copies/μL was detected by the OSNA system within 10 min, and an excellent correlation was observed between the logarithmic CEA mRNA concentration and the detection time (r = 0.998). The CEA mRNA cutoff value for distinguishing positive and negative cases through cytological examination was identified as 25 copies/μL. At this cutoff value, the concordance rate with the cytological examination was 93.8%. The overall survival in CEA mRNA-positive versus -negative cases identified using the OSNA system was statistically significant. Conclusion This CEA mRNA detection system shows potential for cancer cell detection and for routine use in the clinical laboratory because of its simplicity and rapidity.

We assessed whether circulating cell positivity using RT-PCR for carcinoembryonic antigen (CEA) cDNA was affected by venesection via a needle compared/with a pre-aspirated venous cannula, and by increased PCR cycles. Systemic blood was sampled by needle and pre-aspirated cannula in 101 healthy individuals with no cancer history. After erythrocyte removal, samples were subjected to RT-PCR using specific primers for CEA, with 29 or 35 RT-PCR cycles. There was a significant difference between the number of subjects whose samples were negative when collected via needle venesection and positive when collected via pre-aspirated cannula, compared with positive by needle venesection and negative by pre-aspirated cannula for both 29 (P=0.016) and 35 (P=0.011) RT-PCR cycles. Venesection technique (P=0.01) and number of cycles (P=0.003) were significant predictors of a positive result. Positive results in healthy subjects were reduced to less than 3% when an aspirated cannula was used for venesection and >29 PCR cycles were avoided.

Objective To detect the expression of CEA mRNA in patients with non-small cell lung cancer (NSCLC) and to investigate it’s 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.

PCR enzyme linked immune sorbent assay (ELISA) was developed for detection of RT-PCR-amplified cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA. The non-radioactive hybridization was performed in a streptavidin-coated microwell with digoxigenin-labeled PCR products and with biotin-labeled capture probe. PCR ELISA was proved to be expedient, simple, sensitive and safe for identification of CK19-, CEA-RT-PCR products. These results were proven by sequencing.

Objective: It has been suggested that the molecular identification of cancer cells in the circulation may be useful in predicting the presence of micrometastasis in several cancer types. The aim of the present study was therefore to assess the feasibility of CEA mRNA identification in blood for diagnosing and staging colorectal, gastric and pancreatic cancer. Methods: We studied 16 control subjects, 69 patients with colorectal (CRC), 30 with gastric (GC), 27 with pancreatic cancer (PC) and 8 with benign diseases of the pancreatobiliary tree. At diagnosis CEA mRNA was identified in peripheral blood by means of a RT-PCR procedure. Results: The specificity of this test in control subjects was 94%, and its sensitivity in identifying CRC, GC and PC were 34, 37 and 41%, respectively. False-positive findings were recorded in 25% patients with benign diseases. No association was found between CEA mRNA and stage in patients with GC or PC. In CRC patients, positive CEA mRNA findings were correlated with local spread (χ2 = 14.6, p < 0.01), lymph node (χ2 = 18.95, p < 0.001) and distant metastasis (χ2 = 11.3, p < 0.001). In these cases, CEA mRNA, but not CEA, was entered in stepwise discriminant analysis to classify the presence of lymph node metastasis. Conclusions: The molecular detection of micrometastasis in the blood by means of CEA mRNA identification is feasible for colorectal, but not for gastric or pancreatic cancer staging. Further studies are needed in order to define the clinical utility of this marker also in follow-up protocols.

The detection of tumor cells in the drainage venous blood of patients with colorectal cancer was made feasible by demonstrating carcinoembryonic antigen (CEA) mRNA in the mononuclear cell component of the blood, using a nested reverse transcription-polymerase chain reaction. CEA mRNA was detected in the drainage venous blood from 11 (42%) of 26 patients, and the rate of detection increased according to the grade of vessel invasion. CEA mRNA was detected in all patients with synchronous liver metastases, even though there was no significant correlation between the presence of CEA mRNA in the drainage venous blood and the clinicopathological findings. As the presence of CEA mRNA in the drainage venous blood is an indicator of the spread of tumor cells in patients with colorectal cancer, this assay can be used to assess the possible outcome of patients with colorectal cancer, providing one more tool for the physician-oncologist to use in designing appropriate treatments.

20068 Background: Previous RT-PCR study showed that hMAM and CEA mRNA could be detected in 95% and 78% breast carcinomas, respectively. Analyzed in normal LN, these two markers were the only ones not expressed, compared to other mRNA markers which could be detected in a variable number of cases (Marchetti 2001). We evaluate the expression of CEA and hMAM mRNA in breast cancer pts in order to investigate if the identification of these transcripts could be correlated to in the LN, PB and BM. Methods: Tumor, normal gland, LN and PB were obtained from 49 breast cancer pts who have been referred to surgery during the period of Jun/2000 to Jan/2004.BM could be obtained from 21 of these cases. Before the manipulation of the tumor, 5 mL of PB was collected and 5 mL of BM was aspirated from the iliac crest. First level axillary LN were sectioned in two. One of the halves was immediately frozen in liquid nitrogen. CEA and hMAM transcripts have been analysed by RT-PCR. Results: Tumors were positive for CEA in 38 (77.6%) cases and LN were positive in 15 cases (36.7%). 6/49 pts expressed CEA in normal breast (12.2%). Five of these, tumor also expressed CEA. Two pts (4.1%) showed CEA expression in PB and in BM. One patient expressed CEA in BM but not in PB. All pts that showed expression in PB, BM and LN, also showed transcript of CEA in tumor. hMAM transcripts were found in 39 (79.6%) normal gland tissue and in 41 of tumor samples (85.7%). LN were positive to hMAM in 12 cases (24.5%). 5 of these (10.2%) were also positive in PB and BM samples. The pts showing hMAM expression in PB, BM and LN also showed its expression in neoplasic tissue. Analyzing both markers, two pts showed the expression of CEA and hMAM in the PB and BM samples. In 21 of the 49 cases the comparison of hMAM and CEA was possible to be performed in all tissues. CEA and hMAM expression associated to larger number of LN affected. Conclusions: CEA and hMAM transcripts can be detected in majority of breast cancer tissue. Specificity of hMAM seems to be higher than CEA, which can be detected in more than 10% of non-tumor breast tissue. Synchronicity of CEA or hMAM expression in the tumor and in extra-mammary sites was high, suggesting that these markers may provide an interesting strategy for follow-up micrometastatic disease. No significant financial relationships to disclose.

4200 Background: We had previously reported that peritoneal micrometastasis determined by CEA mRNA level in peritoneal washings using real-time quantitative RT-PCR (RQ-PCR) is a good predictor for ...