CEA Expression Research Articles

Establishment and its biological characteristics of patient-derived lung cancer xenograft models

背景与目的为了更好地研究肺癌的治疗方法，建立起可靠的动物评价模型迫在眉睫。本研究的目的是研究建立肺癌原代组织块小鼠移植瘤模型的成熟方法，观察移植瘤的肿瘤生物学特性，在建模方法及基本特征等方面来证明该肿瘤模型的合理性和科学性，以期为肿瘤研究提供更有效的实验动物模型。方法取人新鲜完整肺癌组织块移植于NOD/SCID小鼠右侧前肢肩背部皮下，或经皮肺穿刺取得肿瘤小块移植于BALB/c裸小鼠肾包膜下。待皮下肿瘤增大，将其切下传代于裸小鼠右侧前肢肩背部皮下。观察移植瘤生物学特性，并取肿瘤行常规病理切片及CEA、细胞角蛋白、Ki67免疫组化检测，将原代肿瘤和移植瘤进行EGFR 18-21外显子和K-Ras 12，59外显子基因检测，采用流式细胞仪检测移植瘤细胞的细胞周期。结果本研究进行了11例肺癌组织的NOD/SCID小鼠和裸鼠建模，建成3例可多次成功传代的腺癌、小细胞肺癌和鳞癌模型。传代移植成功率高。荷瘤小鼠生长情况良好，生存期长。各代移植瘤模型的组织病理学及免疫组化表型、EGFR和K-Ras基因检测均与来源肺癌组织相一致。移植瘤细胞周期中S期延长，提示瘤细胞有增殖活性。结论本研究在国内首次利用新鲜的完整肺癌组织建成了荷肺癌NOD/SCID小鼠及裸鼠模型，并传代移植于裸鼠，建模成功率为27%。移植瘤较好地保留了人原发肺癌的恶性特征及组织病理学、生物学特性，是一种接近人体的肺癌模型，可为肺癌研究提供良好的实验平台。

Marker Expression in Circulating Cancer Cells of Pancreatic Cancer Patients

By the time patients are diagnosed with pancreatic cancer, circulating cancer cells probably exist. Therefore, the detection of pancreatic cancer cells in the peripheral circulation could be used to diagnose early pancreatic cancer, which would otherwise not be detected by current imaging methods. The expression levels of h-TERT, CK20, CEA, and C-MET were detected in a model of circulating micrometastasis in pancreatic cancer that were enriched using immune-magnetic separation of the circulating cancer cells. The sensitivity and specificity of the measurements were evaluated. The expression of the above genes was measured in the circulating cancer cells of pancreatic cancer patients. We compared their expression rate in pancreatic cancer patients at different stages to screen for the indicator with highest sensitivity and specificity for the detection of circulating pancreatic cancer cells. Immuno-magnetic nanoparticles combined with RT-PCR enabled the detection of one tumor cell per 1×10(7) peripheral blood mononuclear cells. The positive expression rates of C-MET, h-TERT, CK20, and CEA in the pancreatic cancer group were 80% (20/25), 100% (25/25), 84% (21/25), and 80% (20/25), respectively, while in the benign disease control group the rates were 0% (0/15), 0% (0/15), 6.77% (1/15), and 0% (0/15), respectively. There was a significant difference in the positive expression rate between the two groups (P<0.05). The specificity of h-TERT, CEA, and C-MET was higher than that of CK20. The positive expression rate of the four genes was not related to gender, age, tumor size, CA 19-9, or CEA serum levels (P>0.05). However, the positive expression of C-MET, CK20, and CEA closely correlated with tumor stage (P<0.05). Immuno-magnetic nanoparticles combined with RT-PCR were specific and sensitive for the detection of circulating cancer cells. The positive expression of C-MET, h-TERT, CK20, and CEA in the circulation of pancreatic patients could be used as an indicator for circulating cancer cells. The combined detection of the four genes improved the specificity and sensitivity to 100%, which may be attributable to the use of immuno-magnetic separation and enrichment of the circulating pancreatic cancer cells. Our results suggest the clinical utility of this approach.

Clinicopathologic significance of p53, hypoxia-inducible factor 1 alfa, and vascular endothelial growth factor expression in colorectal cancer.

3609 Background: Hypoxia-inducible factor 1α (HIF-1α) and vascular endothelial growth factor (VEGF) are correlated with angiogenesis and involved in tumor growth and metastases. P53 gene mutation is well established pathways in carcinogenesis and related to angiogenesis. The aim of this study was to evaluate the prognostic significances of the expressions of p53, HIF- 1α, and VEGF in colorectal cancer. Methods: The tumor tissues of 311 patients with colorectal carcinoma that underwent potentially curative resection were investigated immunohistochemically using monoclonal antibodies against p53, HIF-1α, and VEGF. Clinical information and disease-free survival and overall survival were evaluated with respect to the expressions of p53, HIF-1α, and VEGF. Results: Median follow-up duration was 48 months, and median patient age was 60 ± 11 years (range 22-82). Positivity rates of p53, HIF-1α, and VEGF were 42.4%, 63.0%, and 56.6%, respectively. HIF-1α expression in tumor tissue was found to be significantly correlated with the expression of VEGF (p = 0.040), and depth of invasion (p = 0.019). Positive relationships were found between the expressions VEGF and vascular invasion (p = 0.037), and CEA (p = 0.005), p53 expression was related to vascular invasion (p = 0.011), and CEA (p = 0.026). Univariate analysis showed that stage (p < 0.001), vascular invasion (p < 0.001), CEA (p < 0.001), and HIF-1α expression (P = 0.033) were associated with disease free survival. Stage (p < 0.001), vascular invasion (p = 0.010), CEA (p < 0.001), histological grade (p = 0.038), and HIF-1α expression (p = 0.001) were significant variables affecting overall survival. Multivariate analysis showed that stage and CEA were independently associated with disease free survival (p = 0.006, p < 0.001, respectively), and overall survival (p = 0.001, p = 0.003, respectively). HIF- 1α was independently associated with a poor overall survival (p = 0.002), however, p53 and VEGF were not found to be related to prognosis. Conclusions: HIF-1α expression in tumor tissue is associated with angiogenesis and a poor overall survival in colorectal cancer patients. No significant financial relationships to disclose.

Abstract 4038: Identification of microRNAs involved in colorectal cancer progression

Abstract MicroRNAs (miRNAs) are negative regulators of gene expression during various basic biological processes such as development, cellular differentiation, proliferation and apoptosis. Altered expression of miRNAs is associated with development and progression of various human cancers, where they mainly regulate the translation of oncogenes and of tumor suppressor genes. In colorectal cancer (CRC), these regulators complement the Vogelstein multi-step model of pathogenesis and have the potential of becoming a novel class of tumor biomarkers and of therapeutic targets. To identify miRNAs deregulated in CRC that could be used for diagnosis and prognosis, we examined by Quantitative Real-Time PCR the global expression of 378 mature miRNAs in 40 CRCs and their paired non-tumor tissues. Non-parametric confidence intervals for each miRNA where calculated and twenty-four miRNAs whose expression is significantly altered in tumors respect to the normal tissue where identified (Bonferroni P&amp;lt;0.05). Part of them, such as miR-21, miR-31 or miR-183 has been previously reported in CRC. MiR-135b, with the highest expression in the tumors is a promising tumor related miRNA. We subsequently evaluated the association of the 24 miRNAs with different clinical characteristics of the samples such as tumor site, disease status, TNM staging, expression of CEA and CA199, and presence of alterations in the molecular markers of CRC progression (APC, TP53, KRAS and loss of 18q arm). MiRNAs differently expressed among colon and rectum or higher in late stage tumors were identified. Expression levels of miR-31 were correlated to that of CA199 and miR-18a, miR-21 and miR-31 were associated with mutations in APC gene. Gene target predictions of these miRNAs pointed mainly the pathways related to CRC progression such as Wnt/TGFb, MAPK and TP53 signaling pathways. In addition, many of these miRNAs were predicted to target genes in the Vogelstein model. This study has identified promising miRNAs to be used as biomarkers (for example in plasma) in diagnostic and prognostic settings of CRC. And, in addition it shows their direct involvement in the regulation of driving events of CRC carcinogenesis which should be explored further in biological models. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4038.

Abstract 1491: CEA interacts with TGF-β receptor and inhibits TGF-β signaling in colorectal cancers

Abstract Carcinoembryonic antigen (CEA) is a tumor marker for the clinical management of colorectal cancer (CRC). The elevated blood levels of CEA are associated with metastasis and poor prognosis in CRC. There is mounting evidence that CEA enhances the metastatic potential of cancer cells. CEA increases the ability of weakly metastatic CRC to colonize the liver and to develop spontaneous hematogeneous liver and lung metastases. CEA expression has also been related with resistance to cytotoxic chemotherapy and to anoikis, a form of apoptosis caused by detachment from cell matrix. Yet the mechanism of CEA mediated metastasis is only partially understood. The TGF-β (transforming growth factor beta) signaling pathway contributes to tumorigenesis by controlling several biological processes, including cell proliferation, differentiation, migration and apoptosis. It has been reported that TGF-β regulates CEA transcription and secretion, however, little is known about the effects of CEA on TGF-β signaling. Aims: Based on the above facts, we focused on the influence of CEA on the TGF-β signaling in both normal cells and colorectal cancer cells. Results: Our preliminary data showed that CEA directly interacted with TGF-β receptors. Overexpression of CEA blocked TGF-β induced SMAD3 phosphorylation, SMAD3 translocation to nuclear and the downregulation of c-myc transcription. Targeting CEA with anti-CEA antibody rescued TGF-β response in CRC cell lines with elevated CEA expression, thereby restoring the inhibitory effects of TGF-β on the proliferation of these cancer cells. Finally, in animal experiment, we found that CEA enhanced survival of colorectal cancer cell in both local colonization and liver metastasis. Conclusion: Since CEA is a well-characterized tumor-associated antigen that is frequently overexpressed in tumors, specific antibodies targeting CEA have been developed as a novel therapeutic approach for treatment of tumors expressing CEA on their surface. Based on our study, it may be helpful to combine CEA antibody and TGF-β to inhibit cancer cell proliferation and metastasis in some cases. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1491.

Abstract 849: Detection of circulating micrometastasis in pancreatic cancer patients by immunomagnetic nanoparticles combined with RT-PCR

Abstract By the time patients are diagnosed with pancreatic cancer, circulating cancer cells probably exist. Therefore, the detection of pancreatic tumor cells in the peripheral circulation could be used to diagnose early pancreatic cancer, which would otherwise not be detected by current imaging methods. The expression levels of h-TERT, CK20, CEA, and c-met were detected in a model of circulating micrometastasis in pancreatic cancer. The sensitivity and specificity of the measurements were evaluated. The expression of the above genes was subsequently measured in the circulation of pancreatic cancer patients. We compared their expression rate in pancreatic cancer patients at different stages to screen for the indicator with highest sensitivity and specificity in the detection of circulating pancreatic cancer cells. Immunomagnetic nanoparticles combined with RT-PCR enabled the detection of one tumor cell per 1×107 peripheral blood mononuclear cells. The positive expression rates of c-met, h-TERT, CK20, and CEA in the pancreatic cancer group were 80% (20/25), 100% (25/25), 84% (21/25), and 80% (20/25), respectively, while in the benign disease control group the rates were 0% (0/15), 0% (0/15), 6.77% (1/15), and 0% (0/15), respectively. There was a significant difference in the positive expression rate between the two groups (P&amp;lt;0.05). The specificity of h-TERT, CEA, and c-met was higher than that of CK20. The positive expression rate of the four genes was not related to gender, age, tumor size, CA-199 or CEA serum levels (P&amp;gt;0.05). However, the positive expression of c-met, CK20, and CE closely correlated with tumor stage (P&amp;lt;0.05). Immunomagnetic nanoparticles combined with RT-PCR were specific and sensitive for the detection of micrometastasis. The positive expression of c-met, h-TERT, CK20, and CEA in the circulation of pancreatic patients could be used as an indicator for circulating cancer cells. The detection rate of circulating cancer cells increased to 100% by the combined measurement of the four genes, suggesting the clinical utility of this approach. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 849.

Abstract 2240: Clinical significance of CEA, PRL3 and EZH2 transcripts expressed in lymph nodes of colorectal cancer patients

Abstract Objective: The role of micrometastases detected immunologically and molecularly in lymph nodes of colorectal cancer patients remains elusive. We investigated the prognostic significance of CEA, PRL3 and EZH2 expressed in lymph nodes of 42 pathologically node-negative (pN0) colorectal patients and 13 stage III/IV patients as controls. Seventeen pN0 patients (40%) received adjuvant therapy after curative surgery. Experimental design: CEA, PRL3 and EZH2 transcripts were amplified by reverse transcription PCR in nearly 600 paraffin embedded lymph nodes. On average 9.7 ± 5.7 nodes were harvested per case. After 55 months median follow-up, gene expression was correlated with recurrence-free (RFS), overall (OS) and disease-free survival (DFS), using Kaplan-Meier analysis and log-rank test for comparison. Results: The proportion of cases demonstrating CEA+, PRL3+ and EZH2+lymph node expression increased from stage I to III/IV and reached 71%, 76% and 26% respectively in the pN0 group. EZH2 had no significant prognostic value among pN0 patients. Disease progressed more rapidly (50% DFS at 30 versus 53 months) in EZH2+ than EZH2- node-positive patients. CEA+ was associated with ∼18% lower RFS/OS, and 8% lower DFS than CEA- among therapeutically-untreated patients, those with colon and with stage II tumors. pN0 patients with PRL3+ nodes had better outcomes than PRL3- cases (DFS, P= 0.059), namely in the colorectal (OS, P= 0.056; DFS, P= 0.12), colon (DFS, P= 0.056) and stage II tumor in the non-adjuvant settings (DFS, P= 0.0047). Conclusions: Lymph node expression of the PRL3 phosphatase gene predicted more favorable outcomes for pN0 colorectal cancer patients. The role of CEA as an adverse predictor of survival was confirmed albeit less significant than previously reported. Neither CEA nor PRL3 nodal expression was predictive of rectal cancer recurrence. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2240.

Prognostic analysis of surgical resection for patients with liver metastases from colorectal carcinoma

Objective To assess the indication and efficacy of surgical treatment for patients with hepatic metastases from colorectal carcinoma. Methods Clinical data of 75 patients with hepatic metastases from colorectal carcinoma undergoing surgery from January 2000 to December 2007 were analyzed retrospectively. Results Among 75 patients,62 patients were found to have synchronous hepatic metastasis and 13 patients were found to have metachronous hepatic metastases. 70 patients underwent resection of hepatic metastases and complications occurred in 10 patients. The 1,3 and 5-year survival rates were 74. 67%, 58.67% and 28.00%respatectomy. Univariate analysis revealed that solitary metastasis was a significant factor for favorable prognosis after surgery . The expression of preoperative CEA, Dukes stage, the number of hepatic metastases, status of lymph node metastasis and histological differentiation of the primary colorectal carcinoma were all significant factors for prognosis after surgery. Conclusion The hepatic metastases from colorectal cancer should be treated by a surgical approach. T earlier stage of clinical pathology, higher differentiation extent, metastases less than 2, and low-level expression of preoperative CEA predict a better survival. Key words: Colorectal carcinoma; Liver metastasis; Surgical treatment; Prognosis

Noninvasive Imaging and Radiovirotherapy of Prostate Cancer Using an Oncolytic Measles Virus Expressing the Sodium Iodide Symporter

Prostate cancer cells overexpress the measles virus (MV) receptor CD46. Herein, we evaluated the antitumor activity of an oncolytic derivative of the MV Edmonston (MV-Edm) vaccine strain engineered to express the human sodium iodide symporter (NIS; MV-NIS virus). MV-NIS showed significant cytopathic effect (CPE) against prostate cancer cell lines in vitro. Infected cells effectively concentrated radioiodide isotopes as measured in vitro by Iodide-125 ((125)I) uptake assays. Virus localization and spread in vivo could be effectively followed by imaging of (123)I uptake. In vivo administration of MV-NIS either locally or systemically (total dose of 9 x 10(6) TCID(50)) resulted in significant tumor regression (P < 0.05) and prolongation of survival (P < 0.01). Administration of (131)I further enhanced the antitumor effect of MV-NIS virotherapy (P < 0.05). In conclusion, MV-NIS is an oncolytic vector with significant antitumor activity against prostate cancer, which can be further enhanced by (131)I administration. The NIS transgene allows viral localization and monitoring by noninvasive imaging which can facilitate dose optimization in a clinical setting.

Detection of tumor cells in peripheral blood of patients with gastric cancer using mRNA of MAGE genes as markers

To develop a sensitive and specific RT-PCR assay using the mRNA of MAGE-1 and MAGE-3 genes as specific tumor markers for the detection of the tumor cells in the peripheral blood of patients with gastric cancer. Peripheral blood was obtained from 40 patients with gastric cancer and from 20 healthy volunteers. The mRNA of MAGE-1 and MAGE-3 genes in the peripheral blood mononuclear cells (PBMC) was detected by RT-PCR. The expressions of MAGE-1 and MAGE-3 mRNA in the tumor tissues of these gastric cancer patients were also detected by RT-PCR. Meanwhile,CEA expression by nested RT-PCR in PBMC of 40 gastric cancer patients was also detected. Of 40 gastric cancer patients, MAGE-1 and MAGE-3 mRNA were positive in 47.5% (19/40) and 25% (10/40) of PBMC respectively, and in 62.5% (25/40) and 30% (12/40) of gastric cancer tissues respectively. As a whole, in the PBMC of 40 gastric cancer patients, 25 (62.5%) samples were found to express at least one type of MAGE mRNA. In the patients whose tumors did not express MAGE-1 and/or MAGE-3 genes, the corresponding MAGE mRNA was also undetected in their PBMC. There was no expression of MAGE-1 or MAGE-3 gene in the PBMC from the 20 healthy donors. The positive rate of MAGE mRNA in PBMC was closely correlated with the tumor stage and lymph node metastasis (P <0.05). Positive rate of CEA gene expression was 32.5% (13/40) in the PBMC of 40 gastric cancer patients, 29 (72.5%)samples were detected to express at least one type of MAGE gene and CEA gene mRNA. MAGE-1, MAGE-3 and CEA mRNA are specifically detected with high percentage in the PBMC of gastric cancer patients by RT-PCR. They could be used as specific tumor markers for the detection of the circulating gastric cancer cells, and the detection results may be helpful to evaluate the prognosis of gastric cancer patients.

A new way for isolation and cultivation of sweat gland ductal cells from human split-thickness skin in vitro

To explore a new method of isolation and culture of eccrine sweat gland ductal cells from human split-thickness skin graft in vitro. Human split-thickness skin graft which was presented by volunteer (n = 10) was digested with type II collagenase, and then sweat gland duct were isolated from the split-thickness skin graft, primary cultures were incubated at 37 degrees C in humidified atmosphere of 5% CO2, 95% O2. The cultured eccrine sweat gland ductal cells were identified by analysis CEA, CK8, CK18, CK19 antigens expression with flow cytometry, RT-PCR and Western Blot, and by detecting the electrophysiology with whole cell patch clamp technology. The isolated eccrine sweat gland ductal cells could grow by adhering to the wall, proliferate in vitro after 48 h of adhering to the wall, and confluens after 2 - 4 weeks of adhering to the wall. The FACs analysis showed the expression of CEA was (90.26 +/- 1.12)%, (89.70 +/- 1.43)%, and CK8 was (94.41 +/- 1.84)%, (93.65 +/- 1.63)% in primary cultured sweat gland ductal cells and primary cultured eccrine sweat gland cells, respectively, and there is no significant difference between the two groups (P > 0.05). Immunocytochemistry staining showed CEA, CK8, CK18, CK19 was positive in sweat gland duct cells, RT-PCR revealed that CEA, CK8, CK18 and CK19 gene expression in sweat gland ductal cells, and Western Blot analysis showed the expression of CEA brand, CK8 brand, CK18 brand, and CK19 brand in sweat gland ductal cells, patch clamp indicated that this cells has distinct amiloride sensitive Na(+) channels. The cultured human eccrine sweat gland duct cells in vitro display the markers and biological characteristics of sweat gland epithelial lineage, and this method of digest the split-thickness skin graft to get the sweat gland duct cells is better than classical dissect sweat gland under dissect microscope.

Clinical Significance of CK20, CK19, CEA mRNAs in Peripheral Blood from Lung Cancer Patients.

The tumor cells may disseminate into circulation even in early stages. However, the numbers of such cells are so small that they cannot be detected by conventional diagnostic methods. Developments in molecular technology have made it possible to detect small numbers of tumor cells in peripheral blood. In this study, 83 preoperative peripheral blood samples from patients with lung cancer were analyzed for the determination of CEA, CK19, and CK20 mRNA expression in peripheral blood, and its clinical significance was evaluated. Nested reverse transcriptasepolymerase chain reaction (nested RT-PCR) was used to analyze CEA, CK19, and CK20 mRNA expression in peripheral blood. Fresh tumor tissues from patients with esophageal cancer (n=15) were used as a positive control, and blood samples from 15 healthy volunteers as a negative control. Among 83 blood samples from patients with lung cancer, the expression of CK20, CK19 and CEA mRNA was 41.0%, 36.1% and 48.2%, respectively. 73.5% were positive for at least one markers for these three genes, and were significantly correlated with the metastasis of the cancer (P<0.05), but had no correlation with age and sex. In primary esophageal tumors, 15 tumor tissues were all positive for CEA, CK19, and CK20 mRNA. 2 of 15 blood samples from healthy donors were positive for CK19 and CEA mRNA, and 1 of 15 blood sample positive for CK20 mRNA. The three markers of CK20, CK19 and CEA mRNA could be the target genes in detecting circulating tumor cells in peripheral blood from patients with lung cancer. The combined detection of CK20, CK19 and CEA mRNA by RT-PCR assay may contribute to the PCR positivity and sensitivity, and CK20, CK19, CEA mRNA expression increase the specificity.

W1509 CEA and SCC-Antigen mRNA Expression in Preoperative Peripheral Blood Is a Significant Prognostic Factor in Patients with Esophageal Cancer

S A T A b st ra ct s interval [CI] 1.038-5.258, P=0.04) and age >40 years (OR 1.939, SE 0.472, CI 1.203-3.126, P=0.007) to be associated with mortality. Male gender, primary operative treatment and the coincidence of multiple predisposing factors were not related with mortality. Neoplastic disorders were associated with an increased ASR-related mortality (OR 2.634, SE 0.541, CI 1.762-3.938, P=0.008) compared to the other etiological groups. Patients with an atraumaticidiopathic splenic rupture (normal spleen, group f) showed a decreased ASR-related mortality (OR 0.111, SE 0.112, CI 0.015-0.807, P=0.03). CONCLUSIONS: Most ASR are atraumaticpathologic splenic ruptures. Splenomegaly and advanced age are negative predictive factors for survival. Neoplastic disorders are associated with an increased rupture-related mortality.

Expression and clinical significance of CEA-mRNA, CK20-mRNA in peripheral blood of esophageal cancer patients

Objective To establish reverse transcriptase-polymerase chain reaction(RT-PCR) with primers specific for CEA gene and CK20 gene to detect circulating tumor cells in peripheral blood of esophageal cancer pa-tients,and try to find the relationship between the mRNA expression and micrometastasis. Methods The expressions of CEA,CK20 were analyzed by RT-PCR in 53 cases of esophageal tumor tissue and in peripheral blood,compared with 10 patients with benign esophageal disease and 20 healthy volunteers. Results The expressions of CEA-mRNA, CK20-mRNA were 96.23% (51/53), 100% ( 53/53 ) in 53 esophageal tumor tissue and were 52.83% (28/53), 49.06% (26/53) in peripheral blood of 53 esophageal cancer patients. In control group,there was only one expression of CEA-mRNA in peripheral blood of 10 patients with benign esophageal disease,as well as in 20 healthy volunteers. There was no expression of CK20-mRNA in peripheral blood of 10 patients with benign esophageal disease and 20 healthy volunteers. Conclusion CEA-mRNA, CK20-mRNA might be specific and sensitive markers to detect circu-lating tumor cells in peripheral blood and their expression was closely related to TNM stages of the esophageal cancer patients. Key words: Esophageal neoplasms; Neoplasm metastasis; Carcinoembryonic antigen; Keratin-20

Clinicopathological and immunohistochemical features of colorectal sessile serrated adenoma

To investigate the clinicopathological characteristics and expression status of Ki67, p53, CEA, CDX, CK7 in colorectal sessile serrated adenoma (SSA). The clinicopathological data of 11 cases of SSA, 51 cases of hyperplastic polyp (HP) and one case with mixed HP/SSA were reviewed and analyzed retrospectively. The expression of Ki67, p53, CEA, CDX and CK7 were detected by immunohistochemistry. The major histological features in SSA were architectural abnormality in crypts, dilatation of serrated crypt bases like an inverted "T" or "L" shape adjacent to muscularis mucosa. Atypical cells containing round to oval nuclei and nucleoli were also observed. The immunohistochemical staining showed that the expression of p53 increased gradually from HP to TA: 11.8% in HP, 20.0% in SSA, 41.2% in VTA and 75.0% in TA, with a significant difference among the groups (chi(2) = 17.996, P = 0.000). However, no significant difference in the expression of CDX and CK7 was observed between HP and SSA. Of the 10 SSA cases, positive expression of Ki67 was found in cells located in the base or middle part of crypt in 6 cases, positive cells index was 26% - 50% in 5 cases, and > 50% in 3. Compared with the expression of Ki67 in the HP, VTA and VA, SSA showed a significant difference in both the positive cell number and in the positive regions. (positive number: chi(2) = 34.601, P = 0.000; positive regions: chi(2) = 63.077, P = 0.000). Morphological diagnosis of SSA was mainly based on crypt architectural and cellular abnormalities, and the crypt architectural abnormality may be more important than cellular features. Detection of p53 and Ki67 expression may be helpful in differential diagnosis and understanding the nature of SSA.

Rare Tumor of the Tracheobronchial Tree: Solitary Squamous Papilloma

A 64-year-old woman presented with an intractable cough and purulent sputum, and a chest CT revealed a mass within the left lower lobar bronchial orifice. A flexible bronchoscopy demonstrated a red, glistening, nodular single lesion, obstructing the lumen of the left lower lobe of the bronchus, and biopsy of the specimen showed squamous papilloma. Surgical resection was selected for the definite treatment, and a simple left lower lobectomy was performed. Histopathological examination revealed a 1.2 x 1.0 cm sized bronchial obstructing mass and the diagnosis of squamous papilloma was confirmed with low p53, Ki-67 (less than 5 %) and focal weak positive CEA expression. The patient had an uncomplicated postoperative course and remains asymptomatic 3 months afterwards.

Expression and clinical significance of ADAM8 and CEA in serum of patients with non-small cell lung cancer.

Lung cancer is one of the most common cancers in the world, whose mortality is at the first place in all malignant tumors. This study is to investigate the diagnostic value of ADAM8 and CEA in serum of non-small cell lung cancer patients. The serum levels of ADAM8 were assayed by ELISA in 62 NSCLC patients, 27 benign palmonary lesions and 32 healthy subjects as control. The serum level of CEA were assayed by radioimmunoassay. The serum level of ADAM8 and CEA in group of NSCLC were both remarkably higher than those in lung benign lesions and normal controls (P <0.01), without significant difference between benign palmonary lesions and normal controls (P >0.05). There was no significant difference between adenocarcinoma and squamous cell carcinoma in serum ADAM8 level (P >0.05). The serum level of CEA in adenocarcinoma were significantly higher than that in squamous cell carcinoma (P <0.05). The serum level of ADAM8 and CEA in NSCLC with stages III-IV were both remarkably higher than those with stages I-II (P <0.01). The level of ADAM8 and CEA in NSCLC with lymph node metastasis positive were both higher than those of negative (P <0.01). The diagnostic sensitivity of ADAM8 and CEA for NSCLC was 77.4% and 71.0% respectively, and the specificity was 90.6% and 84.4% respectively. The combined detection of ADAM8 and CEA could improve the sensitivity to 91.9%, but the specificity decreased to 75.0%. The overexpression of ADAM8 in serum of NSCLC patients indicates that ADAM8 is related to the development of NSCLC; Combined detection of ADAM8 and CEA is helpful for the diagnosis of NSCLC.

Relationship between expression of CEA, E-cadherin and liver metastasis in colorectal cancer

Objective To investigate the expression of E-cadherin and CEA in serum in colorectal carcinoma and their relationship with liver metastasis.

Balancing between Antitumor Efficacy and Autoimmune Pathology in T-Cell–Mediated Targeting of Carcinoembryonic Antigen

Carcinoembryonic antigen (CEA) is intensively studied as a potential target for immunotherapy of colorectal cancers. Although overexpressed by tumors, CEA is also expressed in normal tissues, raising questions about the feasibility and safety of CEA-targeted immunotherapy. We investigated these issues in transgenic mice in which the expression of human CEA in normal tissues closely resembles that in man. Our data show that the T-cell response against CEA in these mice is blunted by both thymic and peripheral tolerance. Consequently, effective tumor targeting is only achieved by adoptive transfer of T cells from nontolerant donors in combination with interventions that eliminate peripheral immune regulatory mechanisms. However, such treatments can result in severe intestinal autoimmune pathology associated with weight loss and mortality. Interestingly, preconditioning of recipient mice by depletion of T-regulatory cells results in immune-mediated tumor control in the absence of toxicity. In this setting, CEA-specific T-cell responses are lower than those induced by toxic regimens and accompanied by additional T-cell responses against non-self antigen. These findings illustrate the importance of testing adoptive immunotherapies targeting self antigens such as CEA in preclinical in vivo models and show that the choice of immune intervention regimen critically determines the balance between therapeutic efficacy and toxicity.

Detection of colorectal cancer cells from feces using quantitative real‐time RT‐PCR for colorectal cancer diagnosis

Early detection of colorectal cancer (CRC) is desired for reducing its mortality rate. Recently, the feasibility of a new method for isolating colonocytes from feces was demonstrated, followed by direct sequencing analysis for detecting colorectal cancer. In the present study, gene expression analysis was conducted using quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR). One hundred and sixty-six patients with CRC and 134 healthy volunteers were enrolled. Messenger RNA expressions of CEA, MMP7, MYBL2, PTGS2 and TP53 in the colonocytes isolated from feces were analyzed by quantitative real-time RT-PCR. Beta-2-microglobulin, used for internal control, could not be detected in approximately 25% each of the CRC patients (39/166) and healthy volunteers (33/134). CEA expression did not differ significantly between CRC patients and healthy volunteers (P = 0.21). MMP7, MYBL2, PTGS2 and TP53 gene expressions were significantly higher in CRC patients than in healthy volunteers (P < 0.001). The overall sensitivity and specificity using these gene expressions were 58.3% (74/127, 95% CI; 49.2-67.0) and 88.1% (89/101, 95% CI; 80.2-93.7), respectively. The sensitivity was dependent on the tumor location (P = 0.01) and tumor size (P = 0.02), but not the tumor depth (P = 0.06) or cancer stage (P = 0.37). Gene expression analysis of colonocytes isolated from feces may be a useful method for CRC screening, if the number of isolated colonocytes is sufficiently high for analysis by quantitative real-time PCR. Therefore, improvement of the colonocyte retrieval system from feces may be necessary for the technique to be developed for clinical use.