CEA Expression Research Articles

Primary cutaneous keratinocytic primordial carcinoma with triple differentiations into basal cell carcinoma, sqaumous cell carcinoma, and porocarcinoma

The author herein reports a hitherto undescribed entity of cutaneous primordial carcinoma with triple differentiation into basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and porocarcinoma (PC) in a 66-year-old man. The patient was treated by tumor excision with wide margins. Pathologically, the margins were negative for tumor cells. The patient is now healthy 1 year after the excision. The tumor measured 3 cm × 2 cm × 2 cm and arose from the epidermis. The percentage of the occupying area was 60% in BCC, 20% in SCC, and 20% in PC. Histologically, BCC was characterized by basalioma cells, melanin pigmentation, peripheral palisading, and cleft formation, SCC by ample acidophilic cytoplasm, individual keratinization and cancer pearls, and PC by poroma cells and tubular structures containing cuticles. Cystic changes in BCC and SCC, keratotic nature of BCC, and acatholytic features of SCC were also seen in places. There were gradual merges among the three components. Immunohistochemically, BCC component was characterized by strong and diffuse expression of KIT, PDGFRA and p63, weak expressions of cytokeratin (CK) AE1/3, CK CAM5.2, CK34BE12, and negative expressions of CK18, CK19, CEA and CA19-9. SCC component was characterized by strongly positive expressions of p63, CEA, CK34BE12, CK7 and CK8, and negative expressions of CK18, CK19, and CA19-9. PC was characterized by strongly positive expressions of KIT, PDGFRA, CK34BE12, CK7, CK8, CK18, CK19, CEA, S100 protein, CA19-9, MUC1, and negative expression of p63. The present case revealed that there are primary cutaneous keratinocytic primordial carcinomas with triple differentiations into BCC, SCC, and PC. The present case is the first example of such tumors. It is thought that KIT and PDFRA-positive stem cells or cancer stem cells in the BCC component give rise to the SCC and PC components in the present case.

Abstract 4272: Organoids and patient-derived tumor xenograft of pancreatic adenocarcinoma share morphological and genetic features with the primary tumor

Abstract Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer death, with a 5-year overall survival rate <7%. Several active systemic therapies are now available for PDAC. Personalizing therapy may be improved with the development of realistic tumor models from a patient's explanted tumoral tissue. Human pancreatic tumor patient-derived xenografts (PDX) implanted into immunodeficient mice and tumor organoids grown in vitro 3D culture are two promising models. However, it is uncertain if these models are histopathologically and genetically similar to the primary PDAC. Histopathological comparison of sections from PDX tumor, organoids, and primary PDAC from a 63-year-old female patient were performed on paraffin embedded tissue. H&E and immunohistochemical staining for cytokeratins (CK7, CK20), p53, Claudin-4, and CEA were performed. DNA and mRNA sequencing was performed. Both PDX and organoids exhibited histopathological features remarkably consistent with the original patient tumor including the histologic grade (moderately differentiated), cytological appearance (irregular nuclear membranes, open chromatin and prominent nucleoli), mitotic activity (5 -7/10 HPF), and immunohistochemical profile. The PDX and organoids demonstrated diffuse moderate-to-strong positivity for CK7, CEA, p53, and Claudin-4 and focal weak positivity for CK20 - all similar to the primary tumor. The immunohistochemical staining pattern was consistent with mRNA sequencing of the primary tumor which showed that CEA, Claudin-4 and p53 expression were ∼960-fold, ∼27-fold and ∼3 fold higher respectively (vs. benign pancreatic tissue). DNA sequencing revealed somatic mutations in KRAS and TP53 genes seen in >90% and ∼70% of PDACs respectively, and a few rare somatic mutations, such as a sodium leak channel (NALCN) mutation, seen in ∼3% of PDAC. Both PDX and organoid models of PDAC maintain key histological features, immunohistochemical profile and basic gene expression pattern akin to the primary tumor. These findings suggest that PDXs and organoids have the potential to serve as reliable pathophysiological models for optimizing individual therapy for patients with PDAC. Citation Format: Isabel Romero Calvo, Ashwin Akki, Andrey Ugolkov, Mary M. Buschmann, Samantha M. Sparrow, Teresa Barry, Margaret Eber, Tongjun Gu, Shuang Qin Zhang, Hedy Kindler, William Dale, Kevin Roggin, Andrew P. Mazar, Kevin P. White, Christopher R. Weber. Organoids and patient-derived tumor xenograft of pancreatic adenocarcinoma share morphological and genetic features with the primary tumor. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4272.

Analysis of circulating tumor cells in colorectal cancer liver metastasis patients before and after cryosurgery

ABSTRACTIn this study, we determined the number of peripheral blood circulating tumor cells (CTCs) pre- and post-cryosurgery in patients with colorectal cancer liver metastasis as a reference for understanding the relevance of any changes to the efficacy of cryosurgery. CTC numbers and CTC-related gene expression were measured in the peripheral blood of 55 patients with colorectal liver metastasis at 1 day before and 7 and 30 d after cryoablation using magnetic activated cell sorting (MACS) and fluorescence activated cell sorting (FACS) combined with real-time quantitative PCR (RT-qPCR). The number of CTCs decreased significantly with postoperative time (P < 0.01). Delta cycle threshold values for the CTC-related genes CEA, Ep-CAM, CK18 and CK19 increased significantly after cryoablation. Furthermore, the expression of CEA, Ep-CAM, CK18 and CK19 decreased significantly with time after cryoablation (P < 0.01). RT-qPCR and FACS combined with MACS has significant diagnostic and prognostic value for evaluating the efficacy of cryosurgery in patients with advanced colorectal cancer.

Exosomes from human colorectal cancer induce a tumor-like behavior in colonic mesenchymal stromal cells

BackgroundCancer cells, including colorectal cancer ones (CRC), release high amounts of nanovesicles (exosomes), delivering biochemical messages for paracrine or systemic crosstalk. Mesenchymal stromal cells (MSCs) have been shown to play contradicting roles in tumor progression.ResultsCRC exosomes induce in cMSCs: i) atypical morphology, higher proliferation, migration and invasion; ii) formation of spheroids; iii) an acidic extracellular environment associated with iv) a plasma membrane redistribution of vacuolar H+-ATPase and increased expression of CEA. Colon cancer derived MSCs, which were isolated from tumor masses, produce umbilicated spheroids, a future frequently observed in the inner core of rapidly growing tumors and recapitulate the changes observed in normal colonic MSCs exposed to CRC exosomes.Materials and MethodsTissue specific colonic (c)MSCs were exposed to primary or metastatic CRC exosomes and analysed by light and electron microscopy, proliferation in 2D and 3D cultures, migration and invasion assays, Western blot and confocal microscopy for vacuolar H+-ATPase expression.ConclusionsCRC exosomes are able to induce morphological and functional changes in colonic MSCs, which may favour tumor growth and its malignant progression. Our results suggest that exosomes are actively involved in cancer progression and that inhibiting tumor exosome release may represent a way to interfere with cancer.

A Novel Carcinoembryonic Antigen T-Cell Bispecific Antibody (CEA TCB) for the Treatment of Solid Tumors.

CEA TCB is a novel IgG-based T-cell bispecific (TCB) antibody for the treatment of CEA-expressing solid tumors currently in phase I clinical trials (NCT02324257). Its format incorporates bivalent binding to CEA, a head-to-tail fusion of CEA- and CD3e-binding Fab domains and an engineered Fc region with completely abolished binding to FcγRs and C1q. The study provides novel mechanistic insights into the activity and mode of action of CEA TCB. CEA TCB activity was characterized on 110 cell lines in vitro and in xenograft tumor models in vivo using NOG mice engrafted with human peripheral blood mononuclear cells. Simultaneous binding of CEA TCB to tumor and T cells leads to formation of immunologic synapses, T-cell activation, secretion of cytotoxic granules, and tumor cell lysis. CEA TCB activity strongly correlates with CEA expression, with higher potency observed in highly CEA-expressing tumor cells and a threshold of approximately 10,000 CEA-binding sites/cell, which allows distinguishing between high- and low-CEA-expressing tumor and primary epithelial cells, respectively. Genetic factors do not affect CEA TCB activity confirming that CEA expression level is the strongest predictor of CEA TCB activity. In vivo, CEA TCB induces regression of CEA-expressing xenograft tumors with variable amounts of immune cell infiltrate, leads to increased frequency of activated T cells, and converts PD-L1 negative into PD-L1-positive tumors. CEA TCB is a novel generation TCB displaying potent antitumor activity; it is efficacious in poorly infiltrated tumors where it increases T-cell infiltration and generates a highly inflamed tumor microenvironment. Clin Cancer Res; 22(13); 3286-97. ©2016 AACR.

CEA TCB: A novel head-to-tail 2:1 T cell bispecific antibody for treatment of CEA-positive solid tumors

ABSTRACTCarcinoembryonic antigen T cell bispecific antibody (CEA TCB) is a bispecific antibody used to recognize CEA and CD3e via a novel molecular format (2:1) that induces T cell-mediated killing of CEA over-expressing tumors while sparing primary cells with low CEA expression. CEA TCB treatment inhibits tumor growth and generates a highly inflamed tumor microenvironment.

Flow cytometry as the new ‘gold standard’ for detection of free tumour cells in abdominal lavage fluid in gastric cancer patients: A comparative study of molecular and conventional methods

AimThe ideal method for detection of free tumour cells in abdominal lavage fluid should be rapid, reliable and widely available. Flow cytometry potentially covers these properties, but there are only a few studies directly comparing flow cytometry for detecting gastric cancer cells to other methods. Therefore, we compared free tumour‐cell detection in abdominal lavage fluids using cytology with immunocytochemistry, RT‐qPCR and flow cytometry.MethodsPeritoneal lavage fluid samples were collected from 10 patients. Detection of free tumour cells was performed using cytological and immunocytochemical analysis, and using RT‐qPCR and flow cytometry. Tumour cells were detected according to CEA and CK20 mRNA expression levels using RT‐qPCR, and with the epithelial markers EpCAM/CD326 and CEA using flow cytometry.ResultsThe sensitivity and specificity of cytology were 40% and 100%, respectively, while the RT‐qPCR had a sensitivity of 80% and specificity of 100%. Flow cytometry showed no false‐negative results and only one false‐positive result, giving a sensitivity and specificity of 100% and 80%, respectively. The RT‐qPCR and flow cytometry could detect free tumour cells in each of the three false‐negative cases with cytology. In the samples with detected tumour cells, tumour cell clusters were observed with imaging flow cytometry, providing additional morphological confirmation.ConclusionThe combination of quantitative and qualitative data from flow cytometry and the images of tumour cells in borderline cases provide a rapid, reliable and reproducible method for detection of free tumour cells in abdominal lavage fluid, which is necessary for intraoperative selection of patients for intraperitoneal chemotherapy.

Expression of CEA, CA19-9, CA125, and EpCAM in pseudomyxoma peritonei

Pseudomyxoma peritonei is a fatal clinical syndrome with mucinous tumor cells disseminated into peritoneal cavity and secreting abundant mucinous ascites. The serum tumor markers CEA, CA19-9, and CA125 are used to monitor pseudomyxoma peritonei remission, but their expression at tissue level has not been well characterized. Herein, we analyzed expression of these proteins and the adenocarcinoma marker EpCAM in 92 appendix-derived pseudomyxoma peritonei tumors by immunohistochemistry. All tumors were found to ubiquitously express CEA and EpCAM. In the majority of the tumors (94.6%), CEA showed polarized immunostaining, but in 5 aggressive high-grade tumors containing numerous signet ring cells, a nonpolarized staining was detected. We found preoperative CEA serum values to correlate with peritoneal cancer index. However, the serum values of the advanced cases with nonpolarized staining pattern were normal, and the patients died within 5 years after diagnosis. Thus, serum CEA measurements did not reflect aggressiveness of these tumors. CA19-9 showed strong immunopositivity in most of the tumors (91.3%), and mutated enzyme FUT3 was demonstrated from the cases showing negative or weak staining. CA125 was infrequently expressed by tumor cells (focal staining in 6.5% of the cases), but in most of the cases (79.3%), adjacent nonneoplastic mesothelial cells showed immunopositivity. As a conclusion, CEA and EpCAM are invariably expressed by pseudomyxoma peritonei tumor cells and could be exploited to targeted therapies against this malignancy.

Combination of IL-6, IL-10, and MCP-1 with traditional serum tumor markers in lung cancer diagnosis and prognosis.

Early detection and treatment is critically important for lung cancer patients. Inflammatory mediators such as IL-6, IL-10, and MCP-1 participate in lung cancer regulation. CEA, CA125, and ProGRP are commonly used serum tumor markers for lung cancer. In this study, we assessed the sensitivity and specificity of CEA, CA125, and ProGRP when used in combination with IL-6, IL-10, and MCP in lung cancer diagnosis. Serum from three different groups (healthy controls, individuals with high risk for lung cancer, and lung cancer patients) was collected. Electrochemiluminescence was used to detect expressions of CEA, CA125, and ProGRP; ELISA was used to examine serum levels of IL-6, IL-10, and MCP-1. Specificity and sensitivity of single as well as combination markers in lung cancer diagnosis were determined. Results indicated that CEA, CA125, ProGRP, and MCP-1 were significantly up-regulated in lung cancer patients as compared to those in controls and high risk individuals. Higher IL-6 and IL-10 levels were observed in both lung cancer patients and high-risk individuals as compared to those in controls. Highest sensitivity (95.2%) in cancer diagnosis was achieved when all six markers were used. This was followed by a combination of IL-6, IL-10, CEA, CA125, and ProGRP (92.6%). The most sensitive (88.6%). Four-marker combination was composed of IL-6, CEA, CA125, and ProGRP. As the combined usage of CEA, CA125, ProGRP, IL-6, IL-10, and MCP-1 significantly improved sensitivity of lung cancer detection; this biomarker arrangement may be beneficial for early diagnosis, treatment, and prognosis of lung cancer.

Survivin and VEGF as Novel Biomarkers in Diagnosis of Endometriosis.

SummaryBackgroundThe aim of this study was to investigate the role of peripheral blood markers as additional diagnostic tools to transvaginal ultrasound (TVU) findings in the diagnosis of endometriosis.MethodsThis study included 40 patients undergoing laparoscopy for suspected endometriosis from January to December 2012. Preoperative levels of serum CA125, CA19-9, CEA and mRNA expression levels for survivin and VEGF were obtained. Real-time PCR was used to determine relative gene expression. A new diagnostic score was obtained by deploying the peripheral blood markers to the TVU findings. Statistical methods used were Chi-square, Fisher’s, Student’s t-test or the Mann – Whitney test.ResultsThere was a statistically significant difference in serum CA125, survivin and VEGF levels in patients with endometriosis and those without endometriosis (p<0.001, p=0.025 and p=0.009, respectively). False negative TVU findings were noted in 3/13 patients (23.1%) with peritoneal endometriosis without ovaries involvement. High sensitivity (93.3%), specificity (90.0%), PPV (96.6%), NPV (81.8%) and accuracy (92.5%) were obtained for a diagnostic score based on TVU and significant peripheral blood markers (CA125, survivin and VEGF).ConclusionsDetermination of serum CA125, mRNA expression levels for survivin and VEGF along with TVU can contribute to higher accuracy of the noninvasive diagnostic tools for endometriosis.

CEA Expression by Malignant Cells in an Endometrial Biopsy Not Always Endocervical Carcinoma

CEA Expression by Malignant Cells in an Endometrial Biopsy Not Always Endocervical Carcinoma

Abstract 67: Genomic and mutational profiling of human colon adenomas reveals early driver mutations and a TGF-β-CEA regulated profile

Abstract Introduction: Colorectal cancer (CRC) is the third most commonly diagnosed cancer in the world with 143,700 newly diagnosed cases in 2012. The rapid development of cancers in small, sessile adenomas could be a contributing factor that may have been overlooked. Identifying high risk patients through genomic analysis of adenomas could potentially lead to an early intervention therapy. Methods: Whole Genome Sequence (WGS) and Whole Exome Sequence (WES) analyses were performed for 4 pairs of normal controls (two from proximal colon and two from the distal colon) and colorectal adenomas (2 sessile serrated (SSA) and 2 tubulovillous adenomas (TVA) less than 1 cm in size). Transcriptome sequence analysis was performed in seven pairs of control-test matched adenomas (6 TVA and 1 SSA). Expression level of CEA and TGF-β pathway members were carried out on 30 non dysplastic adenomas and normal colon tissues by immunohistochemistry. Results: 1. Hyper-mutator profiles were observed in two of the samples (1 SSA and 1 TVA) by WGS with 1709 mutations after normalization with normal paired samples. 2. The samples showed an average mutation frequency of 0.55 mutations per 106 bases. 3. Aberrant mutational profiles was detected in seven of the eleven adenomas, with distinct mutational signatures among the samples, two with high, two intermediate and three low mutational rates. 4. Six of the eleven adenomas (1 SSA and 4 TVAs) showed alteration in the Wnt and p53 pathway. 5. Transitional single nucleotide substitutions of C:T&amp;gt;G:A in the mutational spectrum were observed in 37% of the samples. 6. Subtle localized hyper mutation (kataegis) was observed among two of the samples. 7. Five out of eleven adenomas showed mutations in the TGF-β (transforming growth factor-β) and CEA pathways members, overlapping with Wnt/p53 mutations in four adenomas. 8. Analyses of expression level of CEA and TGF-β pathway members in 30 non dysplastic tissues revealed a marked increase (over 8 fold) in CEA expression in 25% of adenoma samples which was linked to concomitant loss of TGF-β signaling. 9. Further functional studies revealed that CEA associated with TGF-β Type I receptor and disruption of TGF-β tumor suppressor signaling with activation of STAT3. Conclusions: Our studies indicate that small adenomas both TVAs and SSAs can resemble CRCs in genomic profiling and may reflect a high risk population. Genetic and mechanistic analyses reveal that disruption of CEA/TGF-β pathway in early adenomas may reflect a new and early role for these pathways in CRC. This study further supports the biomarker driven targeting of CEA/TGF-β in high risk adenomas and can be used as a prognostic marker for early detection of aggressive adenoma-CRC progression. Citation Format: Vipin K. Menon, Raju S. Gottumukkala, Jian Chen, Xiaoping Su, Nipun Mistry, Avijit Majumdar, Ji-Hyun Shin, Shulin Li, Kirti Shetty, Xifeng Wu, Brian Weston, Ethan Miller, John R. Stroehlein, Marta L. Davila, Mehnaz A. Shafi, Asif Rashid, Bhaskar V. Kallakury, Selvi Thirumurthi, John S. McMurray, Sue-Hwa Lin, Wilma Jogunoori, Lopa Mishra. Genomic and mutational profiling of human colon adenomas reveals early driver mutations and a TGF-β-CEA regulated profile. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 67. doi:10.1158/1538-7445.AM2015-67

Morphological and immunohistochemical peculiarities of placental-endometrial structures in women with benign and malignant tumors

Aim. Researchmaterials were the placenta of women operated on papillary carcinoma (encapsulated), endometrium of women with the same tumor markers detected in the placenta, this endometrium were taken during diagnosing postpartum catamnesis of the state of reproductive health, in comparison to the endometrium of women with benign uterine tumors (leiomyoma).Methods and results. Methods, that were used in the study: histological, immunohistochemical – indirect streptavidin-peroxidase method for detecting the expression of Ki67 Clon MIB-1, p53 Clon: DO-7, REA, Cytoceratina AE1 / AE3; Vimentin Clone: v9, receptors for estrogen (RE) and progesterone (RP). The study evaluated predictors of placental-endometrial disorders in women with cancer pathology: elevation of relative amount of expression of proliferative marker Ki-67 and p53 tumor marker in the placenta and endometrium, leading to violations in the structure of the process of regeneration; dischronosis in levels of expression of estrogen and progesterone receptors in the endometrium; presence of CEA expression in 25% of women of the main group in the endometrium and placenta; atypical glandular hyperplasia, endometrial polyps and micropolyps were found, which are important in the development of oncologic pathology.

The prognostic significance of pretreatment serum CEA levels in gastric cancer: a meta-analysis including 14651 patients.

BackgroundCarcinoembryonic antigen (CEA) is commonly used as a serum tumor marker in clinical practice; however, its prognostic value for gastric cancer patients remains uncertain. This meta-analysis was performed to assess the prognostic value of CEA and investigate CEA as a tumor marker.MethodsPubMed, EMBASE and other databases were searched for potentially eligible studies. Forty-one studies reporting the prognostic effect of pretreatment serum CEA expression in gastric cancer patients were selected. Data on 14651 eligible patients were retrieved for the meta-analysis. Based on the data extracted from the available literature, the hazard ratio (HR) and 95% confidence interval (CI) for an adverse prognosis were estimated for gastric cancer patients with elevated pretreatment serum levels of CEA (CEA+) relative to patients with normal pretreatment CEA levels (CEA-).ResultsThe CEA+ patients had a significantly poorer prognosis than the CEA- patients in terms of overall survival (OS: HR 1.716, 95% CI 1.594 - 1.848, P< 0.001), disease-specific survival (DSS: HR 1.940, 95% CI 1.563 - 2.408, P< 0.001), and disease-free survival (DFS: HR 2.275, 95% CI 1.836 - 2.818, P< 0.001). Publication bias and an influence of different cut-off values were not observed (all P> 0.05). In the pooled analyses of multivariate-adjusted HRs, the results suggested that pretreatment serum CEA may be an independent prognostic factor in gastric cancer (OS: HR 1.681, 95% CI 1.425 - 1.982; DSS: HR 1.900, 95% CI 1.441 - 2.505; DFS: HR 2.579, 95% CI 1.935 - 3.436).Conclusion/SignificanceThe meta-analysis based on the available literature supported the association of elevated pretreatment serum CEA levels with a poor prognosis for gastric cancer and a nearly doubled risk of mortality in gastric cancer patients. CEA may be an independent prognostic factor for gastric cancer patients and may aid in determining appropriate treatment which may preferentially benefit the CEA+ patients.

Diagnostic performance of CD66c in lung adenocarcinoma-associated malignant pleural effusion: comparison with CEA, CA 19-9, and CYFRA 21-1

Various tumour markers have been evaluated in malignant pleural effusions, but not CD66c. This study evaluated the diagnostic ability of CD66c in lung adenocarcinoma-associated malignant pleural effusions (LA-MPEs) and compared it with other known tumour markers. Forty-seven cases of LA-MPE and 52 cases of benign pleural effusions were collected. The levels of CD66c, CEA, CA 19-9, and CYFRA 21-1 were measured by enzyme immunoassay. The expression of CD66c, CEA, and CA 19-9 in cell blocks was measured by immunocytochemistry. CEA had the best diagnostic values, with a sensitivity of 87.2% and specificity of 92.3%. Both CD66c and CA 19-9 showed the highest specificity of 98.1%, with sensitivities of 63.8% and 55.3%, respectively. CYFRA 21-1 had a sensitivity of 83.0% and specificity of 76.9%. CEA combined with CA 19-9 reached a sensitivity of 91.5% and a specificity of 98.1%. The sensitivities of immunocytochemical staining for CD66c, CEA, and CA 19-9 were 72.5%, 75%, and 40%, respectively. CD66c showed a diagnostic performance comparable to CYFRA 21-1 and CA 19-9 by enzyme immunoassay. Immunocytochemical study showed that CD66c and CEA were more sensitive than CA19-9. Both studies support CD66c as a potential tumour marker to differentiate LA-MPE from benign effusions.

Expression of Molecular Markers and Risk Categorization in Pre-neoplastic and Non-neoplastic Cervical Lesions

Worldwide, cervical cancer is the fourth most common cancer in women [1]. It is preceded by many stages of pre-neoplastic lesions classified as Atypical squamous cells of undetermined significance (ASC-US) / Atypical squamous cells cannot exclude HSIL (ASC-H), Low-grade squamous intraepithelial lesion (LSIL), and High-grade squamous intraepithelial lesion (HSIL) (Bethesda, 2001). However, a large proportion of cases of epithelial malignancy arise from apparently normal cell and also from the LSIL where as many HSILs remain static for many years [2]. In this study, many important markers and epidemiological parameters Pap smears were examined to look for high risk cases of cervical non-malignant lesions. Methods: A total of 60 cases and controls were selected by colposcopic examination and smears were collected. Slides were used for immunohistochemical staining for four markers: p53, Ki-67, PCNA and CEA and PAP stain. All experimentally generated data were analyzed by Epi Info (TM) 3.5.3. Results: Expression of PCNA, p53, Ki-67 and CEA were significantly associated with both in Epithelial Cell Abnormalities (ECA) and Negative for Intraepithelial Lesion or Malignancy (NILM). Among HSIL, LSIL and ASCUS, HSIL was more common in Muslim women in comparison to Hindus. Whereas inflammatory lesions like RCC and RCCI were more common in Hindu population. Muslim women have a more normal Cytopathology smear than Hindus. Conclusions: Both the NILM and ECA cases carry risk for cancer conversion. Women with High grade Squamous Intraepithelial lesion or HSIL with PCNA, p53, Ki-67 and CEA strong positivity may have a higher risk of developing cervical cancer.

Inhibition of CEA release from epithelial cells by lipid A of Gram-negative bacteria.

A number of bacterial species, both pathogenic and non-pathogenic, use the human CEACAM family members as receptors for internalization into epithelial cells. The GPI-linked CEA and CEACAM6 might play a role in the innate immune defense, protecting the colon from microbial invasion. Previous studies showed that CEA is released from epithelial cells by an endogenous GPI-PLD enzyme. GPI-PLD activity was reported to be inhibited by several synthetic and natural forms of lipid A. We hypothesized that CEA engagement by Gram-negative bacteria might attenuate CEA release from epithelial cells and that this might facilitate bacterial colonization. We tested the hypothesis by examining the effect of Escherichia coli on CEA release from colorectal cancer cells in a co-culture experiment. A subconfluent monolayer culture of colorectal cancer cells (LS-180, Caco-2 and HT29/219) was incubated with E. coli. While there was a significant reduction in CEA secretion from LS-180 and HT29/219 cells, we found only a small reduction of CEA shedding from Caco-2 cells compared to the level from the untreated control cells. Furthermore, lipid A treatment of LS-180 cells inhibited CEA release from the cells in a dosedependent manner. Western blot analysis of total lysates showed that CEA expression levels in cells co-cultured with bacteria did not differ from those in untreated control cells. These results suggest that lipid A of Gram-negative bacteria might play a role in preventing the release of CEA from mucosal surfaces and promote mucosal colonization by bacteria.

Genomic profiling of patient-derived colon cancer xenograft models.

Recent evidence suggests that patient derived xenograft (PDX) models can maintain certain pathological and molecular features of the original disease. However, these characterizations are limited to immunohistochemistry or by tissue microarray analysis. We conducted a high-throughput sequencing of primary colon tumor and PDX has not been reported yet.Fresh primary colon cancer tissues that originate from surgery were implanted into the subcutaneous space of 6- to 8-week-old female BALB/c nu/nu or NOD/SCID mice and serially passaged in vivo. Ion AmpliSeq Cancer Hotspot Panel v2 (Ion Torrent) was used to detect frequent somatic mutations and similarity of molecular characteristics between the 10 patient tumors and matched PDX.Histologic and immunohistochemical analyses revealed a high degree of pathologic similarity including histologic architecture and expression of CEA, CK7, and CD20 between the patient and xenograft tumors. In 80% cases, all of the somatic mutations detected in primary tumor were concordantly detected in PDX models. However, 2 PDX models showed gained mutations such as PIK3CA or FBWX7 mutation.Ten patient-derived advanced colon cancer xenograft models were established. These models maintained the key characteristic features of the original tumors, suggesting useful tool for preclinical personalized medicine platform.

Abstract 4992: Zkscan3 facilitates invasion of colorectal cancer associated with ceacam5

Abstract Purpose: ZKSCAN3 is over-expressed in invasive colonic tumor cells and regulate the expression of several genes favoring tumor progression including integrin β4. We evaluated the role of ZKSCAN3 in invasive signaling pathway with stage 4 colorectal cancer (CRC). Materials and methods: 119 metastatic CRC (mCRC) patients who palliatively or curatively resected and 331 sporadic CRC patients who curatively resected from January 2004 to December 2009 were used to identify correlations between single-nucleotide polymorphism (SNP) rs733743 and clinically prognostic parameters. To validation of prior correlation, the genotypes of SNP identified by pyrosequencing using 81 patients (33 with mCRC and 48 CRC patients) and CRC cell lines. We then performed immunohistochemistry (IHC) and Western blot on 36 mCRC patients and 49 CRC patients. Up-regulated putative down-stream targets included genes contributing tumor aggressiveness and invasiveness; VEGF, cyclin D2, Akt, phospho-Akt, integrin β4, KRAS, CEACAM5, and integrin α5β1. To test biological utility assay, ZKSCAN3 mRNA was knocked down by ZKSCAN3 specific siRNA in metastatic CRC cells (LOVO) and ZKSCAN3 cDNA was overexpressed in the low-ZKSCAN3 expressed CRC cells (HCT116). Results: Wild-type alleles (GG) of ZKSCAN3 rs733743 was related with male dominant, family history of malignancy, high CEA concentration, and mCRC. The wild-type alleles of ZKSCAN3 rs733743 using tissue sample was related with lymphvascular invasion. ZKSCAN3 IHC positive tissue was related with lymphovascular invasion. ZKSCAN expression was higher in LOVO and SW620 cell lines which were derived from metastatic site. ZKSCAN3 high groups were related with CEA, integrin β4, VEGF, and AKT and ZKSCAN3 low groups were related with RAS, VEGF, AKT, and PAKT in metastatic tumor tissue. Therefore, ZKSCAN3 seemed to be related with CEA and integrin β4 to metastasis. Additional CEA expression was observed a significant decrease in ZKSCAN3 knockdowned LOVO cell (1.0 vs 0.89, P= 0.046) and invasiveness of the ZKSCAN3 overexpressed HCT116 cells was higher in CEA coated filters (75.6±6.0 vs 91.3±4.5, P=0.023). Conclusion: ZKSCAN3 is related with colorectal tumor progression and invasion. ZKSCAN3 overexpression tumor may facilitate metastasis of colorectal cancer associated with CEACAM5. Citation Format: Seon Ae Roh, Chan Wook Kim, Ka Hee Tak, Jin Cheon Kim. Zkscan3 facilitates invasion of colorectal cancer associated with ceacam5. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4992. doi:10.1158/1538-7445.AM2014-4992

189P - Diagnostic Performance of Cd66C in Lung Adenocarcinoma-Associated Malignant Pleural Effusion: Comparison with Cea, Ca 19-9, and Cyfra 21-1

ABSTRACT Aim: Various tumor markers have been evaluated to diagnose malignant pleural effusions, but not CD66c, a member of the CEACAM family. This study evaluated the diagnostic ability of CD66c in lung adenocarcinoma-associated malignant pleural effusions (LA-MPEs) and compared with other known tumor markers (CEA, CA 19-9, and CYFRA 21-1). Methods: Pleural effusions were collected from 47 patients with LA-MPE, and from 52 patients with benign conditions. The levels of CD66c, CEA, CA 19-9, and CYFRA 21-1 were measured by enzyme immunoassay. The expression of CD66c, CEA, and CA 19-9 in effusion cell blocks was measured by immunocytochemical staining. Results: The levels of all tumor markers were significantly higher in LA-MPE than in BPE (p Conclusions: CD66c showed a diagnostic performance comparable to CYFRA 21-1 and CA 19-9 by enzyme immunoassay. Immunocytochemical study showed that CD66c and CEA were more sensitive than CA19-9. Both studies support CD66c as a potential tumor marker to differentiate LA-MPE from benign effusions. Disclosure: All authors have declared no conflicts of interest.