Tumor Markers For Early Detection Research Articles

Vanadium-doped metal-organic framework@Znln2S4 core-shell heterojunction-attenuated photoelectrochemical immunoassay

Considering that cancer has become the second leading cause of death in humans, it is essential to develop an analytical approach that can sensitively detect tumor markers for early detection. We report an attenuated photoelectrochemical (PEC) immunoassay based on the organic-inorganic heterojunction 10MIL-88B(FeV)/ZnIn2S4 (10M88B(FeV)/ZIS) as a photoactive material for monitoring carcinoembryonic antigen (CEA). The 10M88B(FeV)/ZIS heterojunctions have excellent light-harvesting properties and high electrical conductivity, which are attributed to the advantages of both organic and inorganic semiconductors, namely, remarkable photogenerated carrier separation efficiency and long photogenerated carrier lifetime. Horseradish peroxidase (HRP) in the presence of H2O2 can catalyze 3,3′-diaminofenamide (DAB) producing brown precipitates (oxDAB), which is then loaded onto the 10M88B(FeV)/ZIS heterojunction to reduce the photocurrent and enable the quantitative detection of CEA. Under optimal conditions, the photocurrent values of the PEC biosensor are linearly related to the logarithm of the CEA concentrations, ranging from 0.01 ng mL−1 to 100 ng mL−1 with a detection limit (LOD) of 4.0 pg mL−1. Notably, the accuracy of the PEC biosensor is in agreement with that of the human CEA enzyme-linked immunosorbent assay (ELISA) kit.

Combinations of plasma cfDNA concentration, integrity and tumor markers are promising biomarkers for early diagnosis of non-small cell lung cancer

BackgroundCirculating cell-free DNA (cfDNA) concentration and integrity as noninvasive biomarkers play an important role in cancer diagnosis, prognosis and therapy monitoring. However, few studies have been conducted on the combination of plasma cfDNA concentration, integrity and tumor markers (CEA, CA125, NSE and CYFRA21-1) for cancer detection. Thus, the purpose of this study was to investigate the diagnostic value of combining plasma cfDNA concentration, integrity and tumor markers in early detection of non-small cell lung cancer (NSCLC). MethodsPlasma cfDNA concentration from 50 healthy controls and 84 NSCLC patients were assessed by quantitative real-time PCR of ALU repeated sequence. Plasma cfDNA integrity was calculated as the ratio of long to short fragments (ALU115/60). ResultsPlasma cfDNA concentration (ALU60 and ALU115) and integrity ALU115/60 were significantly higher in NSCLC patients with stage III/IV than in healthy controls (p = 0.0002, p < 0.0001, and p = 0.0093, respectively). The receiver operating characteristic (ROC) curve for discriminating NSCLC patients from healthy controls had an area under the curve (AUC) of 0.936 (95 % CI, 0.939–0.996). Moreover, the combination of plasma cfDNA concentration, integrity and tumor markers (CEA, CA125, NSE and CYFRA21-1) had higher diagnostic performance than either plasma cfDNA concentration alone, integrity alone or tumor markers alone, with sensitivity, specificity and AUC value of 94.05%, 90.00% and 0.968, respectively. These results demonstrated that the combination of plasma cfDNA concentration, integrity and tumor markers could significantly improve the diagnostic accuracy of NSCLC. ConclusionCombination of plasma cfDNA concentration, integrity and tumor markers is a promising biomarker for early diagnosis of NSCLC.

Tumor Marker in Early Detection of Malignancies in Type 2 Diabetes: A Case Series Analysis

Abstract The association between type 2 diabetes mellitus (T2DM) and malignancy has gained significant attention, with a heightened susceptibility to specific cancers. We present a comprehensive case series, documenting four distinctive cases illustrating the critical role of tumor markers in early cancer detection among patients with T2DM. Our findings emphasize the significance of vigilant surveillance and comprehensive patient care in the timely diagnosis and management of malignancies. The cases highlight the utility of tumor markers, particularly carbohydrate antigen 19-9, in the identification of pancreatic adenocarcinoma, lung cancer, and prostate cancer. These markers facilitated early interventions and subsequent tailored treatments, resulting in improved patient outcomes and extended survival. In addition, we underscore the challenges encountered in clinical practice, emphasizing the need for effective patient education and engagement in decision-making processes regarding their health. Although not currently recommended as routine screening tools, the potential lifesaving impact of tumor marker screenings in asymptomatic high-risk patients with T2DM warrants further consideration and research. This case series serves to underscore the critical role of tumor markers in uncovering potential malignancies at early stages, ultimately improving patient outcomes and quality of life.

Antígeno prostático específico en adultos de las parroquias urbanas de Cuenca - Ecuador

The objective of this research was to determine the values of prostate-specific antigen (PSA) in the blood serum of 1100 adults aged 40 to 80 who reside in the urban parishes of the city of Cuenca during 2018. The micro ELISA technique was used to determine PSA values, a glycoprotein secreted by the prostatic epithelium that allows sperm liquefaction and spermatozoa mobility. The results revealed that 8% of patients had values higher than 4 ng/ml, considered as the maximum limit of normality and present in prostatitis, benign prostatic hyperplasia, and prostate cancer. 50% of positive cases belonged to the age group of 70 to 80 years, finding statistical significance (p&lt;0.05) when related to variables such as age, family history, and sexual activity, highlighting the importance of establishing periodic control of this tumor marker for early detection.

An Overview: Genetic Tumor Markers for Early Detection and Current Gene Therapy Strategies

Genomic instability is considered a fundamental factor involved in any neoplastic disease. Consequently, the genetically unstable cells contribute to intratumoral genetic heterogeneity and phenotypic diversity of cancer. These genetic alterations can be detected by several diagnostic techniques of molecular biology and the detection of alteration in genomic integrity may serve as reliable genetic molecular markers for the early detection of cancer or cancer-related abnormal changes in the body cells. These genetic molecular markers can detect cancer earlier than any other method of cancer diagnosis, once a tumor is diagnosed, then replacement or therapeutic manipulation of these cancer-related abnormal genetic changes can be possible, which leads toward effective and target-specific cancer treatment and in many cases, personalized treatment of cancer could be performed without the adverse effects of chemotherapy and radiotherapy. In this review, we describe how these genetic molecular markers can be detected and the possible ways for the application of this gene diagnosis for gene therapy that can attack cancerous cells, directly or indirectly, which lead to overall improved management and quality of life for a cancer patient.

Diagnostic Value of Imaging Combined With Tumor Markers in Early Detection of Lung Cancer.

Objective: The objective of this research is to explore the diagnostic value of imaging plus tumor markers in the early detection of lung cancer.Methods: Sixty patients with lung cancer treated in our hospital from January 2018 to January 2019 were selected as group A. They were matched with 60 patients with benign lung disease as group B and 60 healthy subjects examined in our hospital as group C. The carcino-embryonic antigen (CEA), CYFRA21-1, and neuron-specific enolase (NSE) were assessed, and the diagnostic value of tumor markers plus imaging in lung cancer diagnosis was explored.Results: The CEA, CYFRA21-1, and NSE in group A were evidently superior to those in groups B and C, and those in group B were superior to those in group C (all P < 0.001). CEA had the highest sensitivity (56.7%), and NSE had the highest specificity (93.3%). The tumor markers plus imaging had the highest sensitivity for different types of lung cancer, and the sensitivity to early lung cancer (90%) was superior to other diagnostic methods (P < 0.05).Conclusion: The tumor markers plus imaging is of great significance in early lung cancer diagnosis and provides a reference for judging the pathological classification.

The Combined Diagnostic Accuracy of Serum Alpha Fetoprotein and Des-Gamma Carboxyprothrombin in Hepatocellular Carcinoma among Chronic Liver Disease Patients in Ilorin

Introduction :Chronic liver disease (CLD) is a disease of public health importance. CLD is defined as a clinical syndrome of liver disease lasting for at least six months with histology showing varying degree of hepatocellular necro-inflammation and fibrosis with or without neoplastic transformation. The disease is a spectrum that manifests initially as chronic hepatitis which may progress to liver cirrhosis and ultimately hepatocellular carcinoma (HCC). The current practice in the field of Gastroenterology has shifted from invasive methods of diagnosing HCC to non-invasive methods using tumor biomarkers. Various biomarkers of HCC have been proposed, but the largest body of evidence exists with alpha-fetoprotein (AFP). Most of the studies on the combined diagnostic accuracy of AFP and des-gamma-carboxyprothrombin (DCP) were done in other populations outside Nigeria. It is necessary to determine the combined diagnostic accuracy of the two tumor markers for early detection of HCC in North-central Nigeria. Materials and Methods: This study was a cross-sectional study and ethical clearance was obtained from the ethical and research committee of UITH, Ilorin. A total of 190 participants consisting of 125 cases and 65 healthy controls that were age and sex-matched were studied. Patients with extra-hepatic malignancies were excluded. The serum levels of AFP and DCP were determined using the enzyme-linked immunoassay (ELISA) technique. A detailed questionnaire was used to document the socio-demographic characteristics, clinical features as well as results of laboratory/radiologic parameters. Percutaneous liver biopsy was carried out on patients that were fit. Test of association between categorical variables was carried out using the Chi-Square Test. The sensitivity, specificity, positive and negative predictive values of the two tumor markers were determined by the area under curve (AUC) at various cut-off levels using the receiver operating characteristic (ROC) curve analysis. Statistical significance was set at p value EiTest MONO P-II kit) were used to assay AFP and DCP respectively. Liver biopsy needle (Menghini needle) was used to carry out liver biopsy. Results: Using a cut-off of 400 ng/ml, the sensitivity of serum AFP for diagnosing HCC was 51.3%. The specificity of AFP at the same cut-off was 87.8%. The positive and negative predictive values were 92.8% and 49.3% respectively. Using a cut-off of 7.5 ng/ml, the sensitivity of serum DCP for diagnosing HCC was 57.1%. The specificity of DCP at the same cut-off was 63.4%. The positive and negative predictive values were 76.2% and 41.9% respectively while the accuracy was 59.2%. The diagnostic accuracy of combined serum AFP and DCP for diagnosis of HCC in University of Ilorin Teaching Hospital, Ilorin was 64.9%. The sensitivity of combined serum AFP and DCP for diagnosing HCC was 55.6%. The specificity of combined serum AFP and DCP was 95.6%. The positive and negative predictive values were 96.2% and 52.3% respectively. Conclusion: Combining these two tumour markers does not significantly improve the diagnostic accuracy of HCC and chronic HBV remains a strong aetiological agent of HCC in UITH, Ilorin.

The Combined Diagnostic Accuracy of Serum Alpha Fetoprotein and Des-Gamma Carboxyprothrombin in Hepatocellular Carcinoma among Chronic Liver Disease Patients in Ilorin

Introduction :Chronic liver disease (CLD) is a disease of public health importance. CLD is defined as a clinical syndrome of liver disease lasting for at least six months with histology showing varying degree of hepatocellular necro-inflammation and fibrosis with or without neoplastic transformation. The disease is a spectrum that manifests initially as chronic hepatitis which may progress to liver cirrhosis and ultimately hepatocellular carcinoma (HCC). The current practice in the field of Gastroenterology has shifted from invasive methods of diagnosing HCC to non-invasive methods using tumor biomarkers. Various biomarkers of HCC have been proposed, but the largest body of evidence exists with alpha-fetoprotein (AFP). Most of the studies on the combined diagnostic accuracy of AFP and des-gamma-carboxyprothrombin (DCP) were done in other populations outside Nigeria. It is necessary to determine the combined diagnostic accuracy of the two tumor markers for early detection of HCC in North-central Nigeria. Materials and Methods: This study was a cross-sectional study and ethical clearance was obtained from the ethical and research committee of UITH, Ilorin. A total of 190 participants consisting of 125 cases and 65 healthy controls that were age and sex-matched were studied. Patients with extra-hepatic malignancies were excluded. The serum levels of AFP and DCP were determined using the enzyme-linked immunoassay (ELISA) technique. A detailed questionnaire was used to document the socio-demographic characteristics, clinical features as well as results of laboratory/radiologic parameters. Percutaneous liver biopsy was carried out on patients that were fit. Test of association between categorical variables was carried out using the Chi-Square Test. The sensitivity, specificity, positive and negative predictive values of the two tumor markers were determined by the area under curve (AUC) at various cut-off levels using the receiver operating characteristic (ROC) curve analysis. Statistical significance was set at p value EiTest MONO P-II kit) were used to assay AFP and DCP respectively. Liver biopsy needle (Menghini needle) was used to carry out liver biopsy. Results: Using a cut-off of 400 ng/ml, the sensitivity of serum AFP for diagnosing HCC was 51.3%. The specificity of AFP at the same cut-off was 87.8%. The positive and negative predictive values were 92.8% and 49.3% respectively. Using a cut-off of 7.5 ng/ml, the sensitivity of serum DCP for diagnosing HCC was 57.1%. The specificity of DCP at the same cut-off was 63.4%. The positive and negative predictive values were 76.2% and 41.9% respectively while the accuracy was 59.2%. The diagnostic accuracy of combined serum AFP and DCP for diagnosis of HCC in University of Ilorin Teaching Hospital, Ilorin was 64.9%. The sensitivity of combined serum AFP and DCP for diagnosing HCC was 55.6%. The specificity of combined serum AFP and DCP was 95.6%. The positive and negative predictive values were 96.2% and 52.3% respectively. Conclusion: Combining these two tumour markers does not significantly improve the diagnostic accuracy of HCC and chronic HBV remains a strong aetiological agent of HCC in UITH, Ilorin.

S1554 Unusual Mutation in Squamous Cell Carcinoma of Pancreas

INTRODUCTION: Pancreatic neoplasm is an aggressive malignant tumor of the digestive tract and a leading cause of death related worldwide. Squamous cell carcinoma of pancreas (SCCP) currently has an annual incidence rate of 0.02 cases per 100,000. The diagnosis is made only after excluding other neoplastic lesions of the pancreas that contain squamous epithelial components. CASE DESCRIPTION/METHODS: A 72 years old male with a history of diabetes mellitus, hypertension and a resected colorectal cancer that three years after resection, developed fatigue, unintentional weight loss, asthenia and anorexia. A contrast Abdominal CT revealed hepatomegaly suggestive of metastatic disease, intrahepatic and extrahepatic biliary duct dilation, and a pancreatic tail mass. CT guided biopsy of liver lesion showed metastatic squamous cells. PET scan identified a hypermetabolic, 3cm soft tissue mass in the left upper quadrant of the abdomen inseparable from the tail of the pancreas. Endoscopic ultrasound fine needle aspiration revealed a moderately differentiated SCCP. Next-generation sequencing came out with six genomic mutations, KRAS, FGFR1, NKX2-1, PBRM1, SPEN, and TP53. The patient was admitted to an oncologic unit where Nanoparticle albumin-bound paclitaxel-gemcitabine regimen was administered. However, due to complications including sepsis, and multi-organ failure, therapy was discontinued. Unfortunately, the patient succumbed to the latter. DISCUSSION: SCCP is a rare entity as squamous cells are naturally absent from the pancreas. However, these can be seen in conditions such as chronic pancreatitis, primary or metastatic carcinoma, and squamous metaplasia related to pancreatic or biliary duct stent placement. Squamous tumours are enriched for TP53 and KDM6A mutations, upregulation of the TP63 transcriptional network, hypermethylation of pancreatic endodermal cell-fate determining genes and have a poor prognosis. Thus, identifying patients who are at high risk to develop pancreatic cancer and who should undergo surveillance has to be a priority. In addition, there is a need to develop tumor markers for early detection, monitoring and treatment purposes. Precursor lesions might represent the opportunity to cure pancreatic neoplasia. With the integration of high throughput sequencing to the clinical practice, valuable information will be obtained. Consequently, providing better understanding of its pathogenesis, improving therapies alternatives and outcomes of pancreatic carcinomas.Figure 1.: Pancreas moderately differentiated Squamous Cell Carcinoma.Figure 2.: Multiple, hypermetabolic, large hepatic lesions, and a 3 cm soft mass in the left upper quadrant, inferior to the stomach and anterior to the spleen inseparable from the tails of pancreas.

A study for evaluating clinical relevance of circulating cell-free DNA in cervical cancer.

Recent techniques available for the detection of cervical cancer (CC) are highly invasive and costly, which makes it a rate-limiting step toward early diagnosis of this fatal disease. Evaluation of circulating cell-free DNA (ccfDNA) through liquid biopsy is a minimally invasive and cost-effective method that may serve as a unique tumor marker for early detection, treatment monitoring, the status of residual disease, and distant tumor metastasis in CC patients. In this study, initially, ccfDNA was measured in serum samples from 11 histopathologically proven cervix carcinoma patients and 8 controls. On successful screening, it was further extended to 2 more patients with a series of serum samples extracted at 3 different phases of the concurrent chemoradiotherapy (i.e., before, during, and after 6 months of follow-up). Agarose gel electrophoresis profile for ccfDNA of CC patients showed that of 11 patients, 4 patients had a comparatively higher tumor burden (ccfDNA) than the other 7 patients. Notably, during concurrent chemoradiotherapy, ccfDNA load disappeared and, after 6 months of follow-up, appeared back due to distant metastasis. Hence, we propose that this method could be an affordable and reliable way to diagnose/screen CC.

Clinical Impact of Oncomirs 221 and 222 on Breast Cancer Diagnosis

Background Dysregulation of miRNAs, non-coding RNAs of 18-25 ( ̴ 22nt), is a hallmark of malignancies among them is breast cancer. The present study aimed to investigate the expression levels of circulating oncomiRNAs (miRNA-221and miRNA-222) as a minimally non-invasive method for early detection of breast cancer as compared to tumor markers (CEA, CA15.3). Materials and methods MiRNA-221 and miRNA-222 expression levels were determined using quantitative real-time polymerase chain reaction (qPCR) in serum samples from three groups: primary breast cancer patients (n = 44), benign breast lesion patients (n = 25), and healthy individuals as control group (n = 19). Their diagnostic efficacy and relation with clinicopathological data were analyzed. Results MiRNA-221 and miRNA-222 expression and tumor markers reported significant increase in their mean levels in breast cancer group as compared to the benign breast lesions or control individuals. Among clinicopathological factors, miRs reported significant relation with pathological types, clinical staging, histological grading and hormonal status, while CEA and CA15.3 did not revealed significance with these factors. The diagnostic efficacy for investigated miRNAs was superior to tumor markers especially for detection of early stages and low grade tumors. Conclusion MiRNA-221 and miRNA-222 were superior over tumor markers for early detection of breast cancer especially those at high risk as primarybreast cancer patients with early stage or low grade tumors.

How Useful Are Tumor Markers in Detecting Metastases with FDG-PET/CT during Breast Cancer Surveillance?

Background: To assess the clinical usefulness of serum tumor markers for early detection of distant breast cancer recurrence using FDG-PET/CT. Methods: We retrospectively analyzed 561 consecutive patients who underwent surgery for invasive primary breast cancer and had increased tumor markers (CA 15-3 and CEA) after completion of locoregional therapy. FDG-PET/CT data were reviewed for all cases. CA 15-3 and CEA were evaluated both in a continuous and in a quartile (Q) distribution. The Wilcoxon rank-sum test and logistic regression models were used to evaluate the association between increased tumor marker values and the presence (and type) of distant metastases. Results: The median value of CA 15-3 was 35.0 U/mL (IQR, 29.5–43.0) in cases where no distant metastases were detected, and it was 58.9 U/mL (IQR, 40.0–108.0) in cases where metastases were detected (p < 0.001). The median value of CEA was 6.6 U/mL (IQR, 4.4–10.0) in cases of no metastases and 12.4 U/mL (IQR, 6.9–30.0) in cases of metastases (p < 0.001). Increased levels of both tumor markers (Q3 and Q4) were strongly associated with the presence of distant metastases. The association between CA 15-3 and bone/liver metastases was stronger compared with other types of metastases (p heterogeneity between odds ratios [ORs] = 0.03 for Q3 and <0.001 for Q4), while no relevant heterogeneity between ORs emerged for CEA. Conclusion: Increased tumor marker levels detected in asymptomatic breast cancer patients during adjuvant therapies and follow-up are significantly predictive of distant metastases identified on FDG-PET/CT.

Blood-based biomarkers for early detection of esophageal squamous cell carcinoma.

Esophageal squamous cell carcinoma (ESCC) is a common malignant tumor of the digestive system worldwide, especially in China. Due to the lack of effective early detection methods, ESCC patients often present at an advanced stage at the time of diagnosis, which seriously affects the prognosis of patients. At present, early detection of ESCC mainly depends on invasive and expensive endoscopy and histopathological biopsy. Therefore, there is an unmet need for a non-invasive method to detect ESCC in the early stages. With the emergence of a large class of non-invasive diagnostic tools, serum tumor markers have attracted much attention because of their potential for detection of early tumors. Therefore, the identification of serum tumor markers for early detection of ESCC is undoubtedly one of the most effective ways to achieve early diagnosis and treatment of ESCC. This article reviews the recent advances in the discovery of blood-based ESCC biomarkers, and discusses the origins, clinical applications, and technical challenges of clinical validation of various types of biomarkers.

Serum MiRNA-27a as potential diagnostic nucleic marker for breast cancer

Background Accumulating evidence reveals that microRNA 27a (miR 27a) is implicated in the pathogenesis of cancer. However, its diagnostic role in breast cancer (BC) still needs investigation. Materials and methods MiR 27a expression was assessed in serum samples from patients with primary BC (n = 100), benign breast lesions (n = 30) and control group served as healthy volunteers (n = 20) using quantitative real-time PCR. Results Both expression and mean rank of miR 27a and tumor markers among BC patients as compared to the other two groups. Clinicopathological characteristics showed significant relation with miRN 27a expression for clinical stage, histological grading, ER receptor and HER-2/neu. The diagnostic efficacy for miR 27a was superior to both tumor markers for early detection of BC especially high-risk BC groups. Conclusion Detection of miR 27a expression may serve as a potential sensitive minimally invasive molecular marker for early detection of primary BC.

The Stature of Human Thyroid Cancer Related to Over Expressed mir-221 and mir-146b

Purpose: The ability of miRNAs for biomarker tools was further built by recent research showing that cancerspecific miRNAs are traceable in cell-free blood fluids such as plasma and serum. In this study, the levels of miR-221 and miR146b in human’s peripheral blood as tumor markers for early detection of Papillary Thyroid Carcinoma [PTC] were investigated. Methods: A stem-loop quantitative reverse transcription-polymerase chain reaction [qRT-PCR] method was performed to determine the serum levels of miR-221 and miR-146b from 25 healthy controls, 25 PTC patients the day before thyroidectomy, 25 patients with benign nodules and 25 patients after thyroidectomy during the scheduled medical examinations. Results: There was statistical difference in the serum levels for miR-221 and miR-146b between the controls and the PTC patients. It also showed no statistical difference for miR-221 and miR-146b between controls and thyroidectomy patients. Conclusion: The overexpression of miR-221 and miR-146b could be potentially served as serum indicator biomarkers in the approach for Thyroid Papillary Carcinoma detection from benign nodules.

Abstract B36: Developing circRNA signatures as a biomarker for the early diagnosis of pancreatic carcinoma

Abstract Background: At the time of diagnosis, pancreatic ductal adenocarcinoma (PDAC) is typically already advanced and incurable. Current research has concentrated on finding tumor markers for early detection while the cancer is still localized and amenable to therapy, however, these markers remain elusive. The studies described focus on developing exonic circular RNAs (circRNA) as a novel set of diagnostic/prognostic biomarkers for PDAC. CircRNAs found in mammalian cells, are backsplice variants of transcripts that are derived from approximately 15% of actively transcribed genes. The prevalence, stability and cell-specific expression patterns of circRNAs suggest that they could be exploited as an indirect or surrogate readout of transcriptional activity in normal and diseased states. Both coding and non-coding RNAs are encapsulated within cytoplasmic endosomes, which are subsequently released as extracellular or circulating microvesicles called exosomes. Exosomes have become a promising research focus as a source for biomarkers. Objective: We are interested in elucidating whether aberrantly expressed genes in PDAC produce different types of circRNAs that become enriched in tumor-secreted exosomes. Hypothesis: Exosomal circRNA (exo-circRNA) expression patterns are potentially specific to different stages/types of PDAC and therefore can be used in disease sub-typing and prognosis. Methods: Exosomes were isolated from a normal pancreatic exocrine cell line (htert-HPNE) as well as three PDAC cell lines ranging from well to poorly differentiated, including PANC-1, BxPC3and MIAPaCa-2. The size and relative abundance of exosomes was quantified by transmission electron microscopy (TEM). The expression of common exosomal markers (CD63, CD9, CD81, and HSP70) and the PDAC exosomal marker glypican-1 (GPC-1) was evaluated by flow cytometry. RNA was purified from exosomes (exo-RNA) and the rRNA depleted samples were subject to circular RNA isolation. Exo-circRNA was used to construct RNA-Seq libraries. Sequencing of the generated libraries was performed on the Illumina Nextseq platform using 2x100 reads V1 chemistry at a targeted depth of 25 million paired end reads per library. The four read libraries were mapped to the human reference genome GRCh38.p3 using BWA-MEM, and analyzed using two bioinformatics platforms, “CIRI” and “find_circ”. Comparison of membership and expression levels was made between a normal cell line and well-, moderately- and poorly differentiated, PDAC cell lines. Results: Exosome size ranged from 20nm to 80nm. These structures demonstrated some diversity in size and marker expression when comparing cell lines. The smallest structures were observed from BxPC3 cell. Here, we show for the first time the presence of circRNAs in exosomes collected from PDAC cell lines. RNA-seq analyses revealed a number of interesting circRNA species that show cell line specificity. Preliminary examination of PANC-1 RNA-seq libraries from the ENCODE database identified over 800 circRNA isoforms from total cellular transcriptome. The number of circRNA isoforms for PANC-1 cells decreased when using an enriched exo-circRNA library for alignment to approximately 19 putative circRNA markers. Furthermore, circRNA isoforms for each of the cell lines examined were distinct. Interestingly, no circRNAs of genes known to be overexpressed in PDAC (such as K-RAS) were found in the fraction of exo-cricRNA for any of the cell lines tested. Impact: The studies described demonstrate that specific circRNAs can be readily extracted from the exosomes of conditioned media. We hope that this novel tool can be further developed to help to diagnose pancreatic carcinoma when it is amenable to surgical resection and/or chemotherapy, thereby reducing the mortality associated with this disease. Citation Format: Jessica Kalra, Keith Laderoute, Daniel Renouf, David Shaeffer, Marcel Bally. Developing circRNA signatures as a biomarker for the early diagnosis of pancreatic carcinoma. [abstract]. In: Proceedings of the AACR Special Conference on Noncoding RNAs and Cancer: Mechanisms to Medicines ; 2015 Dec 4-7; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2016;76(6 Suppl):Abstract nr B36.

Evaluation of the Diagnostic Performance of Soluble Human Leukocyte Antigen- G versus α-Fetoprotein for Hepatitis C Virus-Induced Hepatocellular Carcinoma

Background: Hepatocellular carcinoma (HCC) is the leading cause of death in HCV-related liver diseases and the third most common cause of cancer-related mortality worldwide. A reliable serum marker for early diagnosis of HCC is currently lacking. Objectives: The aim of this study was to identify the serum levels of soluble Human leukocyte antigen-G (sHLAG) in HCC and liver cirrhosis (LC) patients. As well, we aimed to estimate the diagnostic performance of sHLA-G for HCC by comparing its levels with the levels of Alpha fetoprotein (AFP). Subjects and methods: The study included 100 subjects divided into: 25 apparently healthy volunteers who served as healthy control subjects (group I), and 50 patients with untreated HCC on top of liver cirrhosis (group II) in addition to 25 cirrhotic patients (group III). HCC group was divided into two subgroups, 25 patients with AFP levels ≤ 200 ng/ml and 25 patients with AFP levels >200 ng/ml. All subjects were subjected to routine laboratory investigations plus detection of serum levels of both AFP and sHLA-G by Enzyme Immune Assay (EIA). Results: Blind parallel detection was conducted for sHLA-G and AFP. AFP was highly significantly increased in HCC and LC patients when compared to healthy controls (p=0.001). Also, there was a highly significant increase in AFP level in HCC patients when compared to cirrhotic ones (p=0.001). sHLA-G was highly significantly increased in HCC patients when compared to both healthy controls and cirrhotic patients (p=0.001). sHLA-G was not significantly increased in cirrhotic patients when compared to healthy controls (p=0.001). There was a significant strong positive correlations between sHLA-G and AFP in HCC patients (p<0.001) while weak negative correlation between sHLA-G and AFP was detected in cirrhotic group. The area under the receiver operating characteristic curve (ROC) was used to evaluate the diagnostic efficacies of both markers. The superiority of sHLA-G over AFP was more profound in the diagnosis of HCC [AUC: 0.993], in discriminating HCC from LC patients [AUC: 0.992] as well as in the diagnosis of HCC patients with AFP levels ≤ 200 ng/ml or in discriminating this HCC subgroup from LC patients [AUC: 0.986 and 0.984 respectively]. Conclusion: sHLA-G was found to be superior over AFP because of its higher AUC than that of AFP. As well, sHLA-G has better diagnostic performance with higher sensitivity and specificity than AFP. According to our data, sHLA-G could serve as a new efficient marker for early diagnosis of HCV-related HCC patients and to discriminate HCC from LC patients. Thus, measuring sHLA-G levels can help to reduce both false negative and false positive rates of AFP. Moreover, sHLA-G could have predictive value for tumorogenesis in HCV-related LC patients and could be used along with other tumor markers for early detection of malignant transformation.

Application of tumor markers SCC-Ag, CEA, and TPA in patients with cervical precancerous lesions.

To determine the potential clinical utility of tumor markers CEA, TPA, and SCC-Ag for early detection of cervical precancerous lesions. A case-control study was carried out on 120 women (46 patients with histologically confirmed cervical precancerous lesions and 74 healthy controls). The significance of serum selected tumor markers in early detection of cervical intraepithelial neoplasia (CIN) were assessed. Of the case group, the rates of CIN I, II, III, was 69.6%, 23.9%, and 6.5%, respectively. According to the manufacturer's cut-off values of 2 ng/ml, 5 ng/ml, and 70 U/ml for SCC-Ag, CEA and TPA tests, in that order, SCC-Ag test had a sensitivity of 13%, but CEA and TPA tests could not distinguish between case and control groups. The diagnostic sensitivities were highest at cut-off values of 0.55 ng/ml for SCC-Ag, 2.6 ng/ ml for CEA, and 25.5 U/ml for TPA which were 93%, 61%, and 50%, respectively. However, the area under the receiver operating characteristic curve was the largest for SCC-Ag (0.95 vs. 0.61 and 0.60 for CEA and TPA, respectively). Moreover, there was a highly significant direct correlation between SCC-Ag concentration and the degree of cervical precancerous lesions (r=0.847, p<0.001). The new cutoff of 0.5 for SCC-Ag test might be useful as a tumor marker in Iranian patients with CIN and it needs to be more evaluated by studies with larger populationa.

Absolute quantification of free tumor cells in the peripheral blood of gastric cancer patients.

Gastric cancer remains the third most common cancer in the world. Metastatic disease is a major cause of death in about half of the patients; therefore, early diagnosis is crucial for successful outcome. This study applied a sensitive method for the detection of circulating tumor cells using specific tumor markers for early detection. A total of 80 blood samples from 40 patients and 40 age-matched healthy controls were collected for the study. Circulating mRNA levels of two tumor markers, tumor endothelial marker 8 (TEM-8) and carcinoembryogenic antigen (CEA) were evaluated using absolute quantitative real-time PCR assay in the Stratagene Mx-3000P real-time PCR system. GAPDH was used to normalize the data. TEM-8 and CEA were detected in patients' blood more than in controls, 22/40 vs 9/40, P=0.005, and 30/40 vs 11/40, P=0.008, respectively. The mRNA level of these markers in patients was significantly higher in comparison to normal controls (P=0.018, 0.01). This panel showed an overall sensitivity of 64% and specificity of 73%. Statistical analysis for demographic variants did not show any significant differences. Both markers were detected more frequently and in significantly higher levels in blood samples of patients compared to samples from normal individuals. Copy number of CEA and TEM-8 mRNA, as detected by real-time quantitative PCR, appears to be a promising marker to evaluate the risk of tumor spread.

Expression of Ku86 and Presence of Ku86 Antibody as Biomarkers of Hepatitis B Virus Related Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) is a common disease and the third leading cause of cancer-related deaths worldwide. Level of the 82-kDa ATP-dependent DNA helicase II (Ku86) increases in some tumors, but its clinical use as a marker for HCC is rare. To examine the relationship between increases in Ku86 and the development of hepatitis B virus (HBV)-related HCC to define the relationship between Ku86 and HCC. Expression of Ku86 in tumor tissue, para-tumor tissue, and normal tissue was examined by immunohistochemistry, and Ku86 antibody titers in patient serum collected pre- and post-operatively were measured by ELISA. Long-term survival of the patients was also monitored. Ku86 staining in tumors was much stronger than in para-tumor and normal tissues. The expression of Ku86 was related to the tumor size, TNM stage, and tumor differentiation but not to gender, age, Child-Pugh score, tumor number, or α-fetoprotein levels. The long-term survival of patients with low Ku86 expression was longer. Patients with HCC had higher pre-operative Ku86 antibody levels. After surgical intervention, Ku86 antibody levels in patients with HCC declined significantly. Survival analysis showed that double-positive patients had the lowest survival rate, double-negative patients had the highest. Receiver operating characteristic curve analysis showed no significant difference between the AFP and Ku86 antibody. Multivariate analysis showed that Ku86 protein and Ku86 antibodies were independent prognostic factors of overall survival. Ku86 and Ku86 antibodies are promising tumor markers for early detection and prognosis prediction of HBV-related HCC.