Serum Samples Of Cancer Patients Research Articles

Comparative study of oxidative stress in cancer patients occupationally exposed to the mixture of pesticides.

Cancer isa leading cause of death worldwide, accounting for nearly 10 million deaths in 2020.Reviews indicated a positive relationship between exposure to pesticides and the development of cancers. In the present study, we have estimated the level of oxidative stress markers in serum samples of pesticide exposure and unexposed cancer patients as compared to normal control. We have found a significant decrease in peroxygenase (PON) and arylesterase (ARE) activity and substantial increases in homocysteine levels in both cancer groups. The level of heme biosynthesis rate-limiting enzymesdelta-aminolevulinic acid dehydratase(δ-ALA-D)also significantly decreased compared to control. The statistical comparison between the cancer groups does not show significant changes. We concluded the involvement of oxidative stress in carcinogenesis in both cancer group patients. However, more study is needed to put homocysteineas a novel marker for a variety of diseases on a single platform.

Evaluation of the JF5-based Aspergillus galactomannoprotein lateral flow device for diagnosing invasive aspergillosis in cancer patients.

Cancer patients are at heightened risk for invasive aspergillosis (IA), a condition associated with elevated mortality risk. The JF5-based Aspergillus Galactomannoprotein Lateral Flow Device (AspLFD) offers rapid point-of-care testing (POCT) for IA. This study evaluated the diagnostic performance of AspLFD in cancer populations. This retrospective study examined cancer patient bronchoalveolar lavage fluid (BALF) and serum samples collected between September 2021 and January 2023. Both AspLFD and galactomannan (GM) assays were conducted, and the results were analysed by two independent researchers. This study included 242 samples from 218 cancer patients, with 58 BALF and 184 serum samples. The overall agreement between AspLFD and GM assay results was 92.1%, with a kappa value of 0.552. AspLFD diagnosed proven/probable IA with a sensitivity and specificity of 91.7% and 95.3%, respectively, whereas GM exhibited sensitivity and specificity values of 83.3% and 93.7%, respectively. There were no statistical differences in the sensitivity and specificity between the two methods (P > 0.05). For serum analyses, AspLFD and GM exhibited similar sensitivity (66.7% vs. 66.7%, P > 0.05) and specificity (98.6% vs. 96.6%, P > 0.05) values. However, the sensitivity of the AspLFD was superior to the GM assay (100% vs. 88.9%) in BALF analyses but the difference was not statistically significant (P > 0.05), with no difference in specificity (83.7% vs. 83.7%, P > 0.05). In the solid-tumour cohort, both the AspLFD and GM assay exhibited high sensitivity (100% for both) and specificity (94.2% vs. 92.8%, P > 0.05). The AspLFD demonstrated good performance in diagnosing IA in cancer patients, especially those with solid tumours. The AspLFD is thus an alternative POCT, particularly when GM evaluations are not readily available.

Protein glycosylation in lung cancer from a mass spectrometry perspective.

Lung cancer is a severe disease for which better diagnostic and therapeutic approaches are urgently needed. Increasing evidence implies that aberrant protein glycosylation plays a crucial role in the pathogenesis and progression of lung cancer. Differences in glycosylation patterns have been previously observed between healthy and cancerous samples as well as between different lung cancer subtypes, which suggests untapped diagnostic potential. In addition, understanding the changes mediated by glycosylation may shed light on possible novel therapeutic targets and personalized treatment strategies for lung cancer patients. Mass spectrometry based glycomics and glycoproteomics have emerged as powerful tools for in-depth characterization of changes in protein glycosylation, providing valuable insights into the molecular basis of lung cancer. This paper reviews the literature on the analysis of protein glycosylation in lung cancer using mass spectrometry, which is dominated by manuscripts published over the past 5 years. Studies analyzing N-glycosylation, O-glycosylation, and glycosaminoglycan patterns in tissue, serum, plasma, and rare biological samples of lung cancer patients are highlighted. The current knowledge on the potential utility of glycan and glycoprotein biomarkers is also discussed.

Frequency shift Raman-based sensing of serum MicroRNA for ultrasensitive cervical cancer diagnosis

Cervical cancer is the most common gynaecological tumor. The development of a sensor for the ultrasensitive detection of cervical cancer is significant in guaranteeing its prognosis. Herein, we proposed a novel surface-enhanced Raman scattering (SERS) analysis platform using a frequency shifts-based sensing model for rapid and ultrasensitive microRNA (miRNA) assay. During the analysis process, miR-21 can be captured by the single-stranded DNA (ssDNA) modified on the platform which is complementary pairing with miR-21. The connection of miR-21 can lead to the variation of the molecular weight and result in the deformation extent of the Raman report molecule 6Thioguanine (6TG); thus, the peak at 1301 cm−1 due to the ring C–N stretches of 6TG shifts to lower frequency. The detection limit (LOD) of the proposed SERS analysis platform is as low as 8.32 aM. Moreover, the platform also has excellent specificity and repeatability, with the relative standard deviation (RSD) value of 6.53 %. Serum samples of cervical cancer patients and healthy subjects were analyzed via the platform and the accuracy of the detection results was verified by qRT-PCR, revealing that SERS results and qRT-PCR results have high homogeneity. Thus, the platform can serve as a potential tool for clinical diagnosis of cervical cancer.

Surface-enhanced Raman spectroscopy of the filtrate portions of the blood serum samples of breast cancer patients obtained by using 30 kDa filtration device

Raman spectroscopy is reliable tool for analyzing and exploring early disease diagnosis related to body fluids, such as blood serum, which contain low molecular weight fraction (LMWF) and high molecular weight fraction (HMWF) proteins. The disease biomarkers consist of LMWF which are dominated by HMWF hence their analysis is difficult. In this study, in order to overcome this issue, centrifugal filter devices of 30 kDa were used to obtain filtrate and residue portions obtained from whole blood serum samples of control and breast cancer diagnosed patients. The filtrate portions obtained in this way are expected to contain the marker proteins of breast cancer of the size below this filter size. These may include prolactin, Microphage migration inhabitation factor (MIF), γ-Synuclein, BCSG1, Leptin, MUC1, RS/DJ-1 present in the centrifuged blood serum (filtrate portions) which are then analyzed by the SERS technique to recognize the SERS spectral characteristics associated with the progression of breast cancer in the samples of different stages as compared to the healthy ones. The key intention of this study is to achieve early-stage breast cancer diagnosis through the utilization of Surface Enhanced Raman Spectroscopy (SERS) after the centrifugation of healthy and breast cancer serum samples with Amicon ultra-filter devices of 30 kDa. The silver nanoparticles with high plasmon resonance are used as a substrate for SERS analysis. Principal Component Analysis (PCA) and Partial Least Discriminant Analysis (PLS-DA) models are utilized as spectral classification tools to assess and predict rapid, reliable, and non-destructive SERS-based analysis. Notably, they were particularly effective in distinguishing between different SERS spectral groups of the cancerous and non-cancerous samples. By comparing all these spectral data sets to each other PLSDA shows the 79 % accuracy, 76 % specificity, and 81 % sensitivity in samples with AUC value of AUC = 0.774 SERS has proven to be a valuable technique for the rapid identification of the SERS spectral features of blood serum and its filtrate fractions from both healthy individuals and those with breast cancer, aiding in disease diagnosis.

Fabrication of a Low Cost Superhydrophobic Substrate for Surface Enhanced Laser-Induced Breakdown Spectroscopy and Its Utility through Identification of Electrolyte Variation for Oral Cancer Detection.

Ultratrace elemental detections from a limited volume of samples can offer significant benefits in biomedical fields. However, it can be challenging to concentrate the particles being analyzed in a small area to improve the accuracy of detection. Ring-like deposits on the edges of colloidal droplets are a vexing problem in many applications. Herein, we report ultratrace elemental detection using a superhydrophobic surface-enhanced laser-induced breakdown spectroscopy (SELIBS) substrate fabricated by laser ablation followed by a soft lithography technique. In this work, the SELIBS spectra on a superhydrophobic polydimethylsiloxane (PDMS) substrate replicated from a laser-patterned master Teflon substrate are investigated. This work highlights the application of this newly created superhydrophobic substrate for detecting trace elements in body fluids using SELIBS. The developed PDMS substrate was successfully adopted to investigate the electrolyte variation in serum samples of oral cancer patients and normal volunteers. Principal component analysis (PCA) and match-no-match analysis were used to distinguish the elemental variation in cancer and control groups.

The expression level of fibroblast growth factor gene in serum samples of lung cancer patients.

Lung cancer, one of the most deadly and dangerous types of cancer in the world, kills many men and women every year. Activation of fibroblast growth factor signals plays a role in the pathogenesis of several cancers, including lung cancer. Also, this factor may indicate prognosis and is related to the survival rate in patients with NSCLC. Therefore, this research investigated the level of fibroblast growth factor gene expression in the serum of people with lung cancer. In this research, 60 serum samples of healthy people and 60 serum samples of people with NSCLC were prepared, and personal and clinicopathological information of the studied people were collected by questionnaires. Then, plasma isolation, RNA extraction, cDNA synthesis, primers design, implementation, and changes in fibroblast growth factor gene expression in the serum of healthy and lung cancer patients were evaluated by the Real-time PCR method. REST software was used to analyze the results. The findings showed no significant difference in the expression of the fibroblast growth factor gene in the serum of people with the first to third stages of metastasis. However, in the serum of patients with the fourth stage of metastasis, the expression level of this gene was significantly decreased by 3.92 times compared to normal samples (P<0.05). According to the results of this study, it is possible to use the expression level of the fibroblast growth factor gene in people's serum to predict the metastasis stage of lung cancer.

Tailoring molecular recognition in predesigned multifunctional enzyme mimicking porphyrin imprinted interface for high affinity and differential selectivity; sensing etoposide in lung cancer patients

Nanozymes are cost-effective and robust but they lack specificity and selectivity, limiting their potential practical applications. Herein, molecularly imprinted polymers (MIPs) were grown in combination with multifunctional 5,10,15,20-tetrakis(4-hydroxyphenyl)-21H,23H-porphyrin (THPP) oxidase-like nanozyme to engineer THPP@MIP interface with high affinities and differential selectivity for structurally related target analytes. THPP nanozyme displayed a high level of predefined binding affinity for etoposide (ETO), and served as a predesigned functional monomer to rationally tailor the selectivity of THPP@MIP surface in the presence of different guest molecules. THPP nanozyme in combination with conventional monomers was imprinted on a portable and disposable cellulose paper matrix under UV light to create a UV-cured imprinted interface for optical detection of ETO. The THPP@MIP enzyme mimicking interface, having ETO specific and selective target recognition pockets, catalyzed the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) to generate visible blue oxidized TMB (oxTMB) without exogenous hydrogen peroxide (H2O2). The ETO binding on the THPP@MIP surface blocked the channels for TMB access to THPP cavities. The THPP@MIP sensor permitted to detect ETO in the linear range of 0.005–10 μg mL−1, with a limit of detection (LoD) of 0.002 μg mL−1, and showed a remarkable specificity and selectivity against other drug molecules. Furthermore, the THPP@MIP sensor successfully differentiated the serum samples of lung cancer patients and healthy volunteers. The obtained results were validated with standard High performance liquid chromatography-mass spectrometry (HPLC/MS) analysis of the serum samples. Additionally, ETO injection/infusion solutions and ETO-free serum samples were used to perform the matrix effect and recovery studies. This work demonstrates that molecular imprinting with predesigned, enzyme mimicking, high-affinity functional monomer can serve as a highly selective and specific universal interface for broad spectrum sensing applications in various analytical domains.

Exosomal circRNA RHOT1 promotes breast cancer progression by targeting miR-204-5p/ PRMT5 axis

BackgroundCircular RNA RHOT1 (circRHOT1) plays crucial roles in tumorigenesis by competing with microRNAs. It is largely abundant in tumor cell-derived exosomes. Meanwhile, cancer-derived exosomes participate in diverse biological processes. However, the expression patterns and functions of exosomal circRHOT1 in breast cancer remain unknown. This study is aimed to investigate and elucidate the exosomal circRHOT1/miR-204-5p/PRMT5 axis in breast cancer.MethodsThe exosomes derived from serum samples of breast cancer patients and breast cancer cell lines were characterized using transmission electron microscopy and Western blot. MTT, colony formation, wound healing, and transwell assays were utilized to analyze cell proliferation, migration, and invasion of breast cancer cells. Flow cytometry was used for apoptosis analysis. The bioinformatics method was employed to screen differentially expressed novel circRNAs and predict the microRNA targets of circRHOT1. Dual-luciferase reporter gene assays were performed to verify their direct interaction. Finally, Xenograft experiments were used to investigate the effect of exosomal circRHOT1 on tumor growth in vivo.ResultsCircRHOT1 exhibited significantly high expression in exosomes derived from the serum of breast cancer patients and breast cancer cell lines, which suggested its potential diagnostic value. Breast cancer-derived exosomes promoted the cell proliferation, migration, invasion, and epithelial-mesenchymal transition of breast cancer cells while inhibiting apoptosis. However, exosomes with downregulated circRHOT1 inhibited the growth of co-cultured cells. Mechanistically, circRHOT1 acted as a sponge of miR-204-5p and promoted protein arginine methyltransferase 5 (PRMT5) expression. Moreover, miR-204-5p inhibitor and pcPRMT5 could reverse the tumor suppressive effects mediated by circRHOT1-knockdown. Furthermore, treatment with exosomes derived from breast cancer cells with circRHOT1 knockdown attenuated tumor growth in tumor-bearing nude mice, which was accompanied by a reduction in PRMT5 expression and an enhancement of miR-204-5p expression.ConclusionThe exosomal circRHOT1 may promote breast cancer progression by regulating the miR-204-5p/PRMT5 axis. The current study strengthens the role of circRHOT1, miR-204-5p, and PRMT5 in breast cancer development and provides a potential treatment strategy for breast cancer.

B-365 Serum MAGE-A3 and MAGE-A4 Protein Levels as Potential Prognostic Biomarkers for Lung Cancer

Abstract Background Melanoma-associated antigen 3 (MAGE-A3) and MAGE-A4 are members of Melanoma Antigen Gene (MAGE) family, which have restricted expression to the testis and are abnormally expressed in cancer cells. MAGE-A3 and MAGE-A4 are overexpressed in lung cancer and have been shown to be associated with cancer progression and poor prognosis. Elevated MAGE-A4 protein level in lung cancer serum samples has been reported in the literature, however, it is unclear whether MAGE-A3 protein can be detected in serum samples of lung cancer patients. Methods We developed human MAGE-A3 and MAGE-A4 sandwich ELISA kits and used them to measure serum MAGE-A3 and MAGE-A4 protein levels in 21 lung cancer patients and 30 non-cancer controls. Results Serum MAGE-A3 protein was detected in all 21 lung cancer patients (range: 34 to 78871 pg/mL, mean: 5600 pg/mL) and was significantly elevated compared to non-cancer controls (range: 23–308 pg/mL, mean: 140 pg/mL) (P &amp;lt; 0.05). Serum MAGE-A4 protein was detected in 5 of 21 (24%) lung cancer patients (range: 280 to 35096 pg/mL, mean: 1819 pg/mL), and the samples with high serum MAGE-A4 concentration matched those with high serum MAGE-A3 concentration. Serum MAGE-A4 protein was not detectable in non-cancer controls. Conclusion Our findings suggest that serum MAGE-A3 and MAGE-A4 may serve as potential prognostic biomarkers for lung cancer. The detection of elevated serum MAGE-A3 and MAGE-A4 in lung cancer patients indicates their specificity for the disease. Larger studies with more samples, different cancer stages, and longer follow-up are needed to validate these results and establish the clinical utility of serum MAGE-A3 and MAGE-A4 levels as prognostic markers for lung cancer. If further validated, these biomarkers could aid in early diagnosis and better management of lung cancer.

An innovative and universal dual-signal ratiometric spectro-electrochemical imprinted sensor design for sandwich type detection of anticancer-drug, gemcitabine, in serum samples; cross validation via computational modeling

An innovative and universal imprinted sensor design for sandwich type detection of gemcitabine (GMT) in human serum samples is described. GMT is widely used in the treatment of different tumors, such as lung, ovarian, pancreatic, and breast cancer. The serum albumin-drug interaction was translated to design a multifunctional, ratiometric and dual mode silver nanoparticle based probe (BSA-Ag nanoprobe), as a read out system. Subsequently, polypyrrol imprinted drug receptor sites was engineered to selectively capture the GMT on the transducer surface. The GMT was sandwiched between imprinted receptor surface and BSA-Ag nanoprobe to generate the spectro-electrochemical signals. The formation of nanoprobe was confirmed through various characterization techniques, including X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, micro-Raman spectroscopy, Dynamic light scattering (DLS), and UV–Visible (UV–Vis) analysis, while each step of sensor fabrication was characterized via field emission scanning electron microscope (FE-SEM), Static water Contact angle measurements, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). Different variable parameters were optimized to improve the analytical performance of the sensor design. Under optimal conditions, spectro-electrochemical sensor permitted linear ranges between 1 and 200 μmol L−1 and 0.5–200 μmol L−1, with limits of detection (LOD) of 0.4 μmol L−1 and 0.15 μmol L−1 respectively. Furthermore, the designed sensor successfully differentiated the serum samples of lung cancer patients and healthy volunteers. The obtained results were validated with standard Liquid chromatography-mass spectrometry (LC/MS) analysis of the patients and healthy volunteer's serum samples. Lastly, density functional theory (DFT) and molecular docking calculations revealed the enhanced GMT binding capability of molecularly imprinted polypyrrole and molecular level interaction between the GMT and BSA, to validate the sandwich sensor design.

Study of Metallothionein Levels in the Serum of Patients with Malignant Tumour

Metallothionein (MT) is a low molecular weight intracellular protein, the primary function of which is to maintain the homeostasis of heavy metals in living organisms. Only a few research has been carried on the topic molecular mechanism of MT. Recent research points to its significant relationship to carcinogenesis, spontaneous mutagenesis, and the effectiveness of antitumor drugs. Increased level of MT is studied as a potential indicator of increased metabolic activity in cancer. Electrochemical methods are a suitable analytical tool for MT detection. Temperature-denatured blood serum samples of patients diagnosed with cancer were studied using electrochemical analysis (Brdička's reaction). The obtained voltammograms were evaluated as the area under the curve (AUC response). To compare normal and abnormal MT levels, control serum of healthy probands (n = 38) with an average MT amount of 2.2 µg L–1, median 2.1 µg L–1 was used. In all evaluated blood serum samples of cancer patients (n = 228), the average amount of MT was 22.7 µg L–1, and the median was 18.1 µg L–1. The difference (10-fold) between groups was highly statistically significant. The data obtained from the pilot experiment suggest that the levels of thiol compounds such as MT may have clinical potential in cancer patients.

A cohort study investigating the role of Bisphenol A in the molecular pathogenesis of breast cancer.

Breast cancer is an abnormal division of breast cells. Bisphenol A (BPA), an environmental toxicant, is identified as an emerging risk factor for breast cancer development. However, to the best of our knowledge, no previous study has investigated the BPA levels in breast cancer patients in Pakistan. The present study sought to explore the role of BPA in tumor growth among the Pakistani population. The levels of BPA were analyzed in the serum samples of breast cancer patients and controls by using HPLC. To elucidate the role of BPA to initiate tumorigenic events in breast tissue different biochemical assays along with expression analysis of tumor markers were performed. The level of BPA in the serum samples of breast cancer patients was significantly higher than control. Histological analysis of breast cancer tissue samples revealed distinct subtypes of tumor, such as ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC). There was a significant increase in ROS level while a significant decrease in the levels of superoxide dismutase (SOD) and catalase (CAT) enzymes in malignant breast tissue samples as compared to control tissue samples. We found upregulated expression of p53, ZEB1 and WNT1 genes at mRNA level in malignant breast tissue samples by 17 folds, 328 folds and 35 folds, respectively. p53 protein expression in malignant breast tissue samples was also enhanced at the translational level. Current findings suggest a relationship between BPA and the progression of breast cancer among the Pakistani population.

Procalcitonin assessment by using optical sensor terbium hydrochlorothiazide complex for early diagnosis of lung cancer

A simple, selective and sensitive spectrofluorimetric method has been reported in this paper for the assessment of procalcitonin (PCT) as a biomarker in serum samples of cancer patients by using optical sensor Tb3+-hydrochlorothiazide (Tb3+-HCTZ) complex. The method based on the quenching of the luminescence intensity of Tb3+-HCTZ complex by addition of different concentrations of PCT. The optical sensor was synthesized and characterized by absorption and fluorescence measurements. The performance of the designed sensor is determined through monitoring the quenching of the fluorescence intensity at 545 nm by (PCT) after excitation at 320 nm and pH = 7 in DMSO. The calibration of this method was performed in the concentration range of (3.37–100.62 ng/mL) with a limit of detection (LOD) and quantitation limit of detection (LOQ) are (0.038 and 0.114) ng/mL, respectively and a correlation coefficient of (0.98). This method was practical, simple and relatively free from interference effect. It was successfully applied to measure PCT in samples of human serum and from this method we can assess some biomarkers of cancer related diseases in human body.

Abstract 2168: Identifying a surrogate biomarker for neuropilin-2 axis in serum of prostate cancer patients

Abstract The purpose of this study is to investigate the correlation of neuropilin-2 (NRP2) and its ligand, vascular endothelial growth factor-C (VEGF-C), with treatment outcomes and resistance in prostate cancer patients. 166 prostate cancer patient serum samples were obtained from the Integrated Cancer Repository for Cancer Research at the University of Nebraska Medical Center. 11 patients were excluded for having a non-prostate genitourinary cancer or more than one concurrent cancer. Patients were categorized as untreated, primary treatment, remission, and secondary treatment based on when their serum sample was collected. Primary treatment indicated current treatment with first-line medical therapy, remission indicated those without evidence of disease after prostatectomy or first-line medical therapy, and secondary therapy indicated second-line treatment currently being given due to treatment-resistant disease. Other variables collected include age, race, ethnicity, prostate-specific antigen (PSA) measured at the time of diagnosis, PSA measured at the time of serum sample collection, Gleason score, and clinical stage using the American Joint Committee on Cancer (AJCC) tumor, node, metastasis (TNM) system. Finally, VEGF-C levels in patient serum were measured using a commercially available ELISA assay (R&amp;D System, Minneapolis, MN). The final data was analyzed using a linear regression model to compare VEGF-C levels between treatment groups, as well as VEGF-C level correlation with each of the other variables collected. Results: All patients were male with an age range of 47 to 85 years, with a mean age of 65.3 years. Most were white (n=135) and non-Hispanic (n=148). VEGF-C levels ranged from 341 pg/ml to 15,983 pg/ml, with a mean of 3,787 pg/ml. Of the 155 patients, 8 were untreated, 26 were in primary therapy, 66 were in remission, and 52 were in secondary therapy. Results showed no statistically significant correlation between VEGF-C levels and PSA at diagnosis, PSA at serum collection, Gleason score, or clinical stage. The most interesting difference between treatment groups was a higher VEGF-C level in remission (mean 63.48 pg/ml) versus secondary therapy (mean 55.31 pg/ml), although this was not statistically significant (p=0.089). Conclusions: Our results indicated that plasma VEGF-C alone is not a prognostic indicator for prostate cancer. Since our studies using cell culture and animal models of prostate cancer have indicated the role of VEGF-C and the NRP2 axis in promoting metastasis and therapy resistance in prostate cancer, it would be useful to determine both the VEGF-C and NRP2 levels in the plasma of the same patients and study whether their level together would predict the outcome of medical therapy. Our ongoing experiments to determine plasma NRP2 levels in this patient cohort should be able to test this hypothesis. Citation Format: Sarah A. Mullen, Dipanwita Das, Samikshan Dutta, Robin R. High, Lynette M. Smith, Benjamin A. Teply, Kaustubh Datta. Identifying a surrogate biomarker for neuropilin-2 axis in serum of prostate cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2168.

Circulating miRNA as a Biomarker in Oral Cancer Liquid Biopsy.

Oral cancer is currently challenging the healthcare system, with a high incidence among the population and a poor survival rate. One of the main focuses related to this malignancy is the urge to implement a viable approach for improving its early diagnosis. By introducing the use of liquid biopsy and the identification of potential biomarkers, aiming for a noninvasive approach, new advancements offer promising perspectives in the diagnosis of oral cancer. The present review discusses the potential of circulating miRNAs as oral cancer biomarkers identified in body fluids such as serum, plasma, and saliva samples of oral cancer patients. Existing results reveal an important implication of different miRNA expressions involved in the initiation, development, progression, and metastasis rate of oral malignancy. Liquid biomarkers can play a crucial role in the development of the concept of personalized medicine, providing a wide range of clinical applications and future targeted therapies.

Abstract P2-26-22: Elevated Expression of High Mannose Glycans in Tissue and Serum Samples of Breast Cancer Patients

Abstract Glycobiology has proved to be a new frontier in oncology research in recent years. One sector of glycobiology involves examining glycan expression of cancerous cells. Glycans are complex carbohydrates found on cell surfaces, and patterns of aberrant glycan expression, such as increased sialylation and fucosylation, have been proposed as potential novel biomarkers of cancer tumorigenesis and metastasis. However, to our knowledge, there is little literature examining the differences in expression of high mannose glycans, a subtype of N-linked glycans, which is the focus of this study. Thirty-nine samples of malignant and surrounding benign tissue from breast cancer patients were collected from the Prisma Health Cancer Institute Biorepository. Additionally, forty malignant serum samples were collected—ten of these samples were from the same patients included in the tissue samples. Serum samples from ten healthy volunteers were also obtained as controls. The malignant samples included specimens from breast cancer staged I through IV at the time of the patients’ diagnosis. Samples were analyzed using MALDI-TOF mass spectroscopy for the presence of glycans by Emory Glycomics and Molecular Interactions Core. The known masses of sixty-one unique glycans were then compared to the spectra of the samples analyzed to determine the type of glycans present, and the relative amount of each glycan present was determined using the area under the curve in the spectral data. Data analysis revealed the means of the area under the curve for high mannose glycans in the malignant and normal tissue samples to be 247.18 and 163.6 respectively; the t-test established the difference in these means to be statistically significant (p-value, 0.0138). This finding was consistent in serum samples; the mean area under the curve for malignant and normal serum was 1650.34 and 376.26 respectively (p-value, &amp;gt;.0001). Furthermore, there were differences in expression of high mannose glycans between stages of breast cancer, with expression in stage III being significantly greater than expression in stage II in both serum (p-value, .0097) and tissue (p-value, .0003) samples. The results of this analysis suggest a higher expression of high mannose glycans to be a promising biomarker for breast cancer. This could lead to important clinical applications such as being a helpful distinction in obtaining negative margins during tumor resection, as well as providing a non-invasive diagnostic method. Future research could include expanding the sample size and comparing the glycan composition of the samples to a library specific for high-mannose type glycans, with the goal of establishing a signature high-mannose expression pattern in each stage of cancer. Citation Format: Alex Kesic, Avery Funkhouser, Jonah Shealy, Julie Martin, W. Larry Gluck, William J. Edenfield, Anna Blenda. Elevated Expression of High Mannose Glycans in Tissue and Serum Samples of Breast Cancer Patients [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-26-22.

Experimental study on the suppressive effect of B3GNT3 on the apoptosis of lung adenocarcinoma cells and its application in early screening for lung adenocarcinoma.

Lung adenocarcinoma, as a common histological type in lung cancer, the overall survival is very low, and the prognosis is poor because it is difficult to find and easily recurs. Therefore, this study aimed to explore the role of the secreted protein beta-1,3-N-acetylglucosaminyltransferase 3 (B3GNT3) in the development of lung adenocarcinoma and to evaluate its potential significance for early clinical biomarker screening. The mRNA expression profiles of patients with lung adenocarcinoma and normal controls were analyzed via The Cancer Genome Atlas (TCGA) database. Serum samples of clinical lung cancer patients and healthy people were obtained, and the differences in B3GNT3 expression in different stages of lung adenocarcinoma and in healthy tissues were compared. Kaplan-Meier (K-M) curves were drawn to clarify the influence of high and low expression of B3GNT3 on the prognosis of patients. Peripheral blood samples from patients with lung adenocarcinoma and healthy people were obtained clinically, and receiver operating characteristic (ROC) curves were drawn to clarify the sensitivity and specificity of B3GNT3 expression for the diagnosis of lung adenocarcinoma. Lung adenocarcinoma cells were cultured in vitro, the expression of B3GNT3 was knocked down by lentivirus infection. The expression of the apoptosis-associated genes was detected by reverse transcription-polymerase chain reaction (RT-PCR). The secreted protein B3GNT3 is significantly differentially expressed in the serum of patients with lung adenocarcinoma versus normal controls. Subgroup analysis according to lung adenocarcinoma clinical stage showed that the higher the clinical stage of lung adenocarcinoma was, the higher the B3GNT3 expression. Enzyme-linked immunosorbent assay (ELISA) revealed that B3GNT3 expression was significantly increased in the serum of patients with lung adenocarcinoma and significantly decreased after surgery. By inhibiting programmed cell death-ligand 1 (PD-L1), the level of apoptosis was significantly increased and the proliferative capacity was significantly inhibited. In contrast, the level of apoptosis was significantly increased and the proliferation ability was significantly inhibited after simultaneous overexpression of B3GNT3 and inhibition of PD-L1. High expression of the secreted protein B3GNT3 in lung adenocarcinoma is closely related to prognosis and can serve as a potential biological marker for the early screening of lung adenocarcinoma.

Abstract P2-23-18: Analysis of Serum Galectin Profiles by Breast Cancer Subtypes and Patient Characteristics

Abstract Background: Galectins are sugar binding proteins that play a role in adhesion, apoptosis, immune regulation, and many other cellular processes. There are three different classes of galectins, and they are all found intracellularly and secreted in serum. Galectins have an established relationship with breast cancer, and galectin-3 has been repeatedly associated with cancer cell survival and tumor progression. Most research regarding breast cancer and galectins has used immunohistochemistry (IHC) as a method for identifying galectin expression in breast cancer tissue samples. However, little research has identified a relationship between serum galectin concentrations and breast cancer subtypes. Breast cancer is characterized both by histological and molecular subtype, and this identification often directs treatment decisions. The current method for identifying breast cancer subtypes is through invasive biopsy and the subsequent use of IHC. This project characterized serum galectin concentrations in breast cancer patients to determine if they changed based on different histological and molecular subtypes, cancer stage, and other patient characteristics. Our goal was to evaluate a potential noninvasive means of breast cancer subtype identification and to identify potential therapeutic targets. Methods: One-hundred breast cancer patient serum samples were studied using Enzyme-linked Immunosorbent Assays (ELISAs) to determine concentrations of galectins -1, -3, -7, and -9. Results were compared using many patient characteristics, including breast cancer subtype, stage, and patient history (smoking status and treatment). Statistical Analysis was performed using ANOVA, Student’s t-test, and the Wilcoxon Method. Results: The concentrations of galectins -1, -3, and -9 in breast cancer patients were all significantly higher than those of healthy controls (Table 1), which is consistent with previous studies. The concentration of galectin-3 was significantly higher in invasive lobular carcinoma samples (N, 6; mean, 13.04 ng/mL) compared to invasive ductal carcinoma samples (N, 80; mean, 9.93 ng/mL; p-value, 0.0428). Galectin-3 concentrations were significantly lower in samples from breast cancer patients who received chemotherapy (N, 9; mean, 9.22 ng/mL) versus those who did not (N, 7; mean, 14.60 ng/mL; p-value, 0.0228). Concentrations of galectin-1 were found to increase by stage (p-value, 0.0031), with significant differences between samples from patients with stage I breast cancer (N, 45; mean, 19.02 ng/mL) and both stage II (N, 40; mean, 24.15 ng/mL; p-value, 0.0127) and stage III (N, 12; mean, 28.37 ng/mL; p-value, 0.0026) disease. Conclusions: Our findings suggest a potential use for serum galectin-3 concentrations as a non-invasive means for breast cancer histological subtype differentiation and more refined diagnosis. In addition, our results suggest a possible use of galectin-1 concentrations for more accurate cancer staging. Further research could explore the relationship of serum galectin concentrations and other breast cancer subtypes. Future studies could also analyze how galectin concentrations change with patient treatment status and determine the potential of galectins as specific targets for breast cancer treatment. Table 1: Galectin Concentrations in Breast Cancer vs Healthy Controls Citation Format: Jonah Shealy, Alex Kesic, Avery Funkhouser, Julie Martin, W. Larry Gluck, w. Jeffery Edenfield, Anna Blenda. Analysis of Serum Galectin Profiles by Breast Cancer Subtypes and Patient Characteristics [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-23-18.

Sensitive determination of prostate-specific antigen with graphene quantum dot-based fluorescence aptasensor using few-layer V2CTx MXene as quencher

A novel fluorescence aptasensor of prostate-specific antigen (PSA) was established using few-layer vanadium carbide (FL-V2CTx) nanosheet as a quencher. First, FL-V2CTx was prepared by the delamination of multi-layer V2CTx (ML-V2CTx) with tetramethylammonium hydroxide. The aptamer-carboxyl graphene quantum dots (CGQDs) probe was prepared by combining the aminated PSA aptamer and CGQDs. Then, the aptamer-CGQDs were absorbed onto the surface of FL-V2CTx by hydrogen bond interaction, which led to the decrease in fluorescence of aptamer-CGQDs due to photoinduced energy transfer. After addition of PSA, PSA-aptamer-CGQDs complex was released from FL-V2CTx. The fluorescence intensity of aptamer-CGQDs-FL-V2CTx with PSA was higher than that without PSA. The FL-V2CTx-based fluorescence aptasensor provided a PSA detection linear range from 0.1 to 20 ng mL−1 with detection limit of 0.03 ng mL−1. The ΔF value of fluorescence intensities for aptamer-CGQDs-FL-V2CTx with and without PSA was 5.6, 3.7, 7.7, and 5.4 times of ML-V2CTx, few-layer titanium carbide (FL-Ti3C2Tx), ML-Ti3C2Tx and graphene oxide aptasensors, respectively, indicating the advantage of FL-V2CTx. The aptasensor had high selectivity for PSA detection compared with some proteins and tumor markers. This proposed method had convenience and high sensitivity for PSA determination. The determination results of PSA in human serum samples using the aptasensor were consistent with those by chemiluminescent immunoanalysis. The fluorescence aptasensor can be successfully applied for PSA determination in serum samples of prostate cancer patients.