RESUMO
OBJECTIVE: The present work aimed to investigate the potential utility of Sam68 protein as a prognostic marker in lung cancer. Then an electrochemical immunosensor is fabricated that is sufficiently sensitive to detect Sam68. RESULTS: Analysis of stage-specific Lung cancer microarray data shows that differential expression of Sam68 is associated with cancer stage and monotonically increases from early tumor stage to advanced metastatic stage. Moreover, the higher expression of Sam68 results in reduced survival of lung cancer patients. Based on these observations, an electrochemical immunosensor was developed for the quantification of Sam68 protein. The target protein was captured by the Anti-Sam68 antibody that was immobilized on the modified Glassy carbon electrode. The stepwise assembly process was characterized by cyclic voltammetry and electrochemical impedance spectroscopy. This fabricated immunosensor displayed good analytical performance in comparison to commercial ELISA kit with good sensitivity, lower detection limit (LOD) of 10.5 pg mL-1, and wide linear detection range from 1 to 5 µg mL-1. This method was validated with satisfactory detection of Sam68 protein in lung adenocarcinoma cell line, NCI-H23. Besides, spike and recovery assay reconfirm that the sensor can precisely quantify Sam68 protein in a complex physiological sample. CONCLUSION: We conclude Sam68 as a valuable prognostic biomarker for early detection of lung cancer. Moreover, we report the first study on the development of an electrochemical immunosensor for the detection of Sam68. The fabricated immunosensor exhibit excellent analytical performance, which can accurately predict the lung cancer patient pathological state.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/isolamento & purificação , Anticorpos/química , Técnicas Biossensoriais , Proteínas de Ligação a DNA/isolamento & purificação , Neoplasias Pulmonares/diagnóstico , Proteínas de Ligação a RNA/isolamento & purificação , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Adaptadoras de Transdução de Sinal/imunologia , Anticorpos/imunologia , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Limite de Detecção , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Fatores de Processamento de RNA/genética , Fatores de Processamento de RNA/isolamento & purificação , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/imunologiaRESUMO
An integrative approach is presented to identify grade-specific biomarkers for breast cancer. Grade-specific molecular interaction networks were constructed with differentially expressed genes (DEGs) of cancer grade 1, 2, and 3. We observed that the molecular network of grade3 is predominantly associated with cancer-specific processes. Among the top ten connected DEGs in the grade3, the increase in the expression of UBE2C and CCNB2 genes was statistically significant across different grades. Along with UBE2C and CCNB2 genes, the CDK1, KIF2C, NDC80, and CCNB2 genes are also profoundly expressed in different grades and reduce the patient's survival. Gene set enrichment analysis of these six genes reconfirms their role in metastatic phenotype. Moreover, the coexpression network shows a strong association of these six genes promotes cancer specific biological processes and possibly drives cancer from lower to a higher grade. Collectively the identified genes can act as potential biomarkers for breast cancer diagnosis and prognosis.
Assuntos
Biomarcadores Tumorais/genética , Neoplasias da Mama/genética , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/metabolismo , Neoplasias da Mama/mortalidade , Feminino , Humanos , Gradação de Tumores , Prognóstico , Análise de Sobrevida , TranscriptomaRESUMO
This work reports the design of a new electrochemical impedimetric immunosensor for the direct determination of ubiquitin-conjugating enzymes 2C (UBE2C), a potential diagnostic biomarker for breast cancer. The immunosensor was fabricated by immobilizing the capture anti-UBE2C antibody onto a polyaniline (PANI) modified glassy carbon electrode (GCE) through glutaraldehyde crosslinking. The assembly process of the immunosensor was examined using scanning electron microscopy, cyclic voltammetry, and electrochemical impedance spectroscopy. The fabricated immunosensor enabled the detection of recombinant human UBE2C in the range of 500 pg mL-1 to 5 µg mL-1. The limit of detection and limit of quantification were found to be 7.907 pg mL-1 and 26.356 pg mL-1, respectively. The diagnostic application of the fabricated immunosensor was explored for the analysis of breast cancer cell line MCF-7 cell extract. The immunosensor demonstrated high selectivity for UBE2C. The fabricated immunosensor also exhibited good reproducibility and storage stability.
RESUMO
Cancer is the second largest disease throughout the world with an increasing mortality rate over the past few years. The patient's survival rate is uncertain due to the limitations of cancer diagnosis and therapy. Early diagnosis of cancer is decisive for its successful treatment. A biomarker-based cancer diagnosis may significantly improve the early diagnosis and subsequent treatment. Biosensors play a crucial role in the detection of biomarkers as they are easy to use, portable, and can do analysis in real time. This review describes various biosensors designed for detecting nucleic acid and protein-based cancer biomarkers for cancer diagnosis. It mainly lays emphasis on different approaches to use electrochemical, optical, and mass-based transduction systems in cancer biomarker detection. It also highlights the analytical performances of various biosensor designs concerning cancer biomarkers in detail.
