Multilectin Affinity Chromatography for Characterization of Multiple Glycoprotein Biomarker Candidates in Serum from Breast Cancer Patients

Abstract Background: Glycoproteins are often associated with cancer and are important in serum studies, for which glycosylation is a common posttranslational modification. Methods: We used multilectin affinity chromatography (M-LAC) to isolate glycoproteins from the sera of breast cancer patients and controls. The proteins were identified by HPLC–tandem mass spectrometry (MS/MS) analysis of the corresponding tryptic digests. We used the FuncAssociate Gene Ontology program for association analysis of the identified proteins. Biomarker candidates in these groups were comparatively quantitated by use of peak area measurements, with inclusion of an internal standard. We analyzed data for concordance within the ontology association groups for vector of change with the development of breast cancer. Results: Detection of the known low-concentration biomarker HER-2 (8–24 μg/L) enabled us to establish a dynamic range of 106, relative to the amount of albumin, for the depletion step. We then used ELISA to confirm this range. Proteins associated with lipid transport and metabolism, cell growth and maintenance, ion homeostasis, and protease inhibition were found to be differentially regulated in serum from women with breast cancer compared with serum from women without breast cancer. Conclusions: M-LAC for isolation of the serum glycoproteome, coupled with liquid chromatography–MS/MS and the use of gene ontology associations, can be used to characterize large panels of candidate markers, which can then be evaluated in a particular patient population.

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Next-Generation Sequencing of MicroRNAs for Breast Cancer Detection

Journal of Biomedicine and Biotechnology ◽

10.1155/2011/597145 ◽

2011 ◽

Vol 2011 ◽

pp. 1-7 ◽

Cited By ~ 49

Author(s):

Qian Wu ◽

Zuhong Lu ◽

Hailing Li ◽

Jiafeng Lu ◽

Li Guo ◽

...

Keyword(s):

Breast Cancer ◽

Gene Ontology ◽

Cancer Patients ◽

Cancer Detection ◽

Target Genes ◽

Breast Cancer Detection ◽

Gene Ontology Analysis ◽

Breast Cancer Patients ◽

Mirna Profiling ◽

Serum Mirnas

It is reported that different microRNA (miRNA) profiles can be detected in the blood of cancer patients. We investigated that whether the key serum miRNAs could discriminate patients with and without breast cancer. This study was divided into three parts: (1) miRNA marker discovery using SOLiD sequencing-based miRNA profiling on cancerous and adjacent noncancerous breast tissue of one breast cancer patient; (2) marker selection and validation by real-time PCR on a small set of serum; (3) gene ontology analysis of the key miRNA target genes. Of genome-wide tissue miRNA expression analysis, five miRNAs were found to be altered more than fivefold by SOLiD sequencing (i.e., miR-29a, miR-23a, miR-23b, miR-192, and miR-21). All the five miRNAs were validated on the 20 breast cancer patients and 20 controls. miR-29a and miR-21 were significantly increased in the serum of breast cancer patients (). Gene ontology analysis of the target genes revealed enrichment for special biological process categories, that is, signal transduction, development, apoptosis, cell proliferation, and cell adhesion. SOLiD sequencing provides a promising method for cancer-related miRNA profiling. Serum miRNAs may be useful biomarkers for breast cancer detection.

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INVESTIGATING THE RELATIONSHIPS BETWEEN EXPRESSED CANCER RELATED GENES AND SURVIVAL OF PATIENTS WITH BREAST CANCER

FUDMA Journal of Sciences ◽

10.33003/fjs-2021-0502-553 ◽

2021 ◽

Vol 5 (2) ◽

pp. 327-333

Author(s):

Chinenye Nworah ◽

Bashir Sule

Keyword(s):

Breast Cancer ◽

Cancer Patients ◽

Dynamic Range ◽

Pearson Correlation ◽

Expression Patterns ◽

Parametric Models ◽

Normal Model ◽

Breast Cancer Patients ◽

Expression Levels ◽

Log Normal

Cancer stem cells are regulated by complex interactions with the components of the tumor microenvironment through networks of Cytokins and growth factors. These interactions are mediated by group of proteins and microRNAs (miRs), which are expressed or repressed. These expression levels are critical for cancer stem cell formation and expansion, enabling the promotion of tumor cell proliferation and migration, as well as for the survival of cancer recurrence and patient survival. Micro array and RNA deep sequencing (RNA-seq) provide tools with ability to generate transcriptome information, deciphering global gene expression patterns and quantifying a large dynamic range of expression levels. In this study 94 breast cancer patients were investigated based on miR and mRNA expression levels in which WDR1, APC and AKAP13 genes were identiﬁed as genes that play important role in the survival of patients and these genes differed signiﬁcantly with respect to survival of patients. We used the Pearson correlation to identify the over-expressed and under-expressed genes. We demonstrated that parametric survival models can be used to model outcomes for breast cancer, and for our dataset the log-normal model demonstrated the best ﬁt compared to other parametric models. Through the use of log-normal model, we examined how each of the identiﬁed genes inﬂuence the survival of breast cancer patients.

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An Ultrasensitive Routine LC-MS/MS Method for Estradiol and Estrone in the Clinically Relevant Sub-Picomolar Range

Journal of the Endocrine Society ◽

10.1210/jendso/bvaa047 ◽

2020 ◽

Vol 4 (6) ◽

Cited By ~ 2

Author(s):

Bjørn-Erik Bertelsen ◽

Ralf Kellmann ◽

Kristin Viste ◽

Anne Turid Bjørnevik ◽

Hans Petter Eikesdal ◽

...

Keyword(s):

Breast Cancer ◽

Aromatase Inhibitor ◽

Cancer Patients ◽

Aromatase Inhibitors ◽

Internal Standard ◽

Postmenopausal Breast Cancer ◽

Reference Ranges ◽

Breast Cancer Patients ◽

Endocrine Society ◽

Lh Releasing Hormone

Abstract Background Current analytical routine methods lack the sensitivity to monitor plasma estrogen levels in breast cancer patients treated with aromatase inhibitors. Such monitoring is warranted for premenopausal patients treated with an aromatase inhibitor and an LH-releasing hormone analogue in particular. Therefore, we aimed to develop a routine tandem mass spectroscopy combined with liquid chromatography (LC-MS/MS) method for estradiol (E2) and estrone (E1) for use in the sub-picomolar range. Method Calibrators, quality controls (QC), or serum samples were spiked with isotope-labeled internal standard and purified by liquid-liquid extraction. The reconstituted extracts were analyzed by LC-MS/MS in negative electrospray ionization mode. QCs at 6 levels made from pooled patient sera were used to validate the accuracy, sensitivity, and precision of the method. Results We achieved limits of quantification of 0.6 pmol/L (0.16 pg/mL) for E2 and 0.3 pmol/L (0.07 pg/mL) for E1. The coefficient of variation was below 9.0% at all QC levels for E2 (range, 1.7-153 pmol/L), and below 7.8% for E1 (range, 1.7-143 pmol/L). The method is traceable to the E2 reference standard BCR576. Reference ranges for E2 and E1 in healthy, postmenopausal women were obtained, for E2: 3.8 to 36 pmol/L, for E1: 22 to 122 pmol/L. We measured and confirmed ultra-low E2 and E1 concentrations in sera from patients on the aromatase inhibitors letrozole or exemestane. Conclusion This ultrasensitive LC-MS/MS method is suitable for routine assessment of serum E1 and E2 levels in breast cancer patients during estrogen suppression therapy. The method satisfies all requirements for measurement of E2 in the clinical setting as stated by the Endocrine Society in 2013. Precis We report an ultrasensitive LCMS/MS routine assay that measures pretreatment and suppressed levels of estradiol/estrone during aromatase inhibitor treatment of postmenopausal breast cancer patients.

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