A breast cancer cell microarray (CMA) as a rapid method to characterize candidate biomarkers

Abstract

Tissue microarrays (TMAs) have become an invaluable tool in cancer research to evaluate expression and subcellular localization of proteins in cells and tissues. As the catalogs of candidate biomarkers and therapeutic targets become more extensive, there is a need to characterize and validate these targets and biomarkers in cell lines as a primary biological system in research laboratories. Thus, cell microarrays (CMAs) are useful as a high-throughput screening tool. Here, we constructed a CMA containing 32 publicly available immortalized breast cell lines with the goal of creating a method to rapidly screen for antigens of interest in breast cancer research in a relatively easy, rapid and cost-effective manner. As proof of concept, we performed immunocytochemical staining of the HER2 receptor, as the status of this protein is relevant to breast cancer and has previously been reported for these cell lines. We observed a complete concordance of our staining with the published status of HER2 in these cell lines. In addition, we examined the expression of CD44, epithelial markers EpCAM and E-cadherin and tyrosine phosphoproteins. The labeling of these proteins correlates with the known biology of the cell lines. Our results demonstrate the utility of our method to screen for potential biomarkers and therapeutic targets in breast cancer and we suggest that CMAs be used as a general approach in breast cancer research.

Redox-Sensitive and Hyaluronic Acid-Functionalized Nanoparticles for Improving Breast Cancer Treatment by Cytoplasmic 17α-Methyltestosterone Delivery

“…Immunohistochemistry can be used to evaluate the expression and subcellular localization of proteins and other molecules in tissues. It can be a perfect tool in cancer therapy, where it is used as a method to verify the identity of tissue types, to categorize tumors, and to estimate the presence of specific molecules for different applications such as therapeutic or prognostic goals [21]. Herein, We used Immunohistochemistry to characterize CD44 expression levels on the surface of MCF-7 and BT-20 breast cancer cells, as shown in (Figure 11b) proved that MCF-7 was negative CD44, while BT-20 showed positive CD44 [22].…”

“…First, drug-loaded NPs were evaluated in terms of any alteration in morphology and MT state in different reducing environments. In addition, BT-20 cancer cell line with overexpression of CD44 and MCF-7 cancer cell line with low CD44 expression were used to confirm the selective binding affinity of HACSLA NPs to the HA receptors [21,22]. In vitro MTT, lactate dehydrogenase (LDH), Caspase-3 activity, Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and Rhodamine 123 (RH-123) assays were also accomplished to evaluate tumor-specific cytotoxicity and apoptosis induction due to MT-loaded HACSLA-NPs.…”

Progress in the clinical detection of heterogeneity in breast cancer

“…The concept was first introduced by Schena in 1995 , and is based on the ability of DNA to find and spontaneously bind its complementary sequence in a reversible way with high specificity . The technique can simultaneously measure the expression levels of thousands of genes and identify differentially expressed genes among different patients with cancer . Gene microarrays have been used to identify a number of differentially expressed genes .…”

“…The technique can simultaneously measure the expression levels of thousands of genes and identify differentially expressed genes among different patients with cancer . Gene microarrays have been used to identify a number of differentially expressed genes . As early as 2000, Perou and colleagues performed pattern analysis for gene expression in breast cancer using complementary DNA microarrays, initially discovering five major intrinsic gene signatures: luminal A, luminal B, Her‐2‐enriched, claudin‐low, and basal‐like .…”

Living Cell Microarrays: An Overview of Concepts

“…In the first reported study, a modified computer printer was used for cell patterning [ 67 ]. A variety of cell types, as well as isolated single-cells in a droplet of few picoliters can be printed at defined locations of a single microarray via inkjet technology [ 69 , 70 ]. The application of recently established piezoelectric non-contact nanoprinters provided us with the possibility to transfer very small amounts of living cells (1200 cells and fewer) to microscope slides and to subsequently cultivate and monitor the applied cells online [ 60 ].…”