High‐content cell death imaging using quantum dot‐based TIRF microscopy for the determination of anticancer activity against breast cancer stem cell

Abstract

We report a two color monitoring of drug-induced cell deaths using total internal reflection fluorescence (TIRF) as a novel method to determine anticancer activity. Instead of cancer cells, breast cancer stem cells (CSCs) were directly tested in the present assay to determine the effective concentration (EC ) values of camptothecin and cisplatin. Phosphatidylserine and HMGB1 protein were concurrently detected to observe apoptotic and necrotic cell death induced by anticancer drugs using quantum dot (Qdot)-antibody conjugates. Only 50-to-100 breast CSCs were consumed at each cell chamber due to the high sensitivity of Qdot-based TIRF. The high sensitivity of Qdot-based TIRF, that enables the consumption of a small number of cells, is advantageous for cost-effective large-scale drug screening. In addition, unlike MTT assay, this approach can provide a more uniform range of EC values because the average values of single breast CSCs fluorescence intensities are observed to acquire EC values as a function of dose. This research successfully demonstrated the possibility that Qdot-based TIRF can be widely used as an improved alternative to MTT assay for the determination of anticancer drug efficacies.

The Inhibitory Effect of Curcumin on Hypoxia Inducer Factors (Hifs) as a Regulatory Factor in the Growth of Tumor Cells in Breast Cancer Stem-Like Cells

“…Recent studies have indicated that existing CSCs in solid tumors lead to chemotherapy resistance; therefore, CSCs should be eliminated to overcome drug resistance [15]. CSCs are a limited subpopulation with specific properties like self-renewal that are detected in several malignancies such as melanoma, breast, prostate and colon [16,17]. Several studies have focused on isolating CSCs according to surface markers in order to develop new anticancer agents for targeted chemotherapy [17,18].…”

Quantum Dot Research in Breast Cancer: Challenges and Prospects

“…Advances in delivery systems using quantum dots highlight their potential in nanomedicine, with modifications that enhance biocompatibility and efficacy [44,45,[52][53][54][55]. In targeted and co-delivery systems, quantum dots play a crucial role in advanced therapy, merging with biomolecules for precise therapeutic delivery, marking significant progress in medical nanotechnology [2,3,12,18,19,[21][22][23]56,57].…”

“…Quantum dot-based total internal reflection fluorescence (TIRF) microscopy was utilized to monitor drug-induced cell death in breast cancer stem cells, focusing on the detection of phosphatidylserine and HMGB1 protein to differentiate between apoptotic and necrotic cell death. This technique allowed for the precise determination of drug concentrations effective against breast cancer stem cells, providing valuable insights into their susceptibility to treatment [23]. A formulation for a targeted and pH-sensitive niosomal system (pHSN) was developed, incorporating quantum dot (QD)-labeled Trastuzumab (Trz) molecules.…”

“…Advanced Microscopy and Imaging Techniques: Advanced imaging techniques such as confocal microscopy, electron microscopy, and total internal reflection fluorescence (TIRF) microscopy play a crucial role in the detailed analysis of QDs at the cellular and molecular levels. These methods allow for the in-depth investigation of the interactions, localization, and functions of QDs within biological systems [7,9,12,22,23,32,48,49,57,74,75,85,102].…”

“…Advanced diagnostics and biomarker research are also central to the utilization of quantum dots in breast cancer, focusing on the early detection of mutations, such as those in the BRCA1 gene [18], and the quantitative analysis of cancer-related proteins like IGF1R [19]. Moreover, the use of QDs in glycophenotype analysis [20,21] and the detection of biomarker co-localization [22,23] highlight the role of these nanomaterials in enhancing the specificity and sensitivity of cancer diagnostics.…”