Macro- and micro-nutrient–based multiplex stress conditions modulate in vitro tumorigenesis and aggressive behavior of breast cancer spheroids

Abstract

PurposeThe aggressive nature of a tumor is presumably its inherent one, but different environmental cues can manipulate it in many ways. In this context, the influence of metabolic stresses on tumor behavior needs to be analyzed to understand their far-reaching implications on tumor aggression and dormancy. This work investigates different facets of the tumor, such as tumorigenic capacity, tumor phenotype, and migration, under multiple metabolic stress conditions. Methods Non-invasive and invasive multicellular spheroids (MTS) were created and subjected to multiple stress conditions, namely glucose, amino acid, and oxygen deprivation. Altered behavior of the MTS has been evaluated in the context of in vitro tumorigenesis, spheroid formation capacity, phenotype, mRNA profile, migration, and recruitment of mesenchymal stem cells. ResultsThe metabolic stress conditions were observed to negatively impact the in vitro tumorigenesis and spheroid formation process of invasive and non-invasive breast cancer cells. While the stress seemingly influences the growth and phenotype of spheroids, it does not alter the organization of sub-cellular entities significantly. Metabolic stress conditions impact the transcriptomic landscape of hypoxic, angiogenic, ECM deformation, glycolysis shift, and protein starvation-related gene clusters. MTSs do not adhere or migrate under stress, but they exhibit different modalities of migration when rescued. Invasive spheroids, after the rescue, exhibit increased aggressiveness. Furthermore, stressed spheroid was observed to control the migration and recruitment of mesenchymal stem cells. Conclusion Multiplex metabolic stresses could control the tumorigenesis while influencing the physiology of invasive and non-invasive breast cancer spheroids along with their migration pattern and tumor-stromal crosstalk.

The role of decellularized cell derived extracellular matrix in the establishment and culture of in vitro breast cancer tumor model

“…In this presented study, CD-dECM doped organoids were evaluated as 'loose aggregate' in terms of compactness as well as 'irregular with minor concave contour' in terms of circularity, while those without CD-dECM were evaluated as 'tight aggregate-fully remodeled outline' and 'circular' in terms of compactness and circularity, respectively. Although organoids produced entirely from cells, not doped with CD-dECM, seem to be superior to others in terms of compactness and circularity, many studies in the literature have reported the negative effects of collagen and ECM-like molecules doped into the hanging droplet on organoid morphology, especially when used at increasing concentrations [49][50][51][52]. Nevertheless, when assessed beyond morphological criteria, these structures were classified as organoids due to their enrichment with their own ECM and the irregular polarization and morphology of these organoids, resulting from their high protein content in the ECM, were found consistent with the previous literature findings [53,54].…”

Impact of Chemotherapeutic Stress Depends on The Nature of Breast Cancer Spheroid and Induce Behavioral Plasticity to Resistant Population

“…Migration Assay for Control and Treated Spheroids: Pseudo 3D migration and sprouting assay were performed for homotypic and heterotypic spheroids as described previously to analyze the migration capacity of treated spheroids. [87,90] Pseudo 3D migration: Control and drug-treated spheroids were allowed to migrate on a 2D platform (plastic coverslip) filled with fresh media. Migration profile was monitored over 48 h, and images were captured through MagnUS (Olympus) microscope.…”

The Potential of Kaliandra Honey (Calliandra sp.) on Sertoli Cell Counts in Malnourished Albino Rats

“…Defisiensi nutrisi makro dan mikro akibat malnutrisi dapat menginduksi respon stres yang merupakan bagian dari mekanisme pertahanan tubuh (Gayan et al, 2021) Menurut Yao et al, (2001) karakteristik utama apoptosis ditunjukkan melalui kerja protease dan nuklease serta disfungsi mitokondria di dalam sel dengan membran plasma utuh. Apoptosis juga ditunjukkan melalui ciri lain seperti fragmentasi DNA, tonjolan pada membran, penyusutan ukuran sel, dan dekomposisi komponen seluler.…”