Martin Rebeca San, Das Priyojit, Xue Tianchun, Brown Morgan Rose, Reis Marques Renata Dos, Essington Michael, Gonzalez Adrian, McCord Rachel Patton
bioRxiv. 2023 Mar 23:2023.03.20.533462. doi: 10.1101/2023.03.20.533462.
UNLABELLED: Bone metastasis remains one of the biggest challenges in the treatment of prostate cancer, and other solid tumors such as breast, lung, and colon. Modeling a complex microenvironment in-vitro, such as the bone niche, requires interrogation of cell-cell interactions, specific extracellular matrix proteins and a high calcium environment. Here, we present a fast and cost-effective system in which commercially available, non-adhesive, cell culture vessels are coated with amorphous calcium phosphate (ACP) as a surrogate for bone matrix. We further present modified protocols for subculturing cells, as well as nucleic acid and protein collection in high calcium samples. We find that prostate epithelial cell lines show increased adhesion and proliferation when cultured in these surfaces, as well as independence from androgen starvation. We observe gene expression changes on ACP surfaces in early adenocarcinoma cell lines which may reflect alterations relevant to prostate cancer progression. SUMMARY STATEMENT: To model the role of calcium in the microenvironment of the metastatic bone niche, we developed a cost-effective way to coat cell culture vessels in bioavailable calcium, and show that it has an effect on prostate cancer cell survival.
未标注:骨转移仍然是前列腺癌以及其他实体瘤(如乳腺癌、肺癌和结肠癌)治疗中面临的最大挑战之一。在体外模拟复杂的微环境,如骨龛,需要研究细胞间相互作用、特定的细胞外基质蛋白和高钙环境。在此,我们展示了一种快速且经济高效的系统,其中使用市售的非粘附性细胞培养容器,用无定形磷酸钙(ACP)进行包被,作为骨基质的替代物。我们还展示了用于细胞传代培养以及在高钙样品中收集核酸和蛋白质的改良方案。我们发现,前列腺上皮细胞系在这些表面培养时,粘附和增殖增加,并且不依赖雄激素剥夺。我们观察到早期腺癌细胞系在ACP表面的基因表达变化,这可能反映了与前列腺癌进展相关的改变。 总结陈述:为了模拟钙在转移性骨龛微环境中的作用,我们开发了一种经济高效的方法,用生物可利用的钙包被细胞培养容器,并表明它对前列腺癌细胞的存活有影响。
Acta Biomater. 2019-3-21
Mater Sci Eng C Mater Biol Appl. 2020-11