TY  - JOUR
TI  - A Model of Breast Cancer Micrometastasis in a Three-Dimensional (3D) Liver Spheroid for Testing an Antimetastatic Therapy
AU  - Nevskaya, Kseniya V.
AU  - Pershina, Alexandra G.
AU  - Efimova, Lina V.
AU  - Sukhinina, Ekaterina V.
AU  - Kozlova, Polina K.
AU  - Ryzhkova, Alina Yu.
AU  - Hmelevskaya, Ekaterina S.
AU  - Ibragimova, Marina K.
AU  - Tsydenova, Irina A.
AU  - Litviakov, Nikolai V.
AU  - Udut, Elena V.
VL  - 15
IS  - 18
PY  - 2025
DA  - 2025/09/20
SP  - e5454
C1  - Bio-protocol 2025;15:e5454
DO  - 10.21769/BioProtoc.5454
UR  - https://doi.org/10.21769/BioProtoc.5454
AB  - Even though the survival and proliferation stages of cancer cells that have newly settled at a metastatic site are the rate-limiting stages and the most promising targets for drugs, there is a lack of models of the earliest stage of metastasis formation. A method for modeling breast cancer liver metastasis is described here: a stage of transition of a differentiated tumor cell into a cell actively proliferating in a three-dimensional (3D) liver spheroid. Opposite to existing heterocellular 3D models of metastases, the protocol allows modeling the initial stage of liver colonization by metastatic cells, the so-called “micrometastases.” The method includes obtaining a line of fluorescent tumor cells, fluorescence-activated sorting of differentiated cells, preparing a single-cell suspension of liver cells, forming a liver spheroid in an agarose mold, inducing the tumor cell dedifferentiation and proliferation using IL-6, and intravital microscopy of spheroids, with subsequent processing and analysis of fluorescent images in the ImageJ software. The performance of the proposed model was demonstrated using microRNA therapeutics. The ability of a combination of microRNAs to suppress the transition of micrometastasis to macrometastasis in the 3D liver spheroid was confirmed by an immunofluorescent assay of spheroid sections and transcriptome analysis.
KW  - Spheroid
KW  - Metastasis
KW  - Tumor
KW  - Anticancer drug
KW  - microRNA
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False