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Information × Registration Number 0216U005007, 0115U006687 , R & D reports Title Production of stable sublines of cancer cells expressing phenotypical and molecular properties of cancer stem cells (CSCs) for development and monitoring of efficiency of novel antitumor pharmaceuticals popup.stage_title Head Drobot L.B., Registration Date 11-01-2016 Organization A.V.Palladin Institute of Biochemistry of National Academy of Sciences popup.description2 The primary aim of the project was to produce of stable subline of breast, lung and colon cancer cells with up- or down-regulated Ruk/CIN85 adapter protein expression, to investigate of the role of the signal protein in attainment of such functional and molecular properties of cancer stem cells (CSCs) as the ability to form mammospheres in determined medium on low-adhesive surface, expresiion of certain surface markers, drug resistance, increased number and activity of membrane ABC transporters, aldehyde dehydrogenase (ALDH) activity, enhaced migratory and invasive properties, to demonstrate the value of the produced sublines for screening and selection of anti-CSCs pharmaceuticals. We used the following methods: cell culture technique, spheroid culture growth, flow cytometry, MTT assay, PAAG protein electrophoresis, Western-blot assay, fluorometric ALDH activity assay, migration and invasion in vitro assay in modified Boyden chamber, real-time polymerase chain reaction (qPCR), mouse in vivo carcinogenesis model, morphological evaluation of tumor samples. It has been demonstrated for the first time, that a stable overexpression of full-length Ruk/CIN85 in cells of human breast adenocarcinoma line MCF-7 and human lung adenocarcinoma line A549 is associated with attainment of functional and molecular properties that are characteristic of cancer stem cells and contribute to increased migration, invasion and metastasizing, and to resistance towards anticancer pharmaceuticals. We used stable sublines (B1 and B3) of mouse Lewis lung carcinoma (LLC) with downregulated expression of Ruk/CIN85 adapter protein through siRNA-interference to evaluate its participation in control over migratory and invasive properties of cancer cells. It has been demonstrated that downregulated Ruk/CIN85 expression in LLC cells is associated with nearly 33% decreased tumor growth after a short latency, decreased primary tumor mass and volume, lower metastasis count and size (by 80 to 85%) in vivo. These results indicate the involvement of Ruk/CIN85 in development of functional and molecular properties of cancer stem cells (CSCs), that are important for tumor progression in vivo. The produced sublines of breast and lung adenocarcinomamay be used for wide screening of anti-CSCs drugs. Product Description popup.authors Комісаренко С.В. Пасічник Ганна Володимирівна Петухов Дмитро Миколайович Шандренко С.Г. popup.nrat_date 2020-04-02 Close