Biopolym. Cell. 2011; 27(4):285-290.
http://dx.doi.org/10.7124/bc.00010B
Structure and Function of Biopolymers
Characterization of new cell line stably expressing CHI3L1 oncogene
1Balynska O. V., 2Baklaushev V. P., 1Areshkov P. O., 1Avdieiev S. S., 1Boyko O. I., 2Chekhonin V. P., 1Kavsan V. M.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. V. P. Serbsky National Research Centre for Social and Forensic Psychiatry, RUSA Ministry of Health
    23, Kropotkinsky lane, Moscow, Russian Federation, 119991

Abstract

Aim. To characterize the immortalized 293 cell line after stable transfection with human oncogene (CHI3L1). Methods. 293 cells, stably transfected with pcDNA3.1_CHI3L1, and 293 cells, stably transfected with pcDNA3.1 as a negative control, were used throughout all experiments. The clones of CHI3L1-expressing 293 cells and 293 cells, transfected with pcDNA3.1, were analyzed by immunofluorescence and confocal microscopy. Cell proliferation was measured using MTT assay; analyses of ERK1/2 and AKT activation and their cellular localization were performed with anti-phospho-ERK and anti-phospho-AKT antibodies. Specific activation of MAP and PI3 kinases was measured by densitometric analysis of Western-blot signals. Results. The obtained results show quite modest ability of CHI3L1 to stimulate cell growth and reflect rather an improved cellular plating efficiency of the 293 cells stably transfected with pcDNA3.1_CHI3L1 as compared to the 293 cells transfected with an «empty» vector. ERK1/2 and AKT are activated in the 293_CHI3L1 cells. In these cells phosphorylated ERK1/2 were localized in both cell cytoplasm and nuclei while AKT only in cytoplasm. The 293_CHI3L1 cells differed from the 293 cells, transfected with an «empty» vector, in their size and ability to adhere to the culture plates. Conclusions. The overexpression of CHI3L1 is likely to have an important role in tumorigenesis via a mechanism which involves activation of PI3K and ERK1/2 pathways. The tumors which can be induced by orthotopic implantation of the transformed human cells with overexpressed human oncogene CHI3L1 into the rat brain can be used as a target for anticancer drug development.
Keywords: chitinase 3-like 1 protein (CHI3L1), brain tumor, MAP kinase, PI3 kinase
Full text: (PDF, in English)

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